Canine mammary

Question 1

Lamp et al. 2013. The metastatic potential of canine mammary tumors can be assessed by mRNA expression analysis of connective tissue modulators

Relaxin is linked to metastatic breat cancer in women

Intratumoral relaxin mRNA expression and relaxin plasma levels had no prognostic value

High mRNA levels RXFP1 (relaxin receptor) were an independent marker of metastatic potential with 15-fold risk increase and a predictor for shorter survival

MMP-2 was associated with early death bacuse of canine mammary tumor

The mRNA expression of relxin, RXFP1 and MMP-2 were positively correlated suggesting common linkage

RXFP1 is proposed as possibel therapeutic target in CMT

Question 2

Raposo et al. 2014. Prognostic value of tumor-associated macrophages (TAM) in canine mammary tumors

TAMs associated with poor porgnosis in human breast cancer

The TAM value was significantly ____ in malignant than bening CMT

In malignant CMT, TAMS were associated with?

Survival analysis showed?

Answer 2

Higher

Skin ulceration, histologic type, nuclear grade, tubular differentiation

Tumors with high levels of TMAS and decrease OS

Question 3

Pena et al. 2013. Prognostic value of histological grading in noninflammatory canine mammary carcinoma in a prospective study with 2-year follow up: relationship with clinical and histological characteristics

The tumor size, clincial stage, histological diagnosis, presence/absence of myoepithelial proliferation, regional LN mets at diagnosis were associated with histologic grade

Histologic grade, age, clincial stage, tumor subtype, LN mets, were associated with recurrences and/or metastases, cancer associated death, survival times

Subdivision of stage I (T1NOMO) into stage IA and IB was proposed in terms of prognosis

The clinical stage, histological grade, spay staus were selected as independent prognostic variable (multivariable) with DFS as dependent variable

Question 4

Sanchez-Cespedes et al. 2013. Use of CD10 as a marker of canine mammay myoepithelial cells

5 different cell types were identified

Type 1 cells (fusiform cells with ME cell phenotype - calponin - and CK14+, CK8/18-) were normal cells and found in all samples

Type 2 cells were normal or neoplastic luminal epithelial cells (calponin-, CK14-, CD8/18-)

Type 3 (type 1 phenotype with variable morphology) and 4 (atypical neoplastic cells with mixed ME/LE phenotype) were restricted to tumors and malignant tumors

Type 5 were found in all sample types (fusiform cells with stromal phenotype)

CD10 antigen is sensitive but not specific marker of ME cells in normal, dysplastic, neoplastic mammary tissue

Question 5

Im et al. 2013. Breed-related differences in altered BRCA1 expression, phenotype and subtype in malignant canine mammary tumors

139 malingnant CMT from 5 breeds with highest prevalence in Korea

The ___ breed had then highest proportion of malingnat CMT and stong expression of BRCA1

Cytoplasmic and membranous expression of BRCA1 was associated with

Answer 5

Shih-Tzu

ER-, PR- and triple negative phenotype and basal like subtype

Question 6

Valencakova-Agyagosova et al. 2014. Determination of carcinoembryonic antigen and cancer antigen (CA15.3) in bitches with tumors on mammary gland: preliminary report

Levels of CEA in tumor group were high compared to healthy bitches

Levels of CA15-2 was high in tumor group compared to healthy bitches

Question 7

Multiple RT-PCR markers for the detection of circulating tumor cells of metastatic mammary tumors

Question 8

Kim et al. Identification of triple negative and basal like canine mammary carcinomas using basal markers

Determine whether CMC include triple negative and basal like phenotypes by IHC - ER, PR, HER2, 4 basal markers (CK14, CK5/6, p63, and EGFR)

241 CMC, 45 triple negative tumors (ER-, PR-, HER2-) and was assoicated with poor prognosis

In these tumors the expression of CK14, CK5/6, and EGFR was related to clinicopathologic parameters, while expression of p63 was not relevant

Question 9

Sleeckx et al. 2013. Evaluation of immunohistochemical markers of lymphatic and blood vessels in canine mammary tumors

Develop IHC protocol to identify blood and lymphatic vessels in CMT?

Which were the most suitable markers?

Answer 9

Prox-1 (markers of human lymphatic vessel) and CD31 (marker of BV)

Question 10

Yoshida et al. 2013. TGF-B transiently induces vimentin expression and invasive capacity in a canine mammary gland tumor cell line

Investigate the role of TGF-B in CMGT

Treatment of CMGT cell line with TGF-B1 leads to?

Answer 10

transient induction of vimentin (mesenchymal marker)

Invasivness is increased but reversed with prolonged stimulation

Question 11

Santos et al. 2013. Identification of prognostic factors in canine mammary malignant tumors: a mutivariable survival study

Determine value of several IHC molecules such as MMP-9 and uPA in stromal cells and Ki-67, TIMP-2 and VEGF in cancer cells

MMP-9 and Ki67 are independent prognostic markers of canine malignant tumors

High stromal expression of uPA and MMP-9 is an aggressive tumors suggest that these are potential targets in post-operative tx in canine mammary tumor

Question 12

Chen et al. 2013. Expression of MAGE-A restricted to testis and ovary or to various cancers in dogs

Positive immunoreactivity in 75% of melanomas including oral, cutaneous, eyelid, and interdigital melanoma in 68.7% of oral and nasal tumors

52. 5% discrete round cell tumots
53. 5% STS

Oral SCC, multicentric lymphoma, and extraosseous OSA showed no expression

Overexpression in melanoma981%), malignant nasal tumors (100%), and TVT (100%)

MAGE-A - indicator of malignancy but not specific for any tumor type

Question 13

Clemente et al. 2013. Different role of COX-2 and angiogenesis in canine inflammatory and non-inflammatory mammary cancer

IHC of angiogenesis markers (COX-2, VEGFA, VEGFD, VEGFR-3, MVD, lymphatic proliferation index (LP), Ki-67) in 21 canine IMC, 20 high grade malignant non-IMC mammary tumors (MMT), and 4 normal samples

The expression of ____, ___, ___, were higher in IMC, MVD, LPI tumors but not proliferation index

MMTs, ___ was asociated with ___ while in IMC, ___ was associated with ___

Lymphagiogenic pathway is specific role of COX-2 in IMC angiogeneis - justifies use of COX-2 therapies

Answer 13

COX-2, VEGF-A, VEGF-D

COX-2 with VEGF-A, COX-2 and VEGF-D

Question 14

Vercelli et al. 2015. Expression and functionality of TRPV1 receptor in human MCF-7 and canine CF.41 cells

TRPV1 is associated with cancer growth and progression

All tested TRPV1 agonists and antagonists caused ___

CF.41 cells capsaicin and capsazepine ___

Answer 14

decrease cell growth in MCF-7 cells

increase in cell proliferation

Question 15

Borge et al. The ESR1 gene is associated with risk of canine mammary tumors

Association to CMT for SNPs and haplotypes in the estrogen receptor 1 (ESR1) gene in ESS material

Large number of SNP in ESR1 showed different allelic frequency betwenn high and low risk breed

CMT-associated SNP suggest this gene might be involved in CMT development

Question 16

Pawlowski et al. 2013. Gene expression profiles in canine mammary carcinoma of various grades of malignancy

Question 18

Kristiansen et al. 2013. Effect of OVH at the time of tumor removal in dogs with benign mammary tumors and hyperplastic lesions: a randomized controlled clincial trial

84 sexullay intact bitches

OHE (42) no OHE (42) at time of NMT (nonmalignant mammary tumor)

New mammary tumors developes in ___ intact dogs and ___ OHE dogs

How many dogs developed new tumors?

Survival between two groups?

OVH reduced the risk of new tumors by 50%

Answer 18

64% and 36%

21%

Not different

Question 19

Ochiai et al. 2013. Molecular cloning and tumor suppressor function analysis of canine REIC/Dkk-3 in mammary gland tumors

REIC/Dkk-3 suppressor of growth in several human cancers

Expression of REIC/Dkk-3 was ___ in mammary gland tumors and in canine mammary carcinoma cell lines than normal mammary gland tissue

Overexpression of REIC/Dkk-3 induced?

Answer 19

Lower

Apoptosis in carcinoma cell lines

Expression is downregulated in canine mammary tumors

Question 20

Pawlowski et al. 2013. Expression and role of PGP, BCRP, MRP1, and MRP3 in multidrug resistance of canine mammary cancer cells

In canine mammary cancer vinblastine efflux is caused by?

Cisplatin efflux mediated by?

Cyclophosphamide resisatnce?

RNA silencing of efflux pumps?

Answer 20

PGP and MRP1 proteins

PGP, BCRP, MRP1, MRP3

BCRP

Decreased IC50 doses of all drugs in mammary carcinoma cells

Treating cell with siRNA targeting efflux pumps could be beneficial

Question 21

Pawlowski et al. 2013. Five markers useful for the distinction of canine mammary malignancy

Gene set - sehrl, zfp37, mipep, relaxin, magi3 - malignancy marker that could help distinguish most malignant canine mammary carcinomas

Question 22

Fukumoto et al. 2013. L-type amino acid transporter (LAT1): a new therapeutic target for canine mammary gland tumor

LAT1 - transports aa’s needed for cellular activities, growth, proliferation

3H-leucine uptake by CHM (cell line) and effects on growth were analysed in presence and absence of LAT1 inhibitors

Uptake and cellular growth in CHM were ___ in dose dependent manner by both LAT1 inhibitors

Inhibitories growth actiites of various chemo drugs were enahcned by combining the LAT1 inhibitors

Answer 22

inhibited

Question 23

Karayannppoulou et al. 2013. Markers of lipid peroxidation and a-tocopherol levels in blood and neoplastic tissue of dogs with malignant mammary gland tumors

Study the extent of lipid peroxidation and cocentration of a-tocopherol inhibitor of lipid peroxidation

16 intact female dogs with malignant MGT and 12 healthy dogs

Difference in BARs, a-tocopherol, total cholesterol, and triglycerides between the two groups?

TBARs and a-tocopherol in malignant tissue compared to non-malignant?

Increased levels of TBARs suggest oxidative stress in canine malignant MGT

Answer 23

No difference in serum levels of TBARs, a-tocopherol, total cholesterol, and triglycerides between the 2 groups

TBARs was higher and a-tocopherol lower in neoplastic tissue when compared with norml mammary gland tissue

Question 24

Camacho et al. 2013. Establishment and characterization of a canine xenograt model of inflammatory mammary carcnima

Model is hormone receptor positive and HER2-

Estrogens and androgens are locally produced in tissues

Factors related to tumor vascularization showed positive expression and xenografts with the highest expression of all analyzed vascular factors had highest rate of tumor proliferation

Question 25

Im et al. 2014 Analysis of a new histological and molecular-based classification of canine mammary neoplasia

Canine mammary neoplasm were analyzed according to grading system proposed by Goldshmidt et al.

Canine mammary neoplasm account for? Of these hoe many were malignant?

What grade were the carcinoma-anaplastic subtype?

What grade were the carcinoma-tubular subtype (83.3%) and complex adenoma/mixd tumor subtype (85.2%)?

What was the most common finding with comedocarcinoma, carcinom-anaplastic, and inflammatory carcinoma subtype?

Most freqeuntly occuring molecular subtype?

Which subtype was associated with grade 3 tumors and lymphatic invasion?

Answer 25

52.6%, 51.7%

Grade 3 (100%)

Grade 1

Tumor cell invasion into lymphatic vesses

Luminal A (44%)

basal-like subtye

Question 26

DeInnocentes et al. 2015. Characterization of HOX gene expression in canine mammary tumor cell lines from spontaneous tumors

Spatial/temporal controls of development are regulated by homeostatic (HOX) gene complex

Determine if HOX expression profiles are associated with neoplasia as they pose in people

Five canine HOX genes were overexpressed with expression profiles consistent with oncogene-like cluster (HOXA1, HOXA13, HOXD4, HOXD9, and SIX1) and 3 HOX genes wee underexpressed (HOXA11, HOXC8, and HOXC9) and nonsense mutation in HOXC6

Question 27

Camacho et al. 2014. Immunohistochemical vascular factor expression in canine inflammatory mammary carcinoma

IMC xenografts showed higher COX-2 expression assoviated with VEGF-D and VEGFR-3 as well as higher presence of dermal lymphatic tumor emboli, suggesting COX-2 participation in IMC lymphagionesis

Question 28

Carvalho et al. 2013. EGFR and microvessel density in canine malignant mammary tumors

61 malignant neoplasms were studied with IHC comparing expression of EGFR and MVD by CD31 labeling

Higher EGFR expression was significantly associated with?

High CD31 was significcantly associated with?

Correlation between EGFR and CD31?

Answer 28

Tumor size, tumor necrosis, mitotic grade, histological grade of malignancy and clinical stage

tubule formation, histologic grade, clinical stage

Positive correlation

Overexpression of EGFR may contribute to increased angiogenesis and aggression in malignant CMT - possibility of using EGFR inhibitors

Question 29

Beck et al. 2013. Genome abberations in canine mammary carcinomas and their detection in cell-free plasma DNA

Sneuploid in 2 tumors, frquent smaller deletions in 1, inter-chromosomal fusion in one, 1 normal tumor

Abberations affect several cancer associated genes such as cMYC and KIT

Once common deletion of the proximal end of CFA27, harboring the tumor suppressor gene PFDN5 was detected in 4 tumors - detected in 50% of tumors

Question 30

Maichrzak et al. 2013. Migrastatin anlogues inhibit canine mammary cancer cell migration and invasion

Investigate 6 migrastatin analogues (MGSTA-1 to 6) on migration and invasion of canine mammry ACA cell line

Which were potent inhibitors of migration and invasion?

Treatment of cells with MGSTA-6 disturbed the binding between?

Answer 30

MGSTA-5 and MGSTA-6

F-actin and fascin1 - increased unbound fascin and reduced colocalization of F-actin and fascin1 in canine caner cells

Actin filaments were not corss linked by fascin and did not generate filopodia structure

Question 31

Raposo et al. 2014. Tumor-associated macrophages are associated with vascular endothelial growth factor expression in canine mammary tumors

TAMs were assoiated with malignant CMT and VEGF positive tumors and also associated with VEGF expression within malignant CMT

Association between TAMs and presence of infiltrative growth, low tubule formation, and LN metastasis

TAMs influence angiogenesis in CMT and may present a therapeutic target

Question 32

Magalhaes et al. 2013. Immunodetection of cells with a CD44+/CD24- phenotype in canine mammary neoplasms

CSCs are detected in canine mammary tumors using CD44+/CD24-

The value at CD44 was positive and CD24 becomes negative was 46.75%

Cells with CD44+/CD24- phenotype were detected in 40/130 samples with advantage of high garde tumors (II and III) and metastases among tubular, papillary, and carcinoma in mixed tumors

In these samples, percentage stained by CD44 and CD24 antibodies was 62.2% and 0%

CD44+/CD24- correlated with grade II and III tumors - poor prognosis

Question 33

Sleeckx et al. 2014. Lymphangiogenesis in canine mammary tumors: a rmorphometric and prognostic study

Analyze selected characteristics of lymphatic vessels in CMTs to evaluate prognostic significance

56 bening CMT, 55 malignant CMT, 13 control samples

The intratumor lymphatic vessels were smaller, less numerous and occupied smaller relative area compared to peritumoral region

No differencs in lymphatic vessel parameters in benign or malignant

Question 34

Sleeckx et al. 2014. Angiogenesis in canine mammary tumors: a morphometric and prognostic study

Blood vessels and proliferating endothelial cells were present in intratumoral and peritumoral regions of bening and malignant tumors

Vessels in PT regions had significantly higher area and perimeter compared with IT regions

Malignant tumors showed significantly more vessels with small total blood vessel area and higher blood endothelial cell proliferation compared with benign tumors and control tissue

In the PT region there significantly higher blood vessel density, blood vessel area, and blood vessel perimeter

Question 35

Krol et al. 2014. Macrophages mediate a switch between canonical and non-canonical Wnt pathways in canine mammary tumors

TAMs mediate a switch between canonnical and non-canonical WNT signaling pathway leading to increased tumor invasion and metastasis

TAMs constitutively secrete WNT inhibitors that decrease tumor proliferation and development but as side effcet induce non-canonical Wnt pathway which leads to metastasis

Question 36

Leonel et al. 2014. Expression of glutathione, glutathione peroxidase and glutathione S-transferase pi in canine mammary tumors

GSH in conjunction with GSH-Px and GST pi play a role in detoxification and biotrasnformation of chemotherapy drugs

What was correlated with absence of tumor ulceration and was present in dogs without metastasis?

Signifcant correlation of survival and increase of GSH

Correlation with GSH-Px and GSH-pi and other variables?

Answer 36

High expression of GSH

None

Question 37

Yoshimura et al. 2015. Cellular source of tenascin-C in canine mammary tumors

Tn-C is an ECM glycoprotein implicated in cancer progression in humans

Accumulation of Tm-C has been recognized in prolifearting myoepithelial cells in complex carcinoma, BM zone in low-grade simple carcinoma, and reactive stroma in high grade simple carcinoma

In complex carcinoma, Tn-C was generated by proliferating myoepithelial cells

Tn-C in low-grade simple carcinomas was also derived from myoepithelial cells

Stromal Tn-C in high grade carcinoma was synthesized by fibroblasts/myofibroblasts

Origin of Tn-C differs between low grade and high grade malignancies

Question 38

Bongiovanni et al. 2015. Survivin and related protein in canine mammary tumors: IHC expression

An increase in nuclear survivin expression compared with healthy mammary glans was observed in CMT - nuclear labeling was related to prescence of necrosis

No relation with histological grade or stage

Question 39

Tran et al. 2016. Surgical treatment of mammary carcinoma in dogs with or without postoperative chemotherapy

94 canine mammary carcinomas treated with surgery (58) or surgery and chemotherapy (36)

On multivariable analysis, predictors of MST were?

Complete surgical margins were associated with MST in dogs with stage 1-2 MCA? and lymphatic invasion?

No statistically significant improvement in MST in dogs with stage 4 of LI treated with adjuvant chemotherapu - 5 dogs with complete surgical margins that received mito and carbo had mean survival of 1139 days

Answer 39

Clinical stage, lymphatic invasion (present 179 days and non 1098 days), ulceration (present 118 days, and non 443 days), surgical margins (incomplete 70 days, and complete 872 days)

incomplete 68 days and complete 1098 days. incomplete 70 days and complete 347 days

Question 40

Shinoda et al. 2014. Significance of ERa, HERR2, and CAV1 expression and molecular subtype classification to canine mammary gland tumor

Degree of positive staining for ERa, HER2, and CAV1 showed significant correlations with the behaivour and prognosis of tumors

Question 41

Milanta et al. 2015. COX-2, mPGES-1, and EP2 receptor immunohistochemical expression in canine and feline malignant tumors

COX-2 positivity was observed in 83% canine and 81% feline mammary carcinomas, mPGES-1 in 75% canine and 66% feline, and EP2 was 89% canine and 54% feline

Frequency of COX-2, EP2 receptor and mPGES-1 expression was significantly higher in carcinomas than in non-neoplastic tissue and adenomas

Support the role of COX-2/PGE2 pathway in pathogenesis of these tumors

Question 42

Gracanin et al. 2014. Ligand-independent canonical Wnt activity in canine mammary tumor cell lines associated with aberrant LEF1 expression

Question 43

Rasotto et al. 2014. The dogs as a natural model for the study of the mammary myoepithelial basal cell lineages and its role in mammary carcinogenesis

Basal-like tumours constitute 2-18% of all human breast cancers (HBCs).

We hypothesized that progenitor cells (CK5(+), CK14(+), p63(+) and VIM(+)) differentiate into terminally-differentiated luminal glandular (CK8/18(+)) and myoepithelial (CALP(+), SMA(+) and VIM(+)) cells via intermediary luminal glandular cells (CK5(+), CK14(+) and CK8/CK18(+)) and intermediary myoepithelial cells (CK5(+), CK14(+), p63(+), SMA(+), CALP(+) and VIM(+)).

Neoplastic myoepithelial cells in canine complex carcinomas had labelling similar to that of terminally-differentiated myoepithelial cells, while those of carcinomas-and-malignant myoepitheliomas with a more aggressive biological behaviour (i.e. higher frequency of vascular/lymph node invasion and visceral metastases and higher risk of tumour-related death) were comparable with intermediary myoepithelial cells and had significantly higher Ki67 expression. The majority of CMCs examined were negative for expression of HER-2. The biphasic appearance of CMCs with involvement of the myoepithelial component in different stages of cell differentiation may help to define the role of myoepithelial cells in the mammary carcinogenetic process and the heterogeneous nature of basal-like HBCs.

Question 44

Santos et al. 2016. VEGFR-2 expression in maligna ttumors of the canine mammary gland: a prospective survival study

VEGFR-2 interacts with VEGF-A to promote tumor angiogenesis

VEGFR-2 expression was associated with VEGF immunoreactivity in cancer cells

VEGFR-2 was expressed by endothelial cells from tumor vasculatur and positively associated wutg stromal MMP-9

Carcinosarcomas exhibited high VEGFR-2 suggesting it may be an activated pathway

VEGF anf VEGFR-2 were independed ot patients OS and DFS

Question 45

Mucha et al. 2014. MDSCs mediate angiogenesis and predispose canine mammary tumor cells for metastasis via IL-28/IL-28R (IFN-g) signaling

Increased activation of L-28/IL-28R (IFN-g) signaling in MDSCs

Highest expression of IL-28 was observed in stage III/IV mammary tumor bearing dogs

IL-28 secreted by MDSC stimulates STAT3 in tumor cells which results in increased angiogenic factors and induction of angiogeneis, EMT and increased migration of tumor cells in vitor

Knockdown of IL-28RA decreased angiogenesis, tumor cell invasion and migration

Question 46

Jagarlamundi et al. 2014. Properties of cellular and serum forms of TK1 in dogs with acute lymphocytic leukemia (ALL) and canine mammary tumors (CMT): implications for TK1 as proliferation biomarkers

Serum TK1 protein were significantly higher in CMT than in healthy dogs

Large inactive fraction of TK1 protein in CMT

sTK1 protein assay can differntiate between benign tumors from healthy more efficiently than sTK1 assay

Question 47

Sardon et al. 2015. Absence of canine pappilomavirus sequences in canine mammary tumors

No PV DNA was found in normal or neoplastic canine mammary tissues - canine PVs are probably not involved in the pathogenesis if canine mammary neoplasia

Question 48

Gamba et al. 2014. ZEB2 and ZEB1 expression in a spontaneous canine model of invvasive micropapillary carcinoma of the mammary gland

IHC showed nuclear and cytoplasmic stainining for ZEB2 and nuclear staining for ZEB1

“in situ” areas had higher positivity for cytoplasmic ZEB2 than invasive areas of invasive micropappilary carcinoma (IMPC)

ZEB1 positiveity was associated with low histolotigcal grade

A shorter OS was observed in IMPCs positive for cytoplasmic ZEB2

Cytoplasmic ZEB2 is an important factor in early stages of malignancy and predicts poor OS for IMPC. ZEB1 downregulation appears to be associated with dedifferentiation process of IMPC

Question 49

Guil-Luna et al. 2014. Progresterone receptor isoform A may regulate the effects of neoadjuvant aglepristone in canine mammary carcinoma

Effects of antiprogrestin aglepristone on cell proliferation and mRNA expression of progresterone isoforms A and B in mammary carcinomas treated with 20 mg/kg aglepristone or vehivle 2x before surgery

Total progesterone receptor and isoform A mRNA expression levels decreased after treatment with aglepristone.

Significant decrease in Ki-67 cells was observed in progesterone. psotive and isoform A positiev tumors in aglepristone-treated dogs

Antiproliferative effects are mediated by progresterone receptor isoform A

Question 50

Tien et al. 2015. Downregulation of the KLF4 TF inhibits the proliferation and migration of canine mammary tumor cells

Kruppel-like factor 4 is a TF associated with proliferation, differentiation, migration, and apoptosis

Identified as an oncogene in human breast cancer

KLF4 is expressed in various normal canint tissues and downregulation of KLF4 inhibited CMT cell proliferation and migration and induced cell death

KLF4 may represent a therapeutic target

Question 51

Lim et al. 2015. Obesity, expression of adipocytokines, and macrophade infiltrastion in canine mammary tumors

The mean age of MC development was lower in overweight dogs (9 yr) than in lean dogs or optimal bodyweight (10 yr)

Evidence of lymphatic invasion was found more frquently in owerweight or obese group than in lean groups

Decreased adiponectin expression and increased macrophade numbers in verweight and obese subjects were significantly correlated with factors related to poor pronosis - high histological grade and lymphatic invasion

Leptin expression was correlated with progesterone receptor status, and and leptin receotir eas correlated with estrogen receptor status of MC, regardless of BCS

Question 52

Delgado et al. 2015. Activation of mammalian target of rapamycin in canine mammary carcinomas: an IHC study

p-mTOR was not expressed in normal canine mammary tissue but cytoplasmic labeling was observed in 78% of canine mammary carcinomas

NO relationship between p-mTOR cytoplasmic expression and histological type or grading of carcinomas, degree of tubulue formationm anisokaryosis, mitotic activity or LN mets

p-mTOR is involved in mammary carcinogenesis in dogs

Question 53

Kristiansen et al. 2017. Tolerability and PK profile of a novel benzene-poly-carboxylix acids complex with cis-diammineplatinum (II) dicloride in dogs with malignant mammary tumors

Half-life was 125 h

MTC of BP-C1 was above 0.46 mg/kg

Significant reduction correlated negatively with increasing dose

No decrease in tumor burden was found

Question 54

de Oliveira et al. 2015. Anti-influenza neuraminidase inhibitor oseltamivir phosphate induces canine mammary cancer

Oseltamivir is a anti-influenca sialidase inhibitor

Sialylation, govered by sialyltransferases and sialidases, are implicated in oncogenesis and progression of BC

Oseltamivir impairs canine mammary cancer sialidase activity altering sialylation pattern of canine mammary tumors and to in vitro and in vivo increased mammary tumor aggressive

Question 55

Lim et al. 2015. Effects of obesity and obesity-related molecules on canine mammary gland tumors

The mean age of CMC onset was lower in overwight or obese group (8.7 yr) than in lean or ideal BW group (10.4)

Proportion of poorly differentiated (grade 3) tumors was significantly higher in overweight or obese female dogs

Aromatase expression was higher in overweight or obese grop adnw as correlated with expression of HRs

Overweight ot obese status might affect development and behaivour of CMCs by tumor-adipocyte interaction and increased HR-related tumor growth

Question 56

Borge et al. 2015. Canine mammary tumors are affected by frequent copy number aberration, including amplification of MYC and loss of PTEN

Results: We found a high occurrence of copy number aberrations in canine mammary tumours, losses being more frequent than gains. Increased frequency of aberrations and loss of heterozygosity were positively correlated with increased malignancy in terms of histopathological diagnosis. One of the most highly recurrently amplified regions harbored the MYC gene. PTEN was located to a frequently lost region and also homozygously deleted in five tumours. Thus, deregulation of these genes due to copy number aberrations appears to be an important event in canine mammary tumour development. Other potential contributors to canine mammary tumour pathogenesis are COL9A3, INPP5A, CYP2E1 and RB1. The present study also shows that a more detailed analysis of chromosomal aberrations associated with histopathological parameters may aid in identifying specific genes associated with canine mammary tumour progression.

Conclusions: The high frequency of copy number aberrations is a prominent feature of canine mammary tumours as seen in other canine and human cancers. Our findings share several features with corresponding studies in human breast tumours and strengthen the dog as a suitable model organism for this disease.

Question 57

Saeki et al. 2015. Phenotypic screening of a library of compounds against metastatic and non-metastatic clones of a canine mammary gland tumor cell line

Question 58

Hennecke et al. 2015. Prevalence of Prefoldin subunit 5 gene deletion in canine mammary tumors

Somatic deletion at proximal end of canine chr 27 (CFA27) ws reported in 50% of mammary tumors - harbors tumor suppressor gene PFDN5

Deletion in 24% of mammary tumors, 7.5% in benign tumors and not present in normal mammary tissue

Solid carcinomas showed the highest frequency of deletion (675) and those malignomas without microscopical high fraction of benign tissue had 32% frquency

Ki-67 score was higher in PFDN5-deleted group compared to malignant tumors without the deletion

Question 59

Shin et al. 2015. Analysis of HIF-1a expression relative to other key factors in malignant canine tumors

HIF-1a expression correlated with histological type, grade, negativity of ER and expression of Ki-67 and VEGF

Lymphatic invsion, molecular subtype, PR, HER2 and E-caherin levels did not correlated with HIF-1a expression

Question 60

Im et al. 2015. CD44+/CD24- cancer stem cells are associated with higher grade of canine mammary carcinomas

Single markers, both CD44+ and CD24+ were associated with less aggressive histological subtypes, low grade, and non-TN subtype. Both markers were associated with HR positivity

CD44+/CD24- was associated with higher grade or carcinoma

Question 61

Beirao et al. 2015. Canine mammary cancer cells direct macrophaddes toward an intermediate activation state between M1/M2

TAMs express an activated phenotype, M2, which sustain proliferation of cancer cells, and associated with poor clincial outcome in human cancer patients

Cancer cells inhibit LPS-induced macrophage activation

Macrophage associated proteins, CSF-1, and C-C motif ligant (CCL-2) stimulate macrophages and responsible for effects of cancer cells on macrophages

Suggest feedback loop between marcophages and cancer cells, while cancer cells influence the phenotype of TMAs through CSF-1 and CCL2, macrophages induce canine mammary cells to upregulate their owen expression of the receptors for CSF-1 and CCL2 and increase the cancer cellular metabolic activity.

Question 62

Yoshikawa et al. 2015. Reduced canine BRCA-2 levels in mammary gland tumors

Expression of canine BRCA-2 in mammary tumor samples was lower than in mammary gland samples

No mutations in the canine BRCA-2 were found that altered BRCA-2 transcription levels

Question 63

de Oliveira et al. 2015. Hypoxia upregulates Galectin-3 in mammary tumor progression and metastasis

Galectin-3 is a member of b-glactoside-binding family of lectin and has been implicated in metastasis

In malignant CMT cells, hypoxia induced expression of galectin-3

Under hypoxia, expression shifts from nuclear location to cytoplasmic and membrane expression

Tumor hypoxia upregulates galectin-3 which may increase tumor aggressiveness

Question 64

Pavelski et al. 2015. Infrared thermography in dogs with mmary tumors and healthy dogs

Imaging exam used for breast cancer diagnosis in humans

Higher temperature in the caudal abdominal and inguinal mammary glands than the other glands in the healthy group

Dogs with mammary tumors has signifcantly higher thermographic temperature compared with unaffceted glands regardless of size and location

Technique seems to be able to assess the presence of neoplasia within the mammay tissue in bitches

Question 65

Okada et al. 2015. Localization of heat shock protein 110 in canine mammary gland tumors

HSPs are improtant for protein folding, conformation, and asseembly

Previus study HSP110 is a canine mammary gland tumor antigen and mRNA expression is increased in tumor tissue

IHC showed HSP110 was localized in cytoplasm of epithelial and interstital cells in canine MGT

Question 66

Ebisawa et al. 2015. Significance of caveolin-1 and MMP-15 gene expression in canine mammary tumors

Cav-1 and MMP-14 genes were highlly expressed in CMT tissues compared to normal tissues

Cav-1 was especially overexpressed in malignant and invasive CMT tissues

Localization of Cav-1 was observed on invadopodia-mediated degradation sites of gelatin matrix

Cav-1 may be involved in CMT invasion

Question 67

Garcia et al. 2017. Tumor necorosis factor-alpha-induced protein 8 (TNFAIP8) expression associated with cell survival and death in cancer cell lines infected with canine distemper virus

CDV. replication induced cytopathic effect, decreased cell proliferation rate, and >45% of infected cells were considered death or under late apoptosis/necrosis

TNFAIP8 and CDVM gene expression were correlated in all cell lines

Question 68

Gamba et al. 2015. The relationship between E-cadherin and its transcriptional repressors in spontaneously arising canine invasive micropappilary mammary carcinomas

Canine mammary IMPCs had loss of E-cadherin from carcinoma in situ to invasive areas which appears to be induced by transcription factor SNAIL

In LN metastasis, ZEB1 appears to not exert E-cadherin transcriptional repression activity

Question 69

Queiroga et al. 2017. Quantification of EGFR in canin mammart tumors by ELISA assay

Higher tissue EGFR were found in CMT compared with controls

In malignant CMT, tissue EGFR elevated concentrations were significantly assocaited with tumor relapse and/or distant metastasis amnd reduced disease-free and OS

The IMC had highest EGFR compared with other malignant non-IMC tumors

Question 70

Qui et al. 2015. Promoter mutation and reduced expression of BRCA1 in canine mammary tumors

46.7% of malignant mammary tumors were found with deletion of 1 cytosine in the promoter region

The mRNA expression of BRCA1 was significantly reduced in benign and malignant mammary tumors, and protein expression of BRCA1 was significantly reduced in malignant mammamry tumors

Question 71

Pandley et al. 2015. Mammoglobin as a diagnostic serum marker of complex canine mammary carcinomas

Reported to act as a marker for breast cancer for women

90% sensitive and 95% specific for cut-off 0.177

It can act as novel molecular marker in detecting mammary tumors in canine

Question 72

Raposo et al. 2017. Exploring new biomarkers in the tumor microenvironment of canine inflammatory mammary carcinoma

Gene expression differences between CIMC and non-CIMC were obatined for COX-2, synuclein gamma (SNCG), tribbles 1, VEGF, CSF1R

Correlations were found between SNCG and tribbles 1

Question 73

Cardoso et al. 2017. A 3D cell cultre system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators

Question 74

Diab et al. 2017. Production and characterization of monoclonal antibodies for canine CD138 (syndecan 1) for nuclear medicine preclinical trials on spontanoeus tumors

CD138 is frequently expresssed in canine mammary carcinoma corresponding to human TN subtype with cytoplasmic and membranous expression

In canine DLBL, CD138 is assocaited with non-germinal center phenotype corresponding to most aggressive subtype in humans

Question 75

Caceres et al. 2017. Canine cell line, IPC-366, as a good model for the study of inflammatory breast cancer

Inflammatory breast cancer (IBC) is an aggressive type of cancer with poor survival in women. Inflammatory mammary cancer (IMC) in dogs is very similar to human IBC and it has been proposed as a good surrogate model for study the human disease. The aim was to determine if IPC-366 shared characteristics with the IBC cell line SUM149. The comparison was conducted in terms of ability to grow (adherent and nonadherent conditions), stem cell markers expression using flow cytometry, protein production using western blot and tumorigenic capacity. Our results revealed that both are capable of forming long-term mammospheres with a grape-like morphology. Adherent and nonadherent cultures exhibited fast growth in vivo. Stem cell markers expressions showed that IPC-366 and SUM149 in adherent and nonadherent conditions has mesenchymal-like characteristics, E-cadherin and N-cadherin, was higher in adherent than in nonadherent cultures. Therefore, this study determines that both cell lines are similar and IPC-366 is a good model for the human and canine disease.

Question 76

Soultani et al. 2017. Assessment of sentinel LN metastasis in canine mammary gland tumors using CT indirect lymphography

The absence of opacification or heterogenoeus opacification 1 min after contrast showed the highest sensitivity, specificity and accuracy (93%, 100%, 98.4%)

Images taken 1 min after injection, absolute density value lower than 444 Hounsfield units in the center of SLN also provided significant sensivity and specificty (94% and 75%)

The size and shape of LN showed the lowest sensivity and specificity

Question 77

Kau et al. 2017. S100A4 (metastasin) positive mesenchymal canine mammary tumor spheorids reduce Tenascin C synthesis under DMSO exposure

DMSO did not affect proliferation

TNC was reduced under DMSO exporuse for 7 and 14 days, and S100A4 effect was seen after 14 days

Without DMSO, cells expressed TNC and S100A4

Question 78

Beffagna et al. 2017. Circulating cell-free DNA in dogs with mammary tumors: short and long fragments and integrity index

cfDNA has been considered diagnostic/prognostic plasma biomarker in tumor-bearing subjects

cfDNA differed in neoplastic vs. nonneoplastic dogs and allowed distinction between malignant and non-malignant lesions

cfDNA could be diagnostic marker in dogs carrying mammary nodules

Question 79

Ozmen et al. 2017. Somatic SNP of the BRAC2 gene at the fragments encoding RAD51 binding sites of canine mammary tumors

BRCA2 gene contains 8 BRC repeats in exon 11 that bind to RAD51

19 SNPs of exon 11 of BRCA2 in canine mammary tumors were detected

The c.2383A (T1425P) SNP was found to be the most probably disease associated SNP

Question 80

Burrai et al. 2017. A first IHC study of transketolase and transketolase-like 1 expression in canine hyperplastic and nonneoplastic mammary tissue

Transketolase (TKT) and transketolase-like 1 (TKTL1) are involved in the pentose phosphate pathway - an alternative meatbolic pathway for glucose breakdown that could promote cancer

TKT expression was higher in hyperplastic lesions and in both benign and malignant tumors compared to normal tissue

TKTL1 were higher in hyperplastic lesions, simple adenomas and simple carcinomas than normal mammary glands

Expression of PPP enzyme varies along the evolution of canine mammary neoplastic lesions - supports role of metabolic change in development of mammary tumors

Question 81

Altamura et al. 2017. Expression and activation of PDGFB-R, mitogen activated protein/intracellular signal-regulated kinase kianse (MEK) and extracellular signal regulated kinase (ERK) in canine mammary tumors

PDGFBR was expressed and hyperphophorylated in majority of tumor samples and tumor derived cell lines

both MEK and ERK were expressed and activated in cell lines and biopsies

Possible role for TRK inhibitors

Question 82

Gelaleti et al. 2017. Melatonin and IL-25 modulate apoptosis and angiogenesis mediators in metastatic (CF-41) and non-metastatic (CMT-U229) canine mammary tumor cells

Melatonnin has oncostatic actions and IL-25 is active in inflammatory processes that induce apoptosis in tumor cells

Cell treated with melatonin, IL-25, and IL-17B

Treatment with melatonin reduced cell viability, all treatments alone and combined increased caspace 3cleaved protein involved in apoptosis and reduced VEGFA

Question 83

Santos et al. 2017. Malignant canine mammary tumors: Preliminary genomic insights using oligonucleotide comparative genomic hybridization analysis

Genomic inbalances were found in all but 2 tumors. Losses more common than gain

Canine chr 9, 22, 27, 34, X were most frequently affected

Dissimilar oligonucleotide array comparative genomic hybridization ratio profiles were observed in multiple tumors from the same dogs

Suggesting heterogeneity in the tumor

Question 84

Karayannopoulou et al. 2017. Evaluation of blood T-lymphocyte subpopulation involved in host cellular immunity in dogs with mammary cancer

Absolute number of blood lymphocytes in unchanged

CD8+ T cells are significantly suppressed in dogs with mammary cancer or stage II or III compared to age-matched healthy controls

Question 85

Saad et al. 2017. Canine mized mammary tumors as model for human breast cancer with osseus metaplasia

Canine mixed mammary tumours (CMMTs) and human metaplastic breast carcinomas (HMBCs) share several histopathological features and risk factors. In both species, these tumours display epithelial and stromal components. HMBCs are rare malignant tumours, but CMMTs are one of the most common mammary tumours in dogs and are more often benign than malignant. In this study, benign (n = 88) and malignant (n = 13) CMMTs were characterized using specific antibodies against oestrogen receptor, progesterone receptor, human epidermal growth factor receptor 2, cytokeratin 5/6, cytokeratin AE1/AE3, vimentin, Ki67, E-cadherin and p63. Cartilage and bone matrices associated with benign and malignant CMMTs were characterized using specific antibodies against BMP4, Runx2, Sox9 and osteopontin. The current study suggested that CMMTs are of epithelial origin, but display a myoepithelial-like differentiation. The findings suggest key roles for Sox9, Runx2 and BMP4 in chondrogenesis and bone formation in CMMTs. The high expression of osteopontin in CMMTs appears to be unrelated to tumour malignancy.

Question 86

de Oliveira et al. 2017. Invasive micropapillary carcinoma of the mmary gland in humans and canines: Clinicopathologic, immunophenotypical, and survival approaches

IMPC is a breast cancer with proclivity for LN mets

HUmans and canines showed moderate histological grade

Pure subtype and embolid were more frrequent in canines

Most canine and human IMPCs were positive for ER and PR

Mortality was higher in canines (94%) than humans (4%)

Question 87

Harting et al. 2017. Dichloracetate affects proliferation but not apoptosis in canine mammary cell lines

Dichloracetate is a pyruvate dehydrogenase kinase inhibitor. Causes metabolic changes in cancerous glycolysis towards oxidative phosphorylation via indrect activation of pyruvate dehydrogenase in mitochondria

Reduced cell proliferation without inducing apoptosis in all cell lines

Question 88

Baioni et al. 2017. Estimating canine cancer incidence: findings from a population-bases tumor registry in northwestern Italy

Incidence rate was 804 per 100,000 dog-years for malignant tumors and 897 per 100,000 for benign tumors

Higher rate for all cancers in purebred dogs, particular Yorkie and Boxers

Most prevalent was cutanoeus mastocytome and hemangiopericytoma, and mammary gland complex carcinoma and simplex carcinoma

Question 89

Case et al. 2017. Identification of prognostic collagen signatures and potential therapeutic stromal targets in canine mammary gland carcinoma

Collagen density and fiber organization have been shown to regulate tumor progression in mouse and human mammary tumors

Collagen density, fiber width, lenght and straightness were inversely correlated with OS

Grade 3 cases were less likely to have tumor-stromal boundary and lack of boundary predicted poor outcome

Lack of defined-stromal boundary and increased collagen fiber width were associated with decreased survival

Question 90

Waters et al. 2017. Life course analysis of the impact of mammary cancer and pyometra on age-anchored life expentancy in female Rottweilers: Implications for envisioning ovary conservation as a strategy to tpomote healthy living in pet dogs

Mammary carcinoma was diagnosed in 8% females, median age 8.5 yr, case fatality 37%

Pyometra. was diagnosed 6.6%, median age 5.4 yrs, case fatality 7%

Median lifetime of ovary exposure was 4.3 yrs

Risk for developing both diseases increased with longer ovary exposure, it was also associated with an overall longetivity - 33% decrease in mortality, living 17 months longer than females with shorter ovary exposure

Question 91

Cordeiro et al. 2017. Transcriptomic profile reveals molecular events associated to focal adhesion and invasion in canine mammary gland tumor cell lines

Great they developed another cell line

Question 92

Sanchez-Cespedes et al. 2017. Vitamin D receptor expression in canine mammary gland and relationship with clinicopathologic parameters and PR/ER

VDR is ligand TF and mediates action of calcitriol - active product of Vit D synthesis. Actions of calcitriol include capacity to modulate cancer features - proliferation, differentiation, apoptosis, angiogenesis, invasion and metastasis

VDR-positive staining was foound in nuclei of both myopeithelial and luminal epithelial cell layers

VDR expression was higher in nomal mammary tissue (100%) then benign tumors (40%) and malignant tumors (26.5%)

Female dogs >10 yrs had lower VDR expression compared with younger dogs

Question 93

Ryu et al. 2017. Expression of the glutamine metabolism-related protteins glutaminase 1 and glutamate dehydrogenase in canine mammary tumors

Expression of glutaminase 1 (GLS1) in epithelial region increased with grade.

In the stromal region, complex-type tumors displayed signifcantly higher GLS1 intensity than simple-type tumors

glutamate dehydrogenase expression did not have the same results

Question 94

Dolka et al. Retrospective study and immunohistochemical analysis of canine mammary sarcomas

4% of CMT were classified as CMS and represented 5.1% of malignant CMT

Mean age 11 years

Large breed dogs were affected (38.7%)

Majority were FSA (2.1%)

All expressed vimentin and higher levels were noted in FSA and OSA

Ki67 expression was correlated with grade of CMS

Question 95

CSF-1R as an inhibitor of apoptosis and promoter of proliferation, migration and invasion of canine mammary cancer cells

CSF-1R silencing increased apoptosis, decreased proliferation, and decreased migration

Treatment with CSF-1 had opposite effect

CSF-1R may be a therapeutic target.

Question 96

Liu et al. Expression of autophagy-related protein beclin-1 in malignat canine mammary tumors

Cytoplasmic expression of beclin-1 was weaker in cancer cells than normal mammary glands

Low cytoplasmic expression (57%) was associated with older age, lower degree of tubular formation, increased mitotic activity, higher grade, and necrosis

Low nuclear expression (40%) was connected with older age, lower degree of tubular formation, necorsis, and negative Her2/neu overexpression

Univariate analysis - Beclin-1 cytoplasmic expression was poor predictor for overall survival

Multivariate analysis - beclin-1 cytoplasmic expression is independent prognostic factor

Question 97

Prevalence of glomerulonephropathies in canine mammary carcinoma

Renal biopsies from 32 bitches with spontaneous mammary gland adenocarcinoma and 11 control dogs without mammary neoplasia

Light microscopy abnormalities were found in 78% of dogs with mammary carcinoma and non in control group

Focal glomerular mesangial matrix expansion.was the most common alteration (60%), but mesangial cell proliferation (36%) and focal segmental glomeryloschlerosis (24%) were also frequent in dogs with neoplasia

Immunoflorescence staining revealed strong IgM staining (64%) in carcinoma dogs

TEM revealed faint sun-endothelil and mesangial deposits of electron dense material (78%). Mesangial cell interpositioning and segmental effacement of podocyte foot proceses were identified in some (45%)

Changes in glomerulus and proteinuria are common in dogs with mammary carcinoma

Question 98

Clinical staging in bitches with mammary tumors: Influence of type and histological grade

Question 99

Absence of significant AE following thalidomide administration in bitches diagnosed with mammary gland carcinoms

29 dogs tx with high dose og 20 mg/kg/day for 3 months then low dose 10 mg/kg/day for 3 months

Clinical AEs were absent in 55% after HD

An initial 3-5 day period of somnolence was described (14%), prolonged somnolence (17%) and short lasting few hours (10%) and difficult to rouse (17%)

With dose reduction, AE improved in all patients

Within the reference range, erythrocytes, Hct, total leukocyte count, neutrophils, lymphoycytes, monocytes, and GG showed signifcant alteration with thalidomide treatment

Question 100

Immunohistochemical expression of PGP and breast cancer resistance in canine mammary hyperplasia, neoplasia and supporting stroma

Samples included 47 hyperplastic tissues and 10 benign and 46 malignant neoplasms.

PGP and BCRP expression showed that both markers are potentially expressed by epithelial cells, myoepithelial cells in complex tumours and mesenchymal cells in mixed tumours, but expression of both proteins was significantly higher in malignant epithelial cells versus hyperplastic epithelium or the epithelium of benign tumours.

BCRP showed significantly higher expression in epithelial cells of simple carcinomas versus those of complex and mixed carcinomas.

Grade II and III carcinomas had higher epithelial PGP expression than grade I tumours. The positivity of stromal fibroblasts was higher in histological stage II versus I carcinomas, and in histological grade II versus I carcinomas.

Malignant and invasive tumours were more likely to express PGP and/or BCRP in luminal and stromal components and evaluation of these markers could provide valuable information for the identification of tumours characterized by an aggressive and chemoresistant phenotype.

Question 101

Significance of EXH2 expression in canine mammary tumors

EZH2 is one of the catalytic subunits of polycomb repressive complex 2, a epigentic regulator

EZH2 protein was overexpresssed in canine mammary carcinoma

IHC expression was associated with degree of malignancy in canine mammary carcinoma

Question 102

Intratumoral Foxp3 expression is associated with angiogenesis and prognosis in malignant canine mammary tumors

80 malignant CMT

Abundant Foxp3Treg were associated with tumor necrosis, high mitotic rate, marked nuclear polymorphism, poor differntiation, high grade, presence of neoplastic emboli, and LN mets

Intratumoral FoxP3 was correlated with MVD and VEGF

Tumors with abundant FoxP3Treg were associated with shoter OS

Question 103

Her-2 and EGFR mRNA expression and its relationship with versican in malignant-producing tumors of the canine mammary gland

Versican expression promoted tumor growth by destabilizing focal cell contacts, impeding cell adhesion and migration

EGFR mRNA expression showed significant difference between in situ and invsive carcinomatous areas in low and high versicn expression

Identical results with Her2 mRNA

Tumors with low versican expression showed greater EGFR immunostaining in the in situ areas than in invasive areas

High versican expression had greater EGFR and HER2 staining in in situ areas

Significant EGFR and Her-2 mRNA and protein expression in in situ carcinomatour sites relative to invasive areas suggest these may play a role in tumor progression

Question 104

Her-2, EGFR, COX2, Ki67 expression in LN metastasis of canine mammary carcinoma: association with clinicopathological parameters and overall survival

54 primary mammary carcinomas with LN mets (T1,2,3,N1M0), 29 without mets (T1,2,3,N0M0, 25 canine LN mets

Concordance between the expression of Her2, COX2, and Ki67 and a discordance between EGFR expression in primary mammary carcinomas and paired LN metastasis

Higher Ki67 (>24%), larger tumor size and presence of angiolymphatic invasion in canine primary carcinoma wtih LN mets plus presence of extracapsular extension in LNS mets were related to worse prognosis and shorter OS

Primary mammary carcinomas with high expression of her2, cox2 and ki67 also have high expression in LN mets - expression in LNs was not associated with prognosis

Question 105

Carvalho et al. Ki-67 and PCNA expression in canine mammary tumors and adjacent nonneoplastic mammary glands: prognostic impact by a multivariate survival analysis

A positive and statistically significant correlation between the PI of Ki-67 and PCNA in tumors and adjacent nonneoplastic mammary glands was observed in benign and malignant tumors. Tumor size, skin ulceration, histological type, mitotic index, nuclear grade, differentiation grade, histological grade of malignancy, lymph node metastasis, Ki-67, and PCNA expression in tumors and adjacent nonneoplastic mammary glands were statistically associated with overall survival by univariate analysis in malignant cases (n = 43).

Histological grade of malignancy and high intratumoral PCNA retained their significance by multivariate analysis arising as independent predictors of overall survival. Interestingly, the PI of Ki-67 and PCNA of adjacent nontumoral mammary glands were associated with clinicopathological features of tumor aggressiveness and shorter overall survival, demonstrating the need to better explore this adjacent non-neoplastic tissue.

Question 106

Mucha et al. Immunosuppression in dogs during mammary cancer development

Tregs were higher in tumor bearing dogs than in healthy dogs

MDSCs and p-STAT3 expression was higher in cancer patients than control dog

Canine mammary carcinoma with metas to either LN or internal organs had higher MDSC and Trg than benign mammary tumors or malignant mammary tumors without mets

Changes were noted in the blood (50) and tumor tissue (100) of patients during cancer mammary tumor development

Expression of p-STAT3 and VEGF-C was highest in tumors with metastases

Question 107

Raposo-Ferreira et al. Downregulation of ATM gene and protein expression in canine mammary tumors

ATM gene encodes a protein associated with DNA damage repair and maintenance of genomic integrity

Lower ATM transcripts were seen in benign mammary tumors and carcinomas compared with normal mammary gland

Low ATM protein expression was seen in benign tumors, nonmetastatic carcinomas and primary site of metastatic carcinoma compared with normal mammary gland

No significant difference was seen in ATM gene or protein levels among benign and nonmetastatic and metastatic carcinoma

Level of ATM gene expression or protein expression were not associated with clinical or pathological features or with survival

Question 108

Dolka et al. Evaluation of apoptosis-associated protein (bcl2, bax, cleaved caspace 3 and p53) expression in canine mammary tumors

Bcl2 expression in benign lesions and patient with low TNM stage

Bax, CC3, p53 in malignant CMT

Expression of apoptosis associated proteins was not associated with OS

Positive-p53 status was relatd with poor tumor differentiation, higher MI, invasive growth, necrosis, and occured in larger breed dogs

In the shorter-survival group of dogs (≤18 months), a positive correlation was found between CC3 and Bcl-2 expression; CC3 and MI, ERα and p53 expression, while in the longer-survival group (>18 months) CC3 expression was negatively correlated with ERα, whereas p53 expression was positively correlated with MI.

Question 109

Tong et al. Serum starvation and thymidine double blocking achieved efficient cell cycle synchronization and altered the expression of p27, p53, bcl2 in canine breast cancer cells

In this study, we determined the optimum conditions of serum starvation and TdR and their effects on cell cycle synchronization. We further explored the involvement of PI3K/Akt signaling pathway in the cell cycle synchronization by investigating the expression of three key genes (p27, p53 and bcl-2).

Serum starvation resulted in a reversible cell cycle arrest and synchronously progress through G0/G1. The highest percentage of CHMm cells (87.47%) in G0/G1 stage was obtained after 42 h incubation with 0.5% fetal bovine serum (FBS). TdR double blocking could arrest 98.9% of CHMm cells in G1/S phase (0 h of release), and could arrest 93.74% of CHMm cells in S phase after 4h of release. We also found that the p27, p53, bcl-2 genes were most highly expressed in G0/G1 phase. Our current work revealed that serum starvation and TdR methods could achieve sufficient synchronization of CHMm cells. Moreover, the expression of p27, p53 and bcl-2 genes was related to cyclical movements and apoptosis. Our results will provide a new insight into cell cycle regulation and reprogramming of canine cancer cells induced by serum starvation and TdR blocking.

Question 110

Role of tissue factor expression in thrombin generation by canine tumor cells

CMT25 (high TF-expressing mammary cell line) generated more thrombin than did HMPOS (low TF expressiong OSA cell line)

Thrombin generation was dependent onfactor VII and phophatidylserine and was independent of contact activation.

Thrombin geneation of HMPOS was due to contact activation

Question 111

Effect of melatonin in epithelial mesenchymal transition markers and invasive properties of breast cancer stem cells and human cell lines

Melatonin has an inhibitory role in viability and invasiveness of breast cancer spheres as weall as modulating expression of proteins related to EMT in breast CSCs, suggesting its porteintial as ani-metastatic role

Question 112

Qui et al. Roles of DNA mutation in the coding region and DNA methylation in the 5’ flanking region of BRCA1 in canine mammary tumors

2 point mutations in the coding region of canine BRCA1 in 1 benign mammary tumor sample (4702G>T) and 4765G> T) and in 1 malignant canine mammary tumor sample (3619A>G and 4006G>A).

No mutation in normal canine mammary tissue

4702G>T - terminated translation

4765G> T - replaced glutamate residue with glutamine

3619A>G - threonine to alanine

4006G>A - valine to isoleucine

Question 113

Saba et al. 2016. A comparative oncology study of iniparib defines its PK profile and biological activity in a naturally occuring canine cancer model

DLT - fever, anorexia, diarrhea, neutropenia dn thrombocytpenia - most effects due to carboplatin

MTD of iniparib was not identified

Relevant concentration os the parent drug and metabolites were not found in canine tumor tissues, so unlikely to get clinical response in dogs or humans

Question 114

Synergistic growth inhibitory effect of deracoxib with doxorubicin against a canine mammary tumor cell line, CMT-U27

Question 115

Kristiansen et al. Effect of OVH at the time of tumor removal in dogs with mammary carcinomas: a randomized controlled trial

If OHE at time of mastectomy improves prognosis in dogs

OVH did not decrease hazard of relapse or all cause death in univariable analysis

In multivariable analysis, OVH did not influence hazard of relapse but an interaction was found between ER status and E2 (estradiol)

Decreased hazard of relapse in the OVH group compared to non-OVH in dogs with increased E2

Dogs with grade 2, ER positive tumors or increased pre-surgical E2 likelty benefit from OVH

Question 116

Coleto et al. 2018. Prognostic value of occult isolated tumor cells within regional LNs of dogs with malignant mammary tumors

Clinical signifiance of isolated tumor cells (ITC) within LNs is undefined

35.2% of occult ITC and 2.8% of hidden micrometastasis (MIC) were detected with IHC

ITCs frquently seen in medullay region (60%) and metastasis in corticol region (44%)

No difference in OS in dogs with ITC and without ITC

Question 117

Rico et al. 2018. Expression of the Hippo signaling effectors YAP and TAZ in canine mammary gland hyperplasia and malignant transformation of mammary tumors

Significant increase in TAZ (not YAP) in lobular hyperplasia relative to normal gland - may play a role in epithelial proliferation

Nuclear expression of YAP and TAZ were higher in malignant tumors than benign onw - Hippo dysregulation could play a role in CMT transformation

No differences between grades of tumors

Question 118

Monteiro et al. 2018. Tumor associated macrophages: Relation with progression and invasiveness, and assessment of M1/M2 marcophages in canine mammary tumors

TAMs in CMTs excised from 82 female were qualified as low (<50) or high (>50)

Higher TAMs were associated with clinical stage, tumor type, tumor size, vascular invasion, LN mets, high proliferation rates, vascular microdensity, invasive tumor profiles, and worse prognosis

All macrophages infiltrating malignant tumors with high TAM cound expressed CD206 while benign tumors were infiltrated by macrophages expressing NOS2 - suggeting shift from M1 (activated mac) to M2 (subpopulation in malignant tumors)

TAMs are associated with more aggressive types of mammary cancer in dogs

Question 119

Zambrano-Estrada et al. 2018. Molecular iodine/doxorubicin neoadjuvant treatment impair invasive capacity and attenutate side effect in canine mammary cancer

Molecular iodine (I2) exerts antineoplastic effects on different cancer cells activating re-differentiation pathways.

I2 (10 mg/day) in two administration schemes of Doxorubicin (DOX; 30 mg/m2) in 27 canine patients with cancer of the mammary gland. The standard scheme (sDOX) includes four cycles of DOX administered intravenously for 20 min every 21 days, while the modified scheme (mDOX) consists of more frequent chemotherapy (four cycles every 15 days) with slow infusion (60 min). In both schemes, I2 or placebo (colored water) was supplemented daily throughout the treatment.

Answer 119

mDOX attenuated the severity of adverse events (VCOG-CTCAE) in comparison with the sDOX group.

The overall tumor response rate \for all dogs was 18% (interval of reduction 48-125%), and no significant difference was found between groups.

I2 supplementation enhances the antineoplastic effect in mDOX, exhibiting a significant decrease in the tumor epithelial fraction, diminished expression of chemoresistance (MDR1 and Survivin) and invasion (uPA) markers and enhanced expression of the differentiation factor known as peroxisome proliferator-activated receptors type gamma (PPARγ).

Significant tumor lymphocytic infiltration was also observed in both I2-supplemented groups. The ten-month survival analysis showed that the entire I2 supplementation (before and after surgery) induced 67-73% of disease-free survival, whereas supplementation in the last period (only after surgery) produced 50% in both schemes.

Question 120

Fish et al. Malignant canine mammary epithelial cells shed exosomes containing differentially expressed microRNAs that regulate oncogenic networks

CMEC and CMT cells shed exosomes in vitro that contain differntially expresed miRs. CMT exosomal RNA expresses limited number of miRs that upregulate relative to CMEC and these are predicted to target HR and oncogenic pathways

Question 121

Ledet et al. 2018. BB-Cl-Amidine as a novel therapeutic for canine and feline mammary cancer and feline mammary cancer via activation of the endoplasmic reticulum stress pathway

Inhibiting peptidyl arginine deaminase enzyme (PAD) may be a breast cancer therapy.

Canine and feline cell lines were treated with BB-CLA

BB-CLA reduced viability and tumorigenicty of canine and feline cell lines

Activates ER stress pathways downegulating GRP78 a possible target

Question 122

Pandey et al. 2018. Overexpression of mammoglobin-B in canine mammary tumors

Mammglobin is a breast cancer associated protein and function is not known.

High levels of mammoglobin-B mRNA in CMT compared to controls

Mammoglobin-B protein was overexpressed in 7% of healthy mammary glands and 77% in CMT

No diference with grade of CMT histotypes was observed

Question 123

Chen et al. 2018. Expression and prognostic value of c-met in canine mammary tumors

Positive c-met expression in cytoplasm of mammary epithelial cells at variable levels, and in malignant CMT higher c-met found in carcinomas whose grade, stage, and MI were low and absent metastasis

MST shorter in dogs with malignant CMT with diameter of >5cm, regional LN or distant met, high histologic grade

2-year survival rate was higher in dogs with malignant CMT of higher m-met expression than those with low c-met expression (80% vs. 57%)

Question 124

Canadas et al. 2018. Catechol-o-methyltransferase genotypes are associated with progression and biological behaivour of canine mammary tumors

Question 125

Rossi et al. 2018. The impact of toceranib, piroxicam, thalidomide with or without hypofractionated RT on clinical outcome in dogs with inflammatory mammary carcinoma

14 received medical therapy, 4 got RT and medical therapy

Median TTP?

MST?

Dogs treated with medical therapy ORR? CB? Median TTP and MST?

Dogs receiving medical and RT CB? median TTP and MST?

Answer 125

34 days

109 days

21% and 64%, 28 days and 59 days

100%, 156 days and 180 days

Question 126

Sammarco et al. 2018. Preliminary investigation of extracellular vesicles in mammary cancer of dogs and cats: Identification and characterization

EV are membrane bound vsicles produced by cells and play role in cell to cell communication. Horizontal transfer of bioactive molecules within the tumor microevironment

This study identified and characterized canine and feline mammary tumor cell derived EVs

Question 127

Xavier et al. ZEB1 and ZEB2 transcriptions factors are potential therapeutic targets of canine mammary cancer cells

Progression of cancer can be due to TF associated with EMT - ZEB1, ZEB2, SNAI1, SLUG, STAT3, CDH1

2 cell lines had epithelial like morphology

3 had mesenchymal like morphology

The mesnchymal-like cells are more malignant than epithelial like cells - higher ZEB1 and ZEB2 and lower CDH1 gene expression

Positive correlation between ZEB and tumorsphere number and zie

Question 128

Maues et al. 2018. Common germine haplotypes and genotypes identified in BRAC2 exon 11 of dogs with mammary tumors and histopathological analyses

Investigate the frequency of variants in BRAC2 exon 11 in 48 blood and tissue DNA samples

7 SNPs were found, 3 were evaluated

Im all 22 tumors (except 1) which were clinical stage 2 carried atleast 1 mutant alelle of the 3 variants

A total of 98% of bitches had 1 to 3 polymorphisms of the 7 identifed

Question 129

Caceres et al. 2018. In vitro and in vio effect of flutamide on steroid hormone secretion in canine and human inflammatory breast cancer cell lines

In vivo flutamide reduced tumor size by 55% and 65% and metastasis rates decreased

Androgen levels increased and estrogen levels decreased

Flutamide treatment inhibits cell proliferation and promotes tumor reduction by increasing androgen levels

Question 130

Paczuska et al. 2018. Effectiveness of CO2 laser in an experimental mammary gland adenocarcinoma model

Tumors excised from 48 mice using laser and 48 through scalpel

There was no difference between recurrence rate, metastases, survival time in groups excised with scalpel and CO2 laser

Question 131

Muscatello et al. 2018. Her2 amplification status in feline mammary carcinoma: a tissue microarray flourescence in situ hybridization study

Her2 gene is amplified in subset of feline carcinoma despite the HER2 positive or equivoval protein expression

Question 132

Canadas et al. 2018. Canine mammary tymors: comparison of classification and grading methods in a survival study

2 classification systems: WHO and 2011) and 2 grading based on the human Notingham grade

2 grading systems were applied to 134 female dogs with CMTs

Benign (53%) and malignant (50%) was similar

The 2011 classification divided malignant tumors un more categories those classified as complez, solid, tubulipappilary by WHO

Hitopathological sybtype was associated with survival in both systems

Carcinomas arising in benign tumors, complex carcinomas, and mized carcinoma were associated with better prognosis

Carcinosarcomas and comedocarcinomas had high risk or tumor related death

Univariable analysis - grade was related to survival

Multivariable - age, complete excision, 2 index formulas adapted from human breast cancer studies )tumor size, grade, vascular/lymphatic invasion) were independent prognostic factors

Question 133

Raposo-Ferreira et al. 2018. Characteristics of the EMT in primary and paired metastatic canine mammary carcinomas

Primary tumors had more E-cadherin+/vimentin+ co-expression than their paired LN or distant metastasis

The percentage of E-cadherin+/vimentin+ cells in grade 2 and III was higher than grade 1 tumors

Primary tumors had higher SNAIL/SLUG compared to distant metastasis

Inverse correlation between SNAIL/SLUG and percentage of E-cad+/vim+ cells in primary tumors

Question 134

Brandao et al. DNA methylation status of the ERa gene in canine mammary tumors

Investigate the role ESR1 CpG island methylation (CGI) in ERa expression in canine mammary tumors

6/9 malignant tumors didnot have ERa expression (score 0) and 2 had score 2 and 1 score 4

Lower ERa and ESR1 mRNA were found in malignant mammary tumors than in benign or normal gland

Canine ESR1 had an intragenic and non-promoter associated CGI - different from humans

ESR1 is not regulated by methylation unlike in humans

Question 135

Seung et al. 2018. CD204-Expressing tumor-associated macrophages are associated with malignant, high grade and hormone receptor negative canine mammary gland tumors

CD204, an M2 polarized macrophage receptorwas investigated in 101 cases of CMT

Mean number of CD204+ macrophages were higher in malignant CMT than benign CMT

Were higher in grade 3 than in grade 1 or 2

Higher in HR- malignant malignant CMT than HR+ CMT, and higher in those with lymphatic invsion compared to without lymphatic invasion

Question 136

Miyazaki et al. 2018. Use of skin strechtes for single-stage bilateral mastectomies in a dog and a cat

Wound closure was achieved after single stage bilateral mastectomy was achieved without tension or major complicaiton in both animals

Question 137

Accuracy of four ultrasonography techniques in predicting histopathological classification of canine mammary carcinomas

Question 138

de Souza et al. 2018. Relationship between the inflammatory tumor microenvironment and different histologic types of canine mammary tumors

Association between densities of inflammatory cells, tumor fibrosis and histologic types revealed difference in plasma cells, neutrophils, macrophages, and tumor fibosis.

Suggest associotion between higher number of neutrophils and aggressive mammary tumorsm between high densities of plasma cells, macrophages and CD8+ cells and between low number of profile cells of CD4+ cells and less aggressive tumors

Larger areas of tumor fibrosis showed relation to more aggressive mammary tumors

Question 139

Arbe et al. 2017. Inhibition of bioenergetic metabolism by the combination of metformin and 2-deoxyglucose highly decreased viability of feline mammary carcinoma cells