Canine mammary Flashcards
1
Q
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
A
2
Q
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?
A
Higher
Skin ulceration, histologic type, nuclear grade, tubular differentiation
Tumors with high levels of TMAS and decrease OS
3
Q
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
A
4
Q
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
A
5
Q
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
A
Shih-Tzu
ER-, PR- and triple negative phenotype and basal like subtype
6
Q
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
A
7
Q
Multiple RT-PCR markers for the detection of circulating tumor cells of metastatic mammary tumors
A
8
Q
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
A
9
Q
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?
A
Prox-1 (markers of human lymphatic vessel) and CD31 (marker of BV)
10
Q
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?
A
transient induction of vimentin (mesenchymal marker)
Invasivness is increased but reversed with prolonged stimulation
11
Q
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
A
12
Q
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
- 5% discrete round cell tumots
- 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
A
13
Q
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
A
COX-2, VEGF-A, VEGF-D
COX-2 with VEGF-A, COX-2 and VEGF-D
14
Q
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 ___
A
decrease cell growth in MCF-7 cells
increase in cell proliferation
15
Q
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
A
16
Q
Pawlowski et al. 2013. Gene expression profiles in canine mammary carcinoma of various grades of malignancy
A
17
Q
A
18
Q
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%
A
64% and 36%
21%
Not different
19
Q
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?
A
Lower
Apoptosis in carcinoma cell lines
Expression is downregulated in canine mammary tumors
20
Q
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?
A
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
21
Q
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
A
22
Q
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
A
inhibited
23
Q
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
A
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
24
Q
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
A
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?
52.6%, 51.7%
Grade 3 (100%)
Grade 1
Tumor cell invasion into lymphatic vesses
Luminal A (44%)
basal-like subtye
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
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
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?
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**
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
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?
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
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**
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**
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
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
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
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?
High expression of GSH
None
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
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
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
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
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
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**
42
Gracanin et al. 2014. Ligand-independent canonical Wnt activity in canine mammary tumor cell lines associated with aberrant LEF1 expression
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.
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
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
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
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
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**
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**
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**
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
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**
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
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**
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**
56
***Borge et al. 2015.* Canine mammary tumors are affected by frequent copy number aberration, including amplification of MYC and loss of PTEN**
## Footnote
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.
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
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
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
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
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.
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
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**
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
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
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
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
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
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
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
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
72
***Raposo et al. 2017. E*xploring 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
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
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
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.
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
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
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
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
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**
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
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
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
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
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.
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%)
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**
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
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
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
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
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
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
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
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.
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
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**
98
Clinical staging in bitches with mammary tumors: Influence of type and histological grade
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
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.
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
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
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
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
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.
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
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
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.
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.
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
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
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
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
114
**Synergistic growth inhibitory effect of deracoxib with doxorubicin against a canine mammary tumor cell line, CMT-U27**
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**
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
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
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**
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.
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.
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
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
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
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%)
124
Canadas et al. 2018. Catechol-o-methyltransferase genotypes are associated with progression and biological behaivour of canine mammary tumors
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?
34 days
109 days
21% and 64%, 28 days and 59 days
100%, 156 days and 180 days
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
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
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
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
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
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
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
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
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
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
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
137
Accuracy of four ultrasonography techniques in predicting histopathological classification of canine mammary carcinomas
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
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
