Abstract
Canine malignant mammary tumor is a dangerously fatal neoplastic disease with poor survival in female dogs. The aim of this study was to preliminary characterize a novel canine mammary cancer cell line, B-CMT, from canine primary mammary gland tumor, and to utilize it as a cell model for in vitro screening of possible therapeutic drugs. The successfully established cell line, B-CMT, was cultured over 50 passages. B-CMT has a fast proliferation rate, and a population doubling time (PDT) of 33.6 h. The B-CMT cell line lacked human epidermal growth factor receptor-2 (HER-2), estrogen receptors (ER) and progesterone receptors (PR) expression by qRT-PCR. Compared with MDCK cells, CDH1 expression of CMT cell line was significantly decreased or even absent, but GATA3 expression dramatically increased, while TGF-β expression was at a similar level. Interestingly, the B-CMT cell line from canine primary tumor also showed positive hypoxia inducible factor-1α (HIF-1α) results in immunofluorescence (IF), western blot, and qRT-PCR analysis. Ten days post inoculation with EGFP-B-CMT (B-CMT cells stably expressing EGFP), the experimental mice developed palpable soft tissue masses which histologically resembled the canine primary tumor, and was approved to be derived from B-CMT cell line through detection of EGFP by immunohistochemical (IHC) analysis. Moreover, we investigated the cytotoxicity of five drugs to B-CMT cells, and the results showed that rapamycin and imatinib significantly inhibited the proliferation of the cells in vitro within a certain range of concentration. They also induced cell cycle arrest of B-CMT cells at G1 and G2 phase, respectively. In summary, the results of this report showed that B-CMT cell line might serve as a tool for future studies on tumor microenvironment and drug resistance.
Highlights
Canine mammary tumor (CMT) is the most common type of tumor occurring primarily in female dogs, rarely in males, leading to high levels of morbidity in China
The histopathological reports notarized the diagnosis of mammary mixed adenocarcinoma via H&E staining analysis; there was no metastasis in local lymph nodes and distal organs via CT examination
The neoplasm was enveloped with fibrous capsule, and the normal mammary gland structure had been replaced by a large number of proliferative glandular epithelial cells with vigorous division, along with myoepithelial cells
Summary
Canine mammary tumor (CMT) is the most common type of tumor occurring primarily in female dogs, rarely in males, leading to high levels of morbidity in China. Mammary tumors often appear as benign and malignant lesions in dogs, with similar incidence rates. Examination of 1,917 canine mammary biopsies revealed an average annual incidence rate of 16.8%, of which 47.5% were malignant and 47% were benign, respectively. Canine Mammary Cancer Cell Line the average age of dogs having mammary tumors range from 9 to 11 years, with young bitches under 2 rarely having any. Zatloukal et al [2] reported that the average age of dogs with benign and malignant mammary gland tumors was 8.9 and 10 years, respectively. They reported that the percentage of malignant tumors increases correlating with age
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