Isolation, Culture And Identification Of Canine Breast Tumor Cell

Breast cancer is one of the most common tumor diseases in dogs and cats. Caninemammary tumors and human breast tumors have similar pathological features onhistopathology and biology. Without considering life factors, canine mammary tumors andhuman breast tumors occur in very similar ages. Because people love dogs as pets, and livewith together, And dogs and cats share similar living environment with people and havesimilar eating habits, carcinogen exposure is also very similar and so on. Therefore, Caninemammary tumors can serve as a rational animal models of human breast tumors. Tumor cellculture techniques are important basic technology in study carcinogenesis mechanisms,molecular characteristics of the tumor and anti-cancer drug screening. Therefore, thisexperiment followed work on this bas:1. Canine mammary tumor tissue’acquisition, pathological diagnosis and classificationWith reference to the WHO newly released Reference the classification and grading ofbreast cancer in dogs.52cases of canine mammary tumor were collected and were processedclinical histopathological diagnosis by the method of HE. The results show, In52cases ofcanine mammary tumors, There are20cases of benign breast tumors, which occupy38.5%ofthe total cases. There are32cases of malignant breast tumors, which accounting for61.5%ofthe total. This test is the foundation of canine mammary tumor tissue’ digestion, cell’separation and culture, subculture and identification.2. Canine mammary tumor tissue ZNF703quantitative determination testBy real-time PCR technology for8cases of normal canine glandular tissue,20cases ofcanine benign mammary tumor tissue,32cases of canine malignant mammary tumor tissuemRNA in ZNF703(zinc finger protein) expression levels of detection. The results show:Different canine mammary tumor tissue expression of ZNF703gene average varied widely. In8cases, which is normal breast tissue. The expression of samples is from3.07to19.06; In20cases of benign canine mammary tumor tissue, samples’ expression is from10.85to43.16; In32cases of malignant canine mammary tumor tissue samples, the date is from9.43to253.39.Normal breast tissue and benign canine mammary tumors’ ZNF703expression havesignificant difference(p <0.01), the differences is also significant to malignant caninemammary tumor tissue (p <0.01), ZNF703expression of benign canine mammary tumors andmalignant canine mammary tumors also very significant difference (p <0.01). This studyprovide a new reference method for the diagnosis of canine mammary tumors andclassification in molecular biology.3. Isolate, culture and screening tests of canine mammary tumor cellsBy separating, culture and screening the canine mammary tumor cells, we screening andtraining11cases of canine mammary epithelial tumor cell lines which capable passaged more than10times, one in which passages over38generations (CB-23). This study laid thefoundation to follow-up preliminary tests.4. Qualification test to canine mammary tumors CB-23cell lineAccording to the appraisal standard of tumor cells, we have made a series of qualificationtests to CB-23. which were the cell growth curve lines, chromosome analysis, colonyformation, breast cancer-specific antibodies of the CA15-3, and animal tumor inoculation test.The results show: The fast growth of CB-23breast cancer cell lines, and the doubling time isshort; An average of97chromosomes than normal canine chromosome78(39pairs); have ahigh Single cell cloning rate; have strong expression in breast cancer-specific antibodiesCA15-3; Tumorigenic mouse tumor cell inoculation rate of100%; By identifying description,cultured canine mammary tumor cell lines meet the characteristics of canine mammary cancercell, if continue to culture it, it will to be established cell lines. This study laid the basis for thepreliminary study to cancer mechanism and carrying out screening anti-cancer drugs.5. Analysis of the correlation between ZNF703quantitative in malignant breast tumorsand the effect of cell lines cultureBy analysis the correlation between carcinogenic ZNF703gene expression of caninemammary tumor cell lines and generation time，the result showed that ZNF703expression incancer tissues is relatively the higher, passaged cell lines relative to the more, the highestcontent of CB-23cancer cell lines, up to date, has been passed down to38generations, thestudy and analysis of results and may be have benefit to direct screening of cancer cells whichpassaged easy, at the same time, this result may provides a shortcut to choose tumor tissue forcell culture.