Primary Cell Culture And Transplantable Tumor Model Preparation Of Medulloblastoma With SV40Tag

Medulloblastoma, the most common brain tumor in children, is a kind of primitive neuroectodermal tumor (PNETs) of cerebellum and account for approximately 25% of all the tumors in CNS of childhood. It is a major cause of brain cancer mortality. SV40 belongs to polyoma virus. The character of its tumorigenesis is mostly related to T antigen-early protein. Epidemiology shows that it's genomic DNA sequence can be detected in human tumor. But the mechanism of the tumorigenesis of SV40Tag is not clear. The brain tumor model of pTet-on,pTRE2-SV40Tag transgenic mice has been successfully established in our lab, and it was identified to be medulloblastoma.In this research, we successfully operated the primary cell culture of medulloblastoma from transgenic mice model . After handing down from generation to generation, we then cultured them to form monoclonal cell lines. Next, we applied RT-PCR to test the SV40Tag gene on these cell lines and the results were all positive. To further confirm the gene expression , we performed innunohistochemistry. The results showed that the SV40Tag was highly expressed in karyons of these monoclonal cell lines. At the same time, the cells were treated with some anti- tumor medicine, which are commonly used in clinic. And only one drug was tested to have effect on...