The Effection Of Generation And Apoptosis Of Hepatic Carcinoma Treated By A Retroviral Vector Containing Anti-Sense Htr Gene

Objective : To study the effection of generation and apoptosis of hepatic carcinoma in vivo and vitro treated by a retroviral vector containing anti-sense hTR gene.Methods : we got the recombinant retroviral vector plasmid pLXSN-hTR, which carrying the retro-hTR gene. We except that the transformed hepatic cancer cell mediated by the retrovirus vector could transit anti-sense telomerase RNA, which could combine with the normal telomerase RNA, then stop the expression of telomerase and induce the malignant tumor cell into apoptosis. when the pLXSN-hTR was transferred into packaging cell line PT67 by electroporation, and selected with G418, we got a stable cell line, which could produce retrovirus. After the recombinant retrovirus supernatant was collected the viral titer was determiner with NIH3T3 cell line, we transfected Bel-7402 cell with the newly collected and concentrated retroviral supernatant. The transformed hepatic cancer cell conformed by the retrovirus vector containing pLXSN-hTR could produce anti-sense telomerase RNA. The anti-sense RNA would attach to the normal telomerase RNA and bring down the activity of telomerase, which led to the apoptosis of the hepatic cell line and slew down the growth of the transfected cells of the malignant tumor. We could confirm the existence and expression of the aim gene in Bel-7402 cell through PCR. The therapeutic effects were investigated by cell growth curve. MTT, flow cytometry were used to study the cell apoptosis.A proper model of human liver carcinoma in nude mice was established with Bel-7402 cell. The mice were divided into A, B, C, D, E, F 6 groups at random when the tumor grew to 5mm. The mice were injected intraperitoneally with 20 mg/kg 5-FU, 0.2 ml normal saline solution, 20 mg/kg 5-FU, 20 mg/kg 5-FU, 0.2 ml normal saline solution, 0.2 ml normal saline solution and locally with 0.2 ml BamH1,...