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目的 研究腺病毒介导的多种基因在肝癌细胞中的表达和对肝癌细胞生长的影响。方法 构建含人p5 3、B7 1、GM CSF和IL 2 4种目的基因的重组腺病毒载体 ,感染 3种肝细胞癌细胞系和肝细胞系L0 2 ,运用ELISA、免疫组化、流式细胞仪等方法 ,检测目的基因在肝癌细胞中的表达 ,和对肝癌细胞生长的抑制及诱导其凋亡的作用。结果 肝癌细胞对腺病毒高度易感 ,腺病毒多重感剂量为 5 0 (5 0MOI)时 ,可使 90 %左右的肝细胞癌表达目的基因。除IL 2外 ,其他 3个目的基因都能在肝癌细胞中高效表达。导入多基因后 ,肝癌细胞的生长受到一定程度的抑制 ,并诱导肝癌细胞发生凋亡 ;而L0 2肝细胞的生长却未受影响。结论 腺病毒载体是进行肝癌基因治疗的理想载体 ,由一个腺病毒载体介导的多种目的基因 ,能在肝癌细胞系中高效表达 ,且能抑制肝癌细胞的生长和诱导其调亡。
Objective To study the expression of adenovirus-mediated multiple genes in hepatoma cells and their effects on the growth of hepatocellular carcinoma cells. Methods Recombinant adenovirus vector containing 4 genes of human p5 3, B7 1, GM CSF and IL 2 was constructed and infected with three hepatocellular carcinoma cell lines and L0 2 hepatocellular carcinoma cell line. ELISA, immunohistochemistry, flow cytometry Instrument and other methods to detect the expression of the target gene in liver cancer cells, and the inhibition of liver cancer cell growth and induction of apoptosis. Results The hepatocellular carcinoma cells were highly susceptible to adenovirus. When the adenovirus multi-flu dose was 50 (50 MOI), about 90% of hepatocellular carcinoma cells could express the target gene. In addition to IL 2, the other three genes can be highly expressed in liver cancer cells. After the multi-gene was introduced, the growth of hepatocarcinoma cells was inhibited to a certain degree and induced the apoptosis of hepatoma cells; however, the growth of L0 2 hepatocytes was not affected. Conclusion The adenoviral vector is an ideal carrier for gene therapy of hepatocellular carcinoma. The adenovirus vector can be highly expressed in hepatocellular carcinoma cell lines and can inhibit the growth of hepatoma cells and induce its apoptosis.