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目的 研究 p5 3基因对胆管癌细胞的作用 .方法 将重组体腺病毒 p5 3转移到人胆管癌细胞 QBC939,用 RT-PCR、克隆形成实验、流式细胞仪、DNA片段化分析等方法对p5 3基因的表达、细胞的生长抑制及机制进行分析 .结果 用Ad- L ac Z进行重组体腺病毒转导效率的检测 ,发现当 MOI为10 0以上时 ,重组体腺病毒可使 90 %以上的培养的人胆管癌QBC939细胞被转导 .用 RT- PCR方法检测 ,在胆管癌QBC939细胞系中 p5 3无表达 .重组体腺病毒能介导外源基因 p5 3在胆管癌 QBC939细胞系中高效表达 .重组体腺病毒介导的 p5 3在 QBC939细胞中表达 ,能抑制 QBC939细胞的生长和集落形成 .流式细胞计数和细胞 DNA L adder,证实p5 3能诱导 QBC939细胞发生凋亡并导致其发生 G1期阻滞 .结论 p5 3基因可能通过诱导肿瘤细胞凋亡及 G1期阻滞在肿瘤的基因治疗方面发挥作用
Objective To study the effect of p5 3 gene on cholangiocarcinoma cells.Methods The recombinant adenovirus p5 3 was transfected into human cholangiocarcinoma cell line QBC939 and the expression of p5 was detected by RT-PCR, clonogenic assay, flow cytometry and DNA fragmentation analysis 3 gene expression, cell growth inhibition and mechanism analysis.Results Recombinant adenovirus transduction efficiency with Ad-L ac Z detection and found that when the MOI is more than 10 0, the recombinant adenovirus can make more than 90% Of cultured human cholangiocarcinoma QBC939 cells were transduced.Expression of p5 3 in the cholangiocarcinoma QBC939 cell line was detected by RT-PCR.The recombinant adenovirus can mediate the expression of the exogenous gene p5 3 in the cholangiocarcinoma QBC939 cell line The recombinant adenovirus-mediated expression of p5 3 in QBC939 cells can inhibit the growth and colony formation of QBC939 cells.Flow cytometry and DNA L adder confirmed that p5 3 can induce apoptosis in QBC939 cells and lead to G1 arrest occurred.Conclusion The p5 3 gene may play a role in the gene therapy of tumor by inducing tumor cell apoptosis and G1 phase arrest