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目的 研究 p1 6基因对胆管癌细胞的作用 .方法 将重组体腺病毒 p1 6转移到人胆管癌细胞 QBC939,对 p1 6基因的表达、细胞的生长抑制及机制进行分析 .结果 用 Ad-Lac Z进行重组体腺病毒转导效率的检测 ,发现当 MOI为 1 0 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 p1 6呈低表达 .重组体腺病毒能介导外源基因 p1 6在胆管癌 QBC939细胞系中高效表达 .重组体腺病毒介导的 p1 6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 Ladder,证实p1 6能诱导 QBC939细胞发生凋亡并导致其发生 G1期阻滞 .结论 p1 6基因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 p1 6 gene on cholangiocarcinoma cells.Methods The recombinant adenovirus p1 6 was transfected into human cholangiocarcinoma cell line QBC939 and the expression of p16 gene and the mechanism of cell growth inhibition were analyzed.Results The expression of p16 gene in cholangiocarcinoma cells was detected by Ad-Lac Z The results showed that when the MOI was above 100, the recombinant adenovirus could transduce more than 90% of the cultured human cholangiocarcinoma QBC939 cells by RT-PCR, The expression of p1 6 in cholangiocarcinoma cell line QBC939 was low.The recombinant adenovirus mediated the high expression of the foreign gene p1 6 in the cholangiocarcinoma cell line QBC939.The recombinant adenovirus mediated p16 expression in QBC939 cells Inhibited the growth and colony formation of QBC939 cells.Flow cytometry and DNA ladder showed that p1 6 induced apoptosis in QBC939 cells and resulted in G1 arrest.Conclusion The p1 6 gene induces tumor cell apoptosis and G1 phase Blocking plays a role in the gene therapy of tumors