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目的 :研究 p5 3基因和顺铂联合应用对胆管癌细胞的作用。方法 :将重组体腺病毒p5 3和顺铂联合作用于人胆管癌细胞QBC939,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 -LacZ进行重组腺病毒转导效率的检测 ,发现当MOI为 10 0以上时 ,重组腺病毒可使 90 %以上的培养的人胆管癌QBC939细胞被传导。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。其与顺铂联合应用对QBC939细胞的生长抑制具有明显作用。流式细胞计数证实 p5 3能诱导QBC939细胞发生凋亡并导致其发生G1期阻滞 ,顺铂能诱导QBC939细胞发生凋亡并导致细胞发生明显的G2期阻滞。结论 :p5 3基因能够增加QBC939细胞对顺铂的敏感性
Objective: To study the effect of p5 3 gene combined with cisplatin on cholangiocarcinoma cells. Methods: Combined with recombinant adenovirus p5 3 and cisplatin in human cholangiocarcinoma cell line QBC939, the expression of p5 3 gene, the growth inhibition of cells and its mechanism were analyzed. Results: Ad-Ad-LacZ was used to detect the transduction efficiency of recombinant adenovirus. It was found that more than 90% of cultured human cholangiocarcinoma QBC939 cells were transduced with recombinant adenovirus when the MOI was above 10 0. The expression of p5 3 in QBC939 cholangiocarcinoma cell line was detected by RT-PCR. Recombinant adenovirus can induce high expression of exogenous gene p5 3 in cholangiocarcinoma QBC939 cell line. Recombinant adenovirus-mediated expression of p5 3 in QBC939 cells inhibited the growth and colony formation of QBC939 cells. Its combination with cisplatin has a significant effect on the growth inhibition of QBC939 cells. Flow cytometry confirmed that p5 3 induced apoptosis in QBC939 cells and resulted in G1 arrest. Cisplatin could induce apoptosis in QBC939 cells and lead to significant G2 arrest in cells. Conclusion: p5 3 gene can increase the sensitivity of QBC939 cells to cisplatin