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目的研究 p16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 p16(Ad-p16)和顺铂联合作用于人胆管癌细胞 QBC939,对 p16基因的表达、细胞的生长抑制及机制进行分析.结果用 Ad-LacZ 进行重组体腺病毒转导效率的检测,发现当MOI 为100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 p16呈低表达.重组体腺病毒能介导外源基因 p16在胆管癌 QBC939细胞系中高效表达,重组体腺病毒介导的 p16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.并显著地抑制该肿瘤细胞的克隆形成能力.流式细胞计数证实 p16能诱导 PBC939细胞发生凋亡并导致其发生 G_1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 G_2期阻滞.结论 p16基因能够增加 QBC939细胞对顺铂的敏感性.
Objective To study the effect of combination of p16 gene and cisplatin on human cholangiocarcinoma cells.Methods The recombinant adenovirus p16 (Ad-p16) and cisplatin combined with human cholangiocarcinoma cell line QBC939 were used to detect the expression of p16 gene and the growth inhibition of cells And mechanism analysis.Results Recombinant adenovirus transduction efficiency was detected by Ad-LacZ and found that when the MOI was above 100, more than 90% of cultured human cholangiocarcinoma QBC939 cells were transduced with the recombinant adenovirus. The expression of p16 was low in cholangiocarcinoma QBC939 cell line by RT-PCR.Expression of p16 was highly expressed in the cholangiocarcinoma QBC939 cell line by recombinant adenovirus.The recombinant adenovirus-mediated p16 was expressed in QBC939 Cells can inhibit the growth and colony formation of QBC939 cells.The combination of cisplatin and cisplatin has a significant effect on the growth inhibition of QBC939 cells and significantly inhibits the clonogenic capacity of the tumor cells.Flow cytometry shows that p16 can induce PBC939 cells apoptosis and lead to the occurrence of G1 phase arrest, cisplatin can induce apoptosis in QBC939 cells and lead to significant cell arrest in G2 phase.Conclusion p16 gene can increase Q BC939 cells to cisplatin sensitivity.