腺病毒介导的p53对胆管癌细胞生长的抑制作用

Inhibitory effects of recombinant adenoviruses p53 transduction on human cholangiocarcinoma cell line

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

AIM To evaluate the inhibitory effect of p53 on human cholangiocarcinoma cell line. METHODS The effects of recombinant adenoviruses p53 (Ad p53) on the growth of QBC939 cells in vitro were analyzed. RESULTS The QBC939 cell line transducted with recombinant adenoviruses LacZ ( Ad LacZ) at MOI of 100 or greater exhibited more than 90% transduction rate. p53 expression was negative in QBC939 cell but high levels of p53 expressions were observed in QBC939 cell line infected with Ad p53. The growth rates of QBC939 cells by Ad p53 at MOI of 100 inhibited reduced by 19.3%(6 day) or 24.4%(8 day) respectively (MOI of 100), compared with those of the Ad LacZ infected cells. Colony formation in these Ad p53 infected cells greatly decreased versus Ad LacZ infected cells. The Ad p53 transfected cells induced QBC939 cell apoptosis and G1 blockade, which was confirmed by the flow cytometry and DNA Ladder. CONCLUSION The suppression effects mediated by expression of the exogenous p53 on tumor cells result mainly from apoptosis and G1 blockade.