阿霉素诱导肝细胞黏附分子基因下调肾癌细胞细胞分裂周期基因2的表达

Down-regulation of cell division cycle 2 expression in renal carcinoma cell by Adriamycin induced hepatocyte cell adhesion molecule gene

目的:研究阿霉素联合hepaCAM基因对肾癌细胞786-0生物学的影响。方法:阿霉素处理后,用腺病毒向肾癌786-0导入肝细胞黏附分子(Hepatocyte cell adhesion molecule,hepaCAM),以噻唑蓝(Methyl thiazolyl tetrazolium,MTT)测抑制率,流式细胞术(Flow cytometer,FCM)测细胞周期,实时荧光定量聚合酶链反应(Real time quantitative polymerase chain reaction,RT-qPCR)、Western blot检测hepaCAM和细胞分裂周期基因2(Cell division cycle 2,cdc2)的mRNA及蛋白水平。结果:MTT显示:与对照组相比,实验组抑制率显著升高(P<0.01)。FCM证实:实验组出现明显G2/M期阻滞。RT-qPCR及Western blot检测到实验组cdc2 mRNA及蛋白水平显著下调(P<0.01)。结论:阿霉素处理后,导入hepaCAM基因,可引起细胞抑制率显著升高及G2/M期阻滞,这可能与阿霉素诱导hepaCAM下调cdc2的表达有关。

Objective：To investigate the biological effects of Adriamycin combined with hepatocyte cell adhesion molecule （hepaCAM） gene on renal carcinoma cell 786-0. Methods：After treated with Adriamycin, 786-0 ceils were infected with adenovirus pAdH5- hepaCAM or pAdH5. Methyl thiazolyl tetrazolium （MTF） assay was used to observe the inhibition rates of cells in different groups. Flow cytometer（FCM） was used to analyze the changes of cell cycle. Real time quantitative polymerase chain reaction （RT-qPCR） and Western blot were used to detect the mRNA and protein expressions of hepaCAM and cell division cycle 2（cdc2）. Results：MTF assay showed that inhibition rate was markedly increased in ADM/pAdH5-hepaCAM group than in ADM/pAdH5 group and pAdH5- hepaCAM group（P〈0.01）. In FCM assay,remarkable G2/M phase arrest was seen in ADM/pAdH5-hepaCAM group. RT-qPCR and Western blot showed that mRNA and protein levels of cdc2 were down-regulated in ADM/pAdH5-hepaCAM group（P〈0.01）. Conclusions：When gene transfer used after Adriamycin treatment, remarkable growth inhibition and G2/M phase arrest can be observed, which may associate with the down-regulation of cdc2 by Adriamycin induced hepaCAM gene.