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

Abstract:

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 cells were infected with adenovirus pAdH5-hepaCAM or pAdH5. Methyl thiazolyl tetrazolium（MTT） 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：MTT 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.