Effects of RNA interference silencing RhoC gene upon paclitaxel sensitivity in ovarian cancer cell lines

Abstract

To explore the changes of paclitaxel sensitivity by RNA interference of RhoC gene in ovarian cancer cell lines. The microRNA (miRNA) targeting RhoC gene was designed and synthesized by in vitro transcription and transfected into ovarian cancer cell line SKOV3. The mRNA and protein of RhoC were detected by the means of RT-PCR and Western blot. The changes of paclitaxel sensitivity after interference were examined by methyl thiazolyl tetrazolium (MTf) assay. At the same time, the mRNA of apoptosis gene Caspase-3 and survivin were evaluated by RT-PCR. In SKOV3 cell, the mRNA and protein of RhoC dramatically decreased at 24, 48 and 72 hours post-transfection. The sensitivity of SKOV3 cell line to paclitaxel increased significantly after turning off the RhoC gene. The proapoptotic gene Caspase-3 increased significantly and anti-apoptosis gene survivin dramatically decreased after the combination treatment of paclitaxel and RhoC-miRNA in comparison with paclitaxel alone treatment. The sensitivity of ovarian cancer cell lines SKOV3 to paclitaxel can be enhanced by RNA interfering RhoC gene. The synergistic mechanism is possibly correlated with the up-regulation of Caspase-3 gene and the down-regulation of survivin gene.