Raf Kinase Inhibitor Enhanced Chemotherapy Sensitivity In Cervical Cancer Cells

Objective:To investigate the effects of up-regulation of Raf kinase inhibitor protein (RK1P) on the chemosensitivity of cervical cancer Hela cells.Methods:Eukaryotic expression plasmid pcDNA3.1(+)-ssRKIP containing human overall length RKIPcDNA was transfected into cervical cancer Hela cell by lipofectin assay, establishing a stable cell line containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of different concentrations and intervals of time, the effect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP can inhibit apoptosis in Hela cells induced by cisplatin.Results:The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After different concentrations of cisplatin treatment cells for24,48,72h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells(P<0.05). With5μg/ml cisplatin treatment for24h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis (23.2±0.24)%than non-transfected cells (12.4±0.31)%and empty vector-transfected cells (13.4±0.47)%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was (5.7±0.12)%. which was still higher than those of the non-transfected cells(2.9±0.21)%and empty vector-transfected cells(3.6±0.08)%.Conclusions:Higher expression of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.