Effect Of SAHA Combined With DDP On Cervical Cancer Cells In Vitro

ObjectiveThrough cultured cervical cancer cell line SiHa in vitro,study on the sensitizing effect of SAHA combined with DDP to SiHa cell,and to explore the mechanism of sensitization. With a view to clinical medication of cervical cancer to provide practical ways.MethodsSiHa cells were treated with 0.25,0.5,1,2,4 and 6μmol/L of SAHA,and 1,2,4,6,8 and 10μg/mL of DDP,separately.Meanwhile the SiHa cells were treated with the vary combination dose of the two drugs. The untreated cells were taken as a control. The effect on proliferation of SiHa cells analyzed with MTT;Cell cycle distribution was detected by flow cytometry;The apoptosis of cells was measured by Annexin V-FITC;P21 and Bax secreted by SiHa cells is determined by RT-PCR and Western bolt. The morphological changes of SiHa cells were observed by inverted phase contrast microscope.ResultsSAHA appears more inhibiting SiHa cells growth effect attending by the increasing dose and time, and there was a synergistic effect when combined with low dose of DDP(P<0.05); SAHA could induce cell apoptosis and arrested cell at G1/G0 phase. When combined with DDP,SAHA induced more severe apoptosis and arrested cell at G1/G0 phase.The combination of two druds increased P21 and Bax protein levels and Mrna levels. Microscopy showed that SiHa cells had a marked proliferation and a high cellular atypia in the control group.But the apoptosis of tumor cells in the treatment group were significantly decreased.A great amount of cell necrosis was observed in the DDP group and the SAHA group.especially in the SAHA+DDP group. ConclusionSAHA can induce apoptosis and cell cycle arrest of SiHa cells, and SAHA has the synergistic effect with DDP. The effect might be associated with up-regulating expression of P21 and Bax mRNA and protein.