Small Rna - 16 Regulating Human Ovarian Cancer Cells Biological Characteristics And Platinum Drugs Sensitivity In Vitro Study

Objective To study the role and mechenism of microRNA-16(miR-16) in the proliferation, invasion and apoptosis of ovarian epithelial carcinoma cells in vitro, investigate the expression of microRNA-16(miR-16) and the relationship with cisplatin-sensitive in human ovarian epithelial carcinoma skov-3/DDP cells.Method The skov-3cells and skov-3/DDP cells were seperately transfected with miR-16mimics or negative control RNA(NC) by lipofectamine2000. The expression of miR-16was detected by real-time reverse transcription(RT)-PCR in cells, and western blot was used to detect the expression of vascular endothelial growth factor(VEGF), matrix metalloproteinase-2(MMP-2), B-cell lymphoma2(Bcl-2) protein, and proto-oncogene c-myc protein. Some other methods were used to determine the proliferation and invasion abilities. Tethyl thiazolyl tetrazolium(MTT) was used to determine the cisplatin IC50of the cells. The caspase-3enzyme activity was measured by caspase-3activity assay kit. And the rate of apoptotic cell was detected by flow cytometry method.Results (1)The level of miR-16in cells transfected with miR-16mimics was significantly higher than those transfected with NC (P<0.01);(2)The relative expression of VEGF, MMP-2, Bcl-2and c-myc protein in cells transfected with miR-16mimics were significantly lower than those transfected with NC (P<0.01);(3)miR-16inhibited the proliferation and invasion abilities of skov-3cells dramatically (P<0.05).(4)Twenty-four hours after cultured in serum starvation and hypoxia, the rate of the viable and non-viable apoptotic cells in cells transfected with miR-16mimics was significantly higher(p<0.05). After forty-eight hours, total apoptotic cells in cells transfected with miR-16mimics was significantly more than control cells (P=0.001).(5) The level of miR-16in skov-3/DDP cells was significantly lower than those in skov-3cells, about one-tenth of the latter (P=0.000). And the level of miR-16in the cells transfected with miR-16mimics was660times as high as those transfected with NC (P=0.001);(6)The relative expression of Bcl-2protein and c-myc protein in skov-3/DDP cells were higher than those in skov-3cells, and those in the cells transfected with miR-16mimics were lower than those transfected with NC (P<0.01);(7)After transfected with miR-16mimics, the cisplatin IC50of skov3/DDP cells (14.19±0.06) decreased significantly as compared with those transfected with NC(22.52±0.60)(P=0.002), and the caspase-3enzyme activity unit was much higher (P=0.000).(8) Twenty-four or forty-eight hours after cultured in platinum(20ug/ml), the rate of the apoptotic cells in skov-3group was significantly higher than skov-3/DDP group, and the rate in the cells transfected with miR-16mimics was about1.5fold of the cells transfected with NC (P<0.01)Conclusion (1) miR-16might inhibit the proliferation, invasion of ovarian epithelial cacinoma cells and enhance their sensitivity to apoptotic stimuli through negatively controlling the expression of VEGF、MMP-2and Bcl-2protein;(2) miR-16might enhance the cisplatin-induced apoptosis sensitivity of ovarian epithelial cacinoma cells and play a role in reversal of cisplatin-resistance through negatively controlling the expression of Bcl-2protein, also regulating the expression of c-myc protein cooperately.