The Effect Of Tanshinone â…¡A On Apoptosis And Expression Of MMP-2 And VEGF In Human Ovarian Cancer Cell SKOV3

Objective:To investigate the inhibition function of Tanshinone IIA(Tan IIA) which extract from Tanshinone on human ovarian cancer SKOV3 cell growth, and detect the expression of invasion and metastasis regulatory MMP-2 and VEGF in control group and the experimental group, then explore it's mechanism. For the Tan IIA clinical application and treatment of ovarian cancer provides the theory basis.Methods:SKOV3 cell were treated by Tan IIA with different dose, The proliferation inhibition of Tan IIA to the SKOV3 cell line was examined by MTT assay, cell growth inhibitory rate=(1-ODvalueexperomental group/ODvaluecontrol group)×100%.The experiments were repeated three times, draw cell growth inhibition curve. Half of the inhibitory rate (IC50) was calculated by the software for IC50 calculation.Cell cycles and apoptosis were examined by Flow Cytometry (FCM).4×105cells were inoculated into culture flask of 25cm2,SKOV3 cell was divided into 2 groups randomly,4.0mg/L Tan IIA was added into the experimental groups, according to the length of the time, the experimental groups were divided into 24h,48h.72h group. Collecting the cells of each group, then fixed and stained. Cell cycles and apoptosis was detected by FCM.Immunocytochemical method was used to detect the expression of MMP-2 and VEGF in control group and the experimental group.RT-PCR method was used to detect the expression of MMP-2 mRNA and VEGF mRNA in control group and the experimental group. Results:MTT method showed that:Tan IIA could introduce apoptosis in human ovarian cancer SKOV3 cell, and the growth inhibition was largely dependent of time and concentration. The control group and between groups was statistically significant (P<0.05).FCM showed that after treated by 4.0mg/L Tan IIA for 24,48.72h, the apoptosis of SKOV3 cells markedly increased in G0/G1 phase, and decreased in S phase. Quantitative FCM show the apoptosis rates for 24,48,72h were8.3%,21.9%,28.5%,38.5%. Much more time, much more apoptotic cells. The control group and between groups was statistically significant (P<0.05).Immunocytochemical method showed that after treated by 4.0mg/L Tan IIA for 48h, protein expression of MMP-2 and VEGF declined in experimental group, the fluorescent intensity was abated with the control group, and MMP-2 protein expression decline particularly remarkable. And the difference was statistically significant (P<0.05)RT-PCR showed that after treated by Tan IIA for 48h, the expression level of MMP-2 and VEGF reduced with the increase of drug concentration, significant (p<0.05), and after treated by 4.0mg/L Tan IIA for 24h,48h,72h, the expression level of MMP-2 and VEGF declined, between groups the difference was statistically significant (P<0.05)Conclusion:Tan IIA can significantly inhibit the proliferation of human ovarian cancer SKOV3 cells and induce the cell apoptosis. Declining the expression of MMP-2 and VEGF might be the mechanism that Tan IIA induced invasion in human ovarian SKOV3 cells; The Tan IIA has the potential for treating ovarian cancer that provides theory basis develop anti-tumor drug research.