The Studies On Suppression Of Growth,Induction Of Apoptosis And Its Mechanisms Of Human Ovarian Cancer Cell By Oridonin

Objective To investigate the effect of oridonin which extracts from Donglingcao on the suppression of growth and induction of apoptosis in human ovarian cancer SKOV3cell,and explore its mechanism.Methods The effect of oridonin separate, and oridonin combined cisplatin on ovarian cancer cell SKOV3by MTT assay; The changes of morphology and microstructure were observed by inverted microscope and electron microscope; Cell cycles and apoptosis rate were examined by Flow Cytometry (FCM); the change of methylation status of CpG island in hMLHl gene promoter region was analyzed by MSP and the change of expression of bcl-2,bax,survivin and hMLH1mRNA was detected by QRT-PCR in SKOV3cell.Results①MTT assay showed that compared with the control group, oridonin could significantly inhibit the proliferation of SKOV3cells in time-dose-dependant manner (P<0.05),and the IC50of SKOV3cells treated with oridonin for24,48,72hours were45.22,33.85,22.48μmol/L respectively; the inhibitive effect of oridonin combined cisplatin was more obvious than either one used separate.②Inverted microscope observeed that cell growth was obviously inhibited.③Apoptosis morphological changes about vacuolar degeneration, chromatic agglutination and nuclear condensation was detected by electron microscope in oridonin-treated SKOV3cells.④Cell cycle analysis indicated that the oridonin-treated cells were arrested at the S and G2/M phases.⑤The results of FCM double-stained represented the apoptosis rate of SKOV3cells treated with10,20and30μmol/L oridonin for48hours were(9.13±0.91)%,(18.89±1.79)%,(36.11±3.01)%respectively,and the apoptotic rate increased significantly compared with control group,the difference was statistically significant(P<0.05);⑥MSP result that the CpG island in hMLHl gene promoter region in SKOV3cells showed hemimethylation, after being treated with oridonin, methylation specificity strip of CpG island in hM1H1gene promoter bate unconspicuously and unmethylation specificity strip enhanced;⑦QRT-PCR showed that Oridonin obviously decrease the expression of bcl-2and survivin but increase the expression of bax and hMLHl mRNA,and the difference was statistically significant compared with control group (P<0.05). Conclusion①Oridonin can significantly inhibit the proliferation and induce the cell apoptosis in human ovarian cancer SKOV3cell;②The expression of bcl-2and survivin downregulated and bax upregulated might be the one of the mechanism that Oridonin induced apoptosis in human ovarian cancer SKOV3cell; the another of the mechanism that Oridonin induced apoptosis of human ovarian cancer SKOV3cells may through demethylation effect on hMLH1gene.