Orinidon Inhibits Cell Growth On Human Haptic Carcinoma SMMC-7721 Cells By Induction Of Apoptosis

Objective:To investigate the mechanism of cell growth inhibition and apoptosis inducement of oridonin on human hepatocellular carcinoma SMMC-7721cells.Methods: In vitro,human hepatocellular carcinoma cell line SMMC-7721 was treated by oridonin in different concentrations.The inhibitory effect of cell growth was assayed by MTT assay;cell cycles and apoptosis were analyzed by flow cytometry (FCM);The changes of cell morphology were observed by inverted and confocal microscope,the mRNA expressions of Survivin,Bcl-2,Bax were analyzed by RT-PCR; Western Blot was taken to detect Survivin,Bcl-2,Bax protein expression.Results: MTT assay indicated that the inhibitory effect of orinidon on the proliferation of SMMC-7721 cell was found in dose-and-time dependent manner. Comparing with control group,the cell apoptosis was induced by orinidon at 48 hours,meanwhile,the ratio of G0/G1 phase was increased and the ratio of S phase was decreased, in dose-dependent manner. Marked morphological changes of cell apoptosis were obseved clearly.RT-PCR assay and Western Blot showed that the expression of Bax mRNA and protein were up-regulated and Bcl-2,Survivin mRNA and protein was down-regulated by Orinidon at 48 hours with concentration increased.Conclusion:Oridonin has significant anti-proliferation effects by inducing apoptosis on hepatocellular carcinoma cells in vitro, down regulation of Survivin and Bcl-2 as well as upregualation of Bax expressions may be the important apoptotic inducing mechanisms.