Mechanisms Of The Apoptosis Induced By Sinomenine In Human Liver Cancer Cell Line HepG2

Objective: Treatment on hepatoma cells hepG2 by using different concentrations of sinomenine, this paper studies the inhibitory effects of sinomenine on HepG2 from morpHological structure and inhibition rate of cells which provide clinical application with theoretical basis.Then by testing the variation of survivin effected by different sinomenine concentration, this paper shows the relationship between the expression of survivin induced by sinomenine and sinomenine mediated HepG2 in G2/M period, to discuss the function of survivin in inducing apoptosis of sinomenine.Methods: To observe and take pHoto by inverted microscope, SRB Test, AO/EB, Hoechst fluorescence staining and to test the toxicity of SN on hepatoma cells hepG2 and function of inducing apoptosis by taking pHotos using fluorescence microscope and other technical test. Flow cytometry is used to test cell cycle distribution and apoptosis rate, Quantitative RT-PCR is applied to test the expression of survivin mRNA, Western blot is used to test the expression of survivin which is related to apoptosisResult: SN could significantly inhibit the growth of Hep G2, and it has time -and concentration-dependent manner. SN Half growth concentration (IC₅₀) in 24h, 48h, 72h are1.44,1.21,0.39 mM. After 24hours function of SN at concentration 0.2 mM～6.4mM, Hoechst nuclear stain, HepG2 nuclear is fracture, fragmented and weekly changed, showed a characteristic of typical apoptosis. FCM shows that cells was blocked at G2/M period, it was increased with medicine concentration. The result of Quantitative RT-PCR shows that sinomenine inhibits the expression of HepG2 survivin mRNA. Western blot test shows that sinomenine can inhibit the expression of HepG2 survivin, as the time and concentration are increased, the expression of survivin protein will decreased, it also highly expressed after 24 hours function of SN.Conclusion:(1) SN has a strong function that can inhibit the proliferation of cells in liver cancer. It has time and concentration dependent manner.(2) SN has a function of inducing apoptosis of liver cancer cell line HepG2, it has It has time -and concentration- dependent manner.(3) Sinomenine prohibited proliferation during cell cycle G₂/M(4) Survivin plays an vital role in inducing apoptosis of sinomenine, there is a negative correlation between expression and concentration.(5)The expression of survivin induced by sinomenine may be a phenomenon in early stage which has nothing to do with sinomenine mediated HepG2...