Effect Of Curcumin、Curcumol On PI3K/AKT Signal Pathway Of Human Hepatoma Cells SMMC-7721

Objective:To explore the anti-hepatoma and cell apoptosis promotion function mechanism of curcumin and curcumol, which are monomers of Rhizoma Curcumae in ALC from Chinese national expert Zhou Zhongying’s experience prescriptions, so as to illustrate the molecular biological basis for the "cancer toxin" theory proposed by Prof. Zhou, providing evidence for its clinical application and theoretical basis for the research and production of new anti-tumor drugs.Methods:Human hepatoma cells SMMC-7721were cultured in vitro. The effects of curcumin and curcumol on hepatoma cells growth, both from experimental groups were detected by MTT method, cisplatin as positive control group. Hoechst33258fluorescent dye was applied to observe cell apoptosis promotion morphology differences then. Flow cytometer was used to detect cell apoptosis promotion and cell cycle distribution. RT-PCR and Western Blot were applied to find out the effects of curcumin and curcumol on cell PI3K/AKT signal pathway relevant apoptosis protein.Results:MTT showed that curcumin of5-20μg/mL and curcumol of50-200μg/mL had significant effects of inhibiting hepatoma cells proliferation in an evident time and dose dependence manner, with significant difference with the control group. Hoechst33258fluorescent dye indicated that after24h, the cells showed typical apoptosis morphological characteristics:vacuoles in cells, nuclei getting smaller, obviously enhanced fluorescence, appearance of high brightness fluorescent block at nuclei, condensed chromatin, and some cells were in petal, bear’s paw, star or meniscus high brightness block fluorescence shape. FCM showed that after48h, cell apoptosis began and increased dose dependency and compared with the control group, ratio of cells at G0/G1phase increased comparatively and cells of S and G2/M phase reduced, cell cycle was blocked at G0/G1phase. RT-PCR showed that after48h, levels of AKT1and AKT2significantly decreased, expression levels of Caspase9and Caspase3significantly increased dose dependently. Western Blot detection indicated that p-AKT expression decreased, protein activity of PTEN, Caspase9and Caspase3increased, dose dependently after48h. Conclusion:Curcumin and curcumol are two active ingredients in Rhizoma Curcumae from ALC, which can significantly inhibit human hepatoma Cells SMMC-7721proliferation, induce cells apoptosis, of which the mechanism might be related to its inhibition of Go/G1phase, increasing SMMC-7721PTEN expression, inhibiting PI3K/AKT signal pathway activation, decreasing p-AKT protein expression, so as to induce hepatoma cells apoptosis. The study shows that PI3K/AKT signal pathway expression can be one of the mechanisms of their effects.