Effect Of Resveratrol On Expression Of Cytochrome CYP1A1 In Human Hepatoma SMMC-7721 Cells

Objectives Primary liver cancer was one of the most common malignant tumor of digestive tract. It have highly malignant lesions and easily to metastasis. Resveratrol was a kind of polyphenol extraction from plants,such as polygonum cuspidatum and grapes. It has broad pharmacological functions. Resveratrol can protect blood vessels and play an important role in reducing bone calcium level.In recent years, many researchs showed that resveratrol was the effective component for treating lipid metabolism, inflammation and heart disease. The literature reports, Resveratrol has the effect of anti liver cancer, but the mechanism was not clear.The conventional way of antitumor drugs was induction of tumor cell apoptosis and inhibit the proliferation of tumor cells. This experiment researched the influence of Resveratrol on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells in vitro. In order to identify the effective concentration range,CCK-8 was used to detect the proliferation inhibitory effect of Resveratrol on SMMC-7721 at different concentrations. Hoechst 33258 and Annexin V/PI Apoptosis Kit were applied to detect the apoptosis of Resveratrol on SMMC-7721. Real time-PCR technology was used to detection the change of cytochrome CYP1A1 m RNA expressioned in hepatoma SMMC-7721 cells before and after intervention. Western blotting was applied to analyze the expression of cytochrome CYP1A1 protein.Methods :CCK-8 was applied to detect the proliferation inhibitory effect of different concentration Res on SMMC-7721 at different time. The non treated SMMC-7721 cells and the same dose of DMSO respectively to blank group and random control group.Annexin V/PI flow cytometry in effect of 100umol/LRes effect at 72 h on the apoptosis of SMMC-7721 cell rate. Hoechst 33258 was used to observe the effect on the morphology of liver cancer SMMC-7721 cell apoptosis changes in effect of 100umol/L Res effect at 72 h. Real-time PCR analysis of the effect before and after the 100umol/L Res effect on CYP1A1 at 72 h in hepatoma SMMC-7721 cells.Results : CCK-8 results: Res performs a proliferation inhibitory effect on SMMC-7721, the inhibitory rate was 8.38±1.75% after being treated 24 hours with 25μmol/L Res. The inhibitory rate was 15.40±3.32% after being treated 24 hours with 50μmol/L Res. The inhibitory rate was 22.35 ±2.43% after being treated 24 hours with 100μmol/L Res. The inhibitory rate was 15.57±1.73% after being treated 24 hours with 200μmol/L Res. Different concentrations of Res(25μmol/L、50μmol/L、100μmol/L、200μmol/L)function on human hepatoma SMMC-7721 cells after 48 h proliferation inhibition rate of(12.60 + 1.51%, 20.70 + 3.21%, 40.60 + 2.33%, 51.67 + 3.70%)(F=14.03,P=0.03, Different concentrations of Res(25μmol/L、50μmol/L、100μmol/L、200μmol/L)function on human hepatoma SMMC-7721 cells after 72 h proliferation inhibition rate of(15.42 + 3.54%, 24.57 + 3.81%, 60.51 + 3.50%, 65.51 + 2.41%)(F=15.04,P<0.05), The inhibition rate of different concentration Res on the proliferation of hepatocellular carcinoma SMMC-7721 cells was different. In a certain range of concentration, the inhibition effect about Res on the proliferation of hepatocellular carcinoma SMMC-7721 cells depend drug concentration and time.Hoechst-33258 Results: The role of 100umol/L Res after the SMMC-7721 nuclei showed obvious apoptosis morphology. A randomized control group and blank control group have no significant changes in SMMC-7721 nuclei.FCM results: The apoptosis rates of SMMC-7721 after being treated with 100μM/h Res for 72 hours were38.54 %.The apoptosis rates of SMMC-7721 after being treated with blank control for 72 hours are 10.27%.The apoptosis rates of SMMC-7721 after being treated with dose of DMSO for 72 hours are 12.31%.RT-PCR analysis: Expression of CYP1A1 m RNA decreased significantly after 72 h expose in 100umol/L Res. Further testing found that the expression of CYP1A1 protein decreased significantly. Conclusion:In the vitro condition,resveratrol inhibited the proliferation of human hepatoma SMMC-7721 cells and promoted cell apoptosis.The mechanism of resveratrol against liver cancer may be downregulation of CYP1A1 m RNA expression at the transcriptional level and reduction of CYP1A1 protein expression level of translation.