| Objective:To investigate the cell regulation of Ginsenoside Compound K on SMMC-7721 and the effect to apoptosis of SMMC-7721 through the mechanism of TGF-β1/Smads signaling pathway provides theoretical basis for the study on the treatment of liver cancer by Ginsenoside CK.Methods:Cultivate human liver cancer SMMC-7721 cells which were divided into control group,Ginsenoside CK group(20 μmol/L,40 μmol/L,60 μmol/L),MTT was used to detect the growth rate of Ginsenoside CK in human liver cancer SMMC-7721.The apoptosis effect of Ginsenoside CK on human liver cancer SMMC-7721 was detected by Hoechst staining and Annexin V FITC/PI.Immunocytochemistry staining method is to detect the expression of Smad4 and Tβ RⅡ which are pathways related proteins.What’s more,the protein expression of TGF-β1,Tβ RⅡ,Smad2/3,Smad4,Smad7 and Caspase-3 was detected by Western Bot.Results:1.The results showed that Ginsenoside CK inhibited the proliferation of human SMMC-7721 cells in a dose-and-time dependent manner,with growth rates(%)of 88.0±8.1,57.0±6.6,34.2±5.6,10.0±5.9 at 48 h;2.The result of HOECHST staining showed that Ginsenoside CK induced the apoptosis of SMMC-7721 and Cleaved-Caspase3 is promoted which is detected by Western Blot;3.When SMMC-7721 is cultivated by Ginesnoside CK for 48 h,the result showed that the the apoptosis rates(%)are7.2±1.2、23.5±2.1、46.2±1.2 by Annexin V FITC/PI;4.The results of ICC showed that Smad4 protein expression is inhibited with the concentration of Ginsenoside CK growing;5.By western blot,the expression of TGF-β1,TβR Ⅱ,Smad2/3 and Smad4 protein is inhibited with the concentration of Ginsenoside growing while the expression of Smad7 is elevated.When human recombinant protein TGF-β1(10 ng/ml)is added,TGF-β1+CK group has a higher expression of Smad2/3 and Smad4 and a lower Smad7 expression.Conclusion:Taken together,our results show that Ginsenoside CK induces apoptosis of SMMC-7721 and the main mechanism is related to TGF-β1/Smads signaling pathway. |
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