To Preliminarily Study On Apoptosis Of Fermented Inonotus Obliquus On Hepatocellular Carcinoma HepG2 Cells

Objective To Preliminarily Study on the apoptosis of fermented Inonotus obliquus(FIO)on hepatocellular carcinoma HepG-2 cells.Methods The contents of polysaccharides,terpenoids and polyphenols in unfermented Inonotus Obliquus(UIO)and FIO were determined by sulfuric acid phenol method and colorimetric method.Hepatocellular carcinoma HepG-2 cells were used as experimental subjects.MTT assay was used to determine the effect of FIO on the proliferation of hepatocellular carcinoma HepG-2 cells.The effect of FIO on morphology of hepatocellular carcinoma HepG-2 cells was observed by HE staining.Flow cytometry was used to determine the effect of FIO on the apoptosis and cell cycle of hepatocellular carcinoma HepG-2 cells.Agarose gel electrophoresis was used to observe the effect of FIO on the DNA of hepatocellular carcinoma HepG-2 cells.Results Compared with UIO,the contents of polysaccharides,polyphenols and terpenoids of FIO were significantly increased(P<0.01).FIO significantly inhibited the proliferation of HepG-2 cells,and its inhibitory effect was positively correlated with the time of administration and the concentration of administration,and compared with the group of UIO,the inhibitory effect of FIO that the concentration was 100 mg/L and the treatment time was 48 h on hepatocellular carcinoma HepG-2 cells was more obvious and in time and dose-dependence(P<0.05).Compared with the control group,the apoptotic morphology of FIO group was observed that the cell spacing of HepG-2 cells was increased,the nuclei were stained,the cells were stained,the apoptotic bodies were formed and the degree of apoptosis increased with the increase in dose and time.Compared with the UIO group,no significant difference was observed by the naked eye.FIO can significantly induce hepatocellular carcinoma HepG-2 cells apoptosis,and with the increase of the concentration,the effect of induced apoptosis enhanced.Compared with UIO,when the concentration of FIO was 200mg/L,400mg(P<0.01).The apoptotic effect on hepatocellular carcinoma HepG-2 cells was significantly higher(P<0.01).Compared with the control group,the cell cycle distribution of HepG-2 cells treated with FIO for 48h was affected that the ratio of GO/G1 phase was obviously decreased(P<0.05)and the ratio of S phase and G2/M phase was markedly decreased(P<0.05).Compared with the control group,the DNA of HepG-2 cells treated with FIO for 48h was cleaved,showing typical trapezoidal apoptotic morphology and compared with UIO,the DNA cleavage of UIO was more obvious.Conclusion FIO can significantly induce apoptosis of hepatocellular carcinoma HepG-2 cells,and compared with UIO,FIO more significantly induces HepG-2 cell apoptosis.