Verbascoside Affect The Proliferation And Autophagy Of HCC Cells By Regulating The JNK Signaling Pathway

Objective Study the effect of verbascoside on proliferation and autophagy of hepatocellular carcinoma cells in vitro and explore specific molecular mechanism,and furthermore to explore the correlation between the mechanism and the JNK signaling pathway,so as to provide experimental and theoretical basis for the further development of anti-HCC drugs with verbascoside as the main active ingredients component.Methods a.HepG2 and 7404 liver cancer cells were selected as the cell lines for this experiment,and the effects of different concentrations of verbascoside on the morphological changes of hepatocellular carcinoma cells were observed under an inverted phase contrast microscope.b.CCK8 method was used to detect the effects of different concentrations of verbascoside on the proliferation of hepatocellular carcinoma cells after different periods of action,and the inhibition rate was calculated to determine the optimal concentration and time of action of verbascoside in inhibiting the proliferation of hepatocellular carcinoma cells.c.HCC hepatocellular carcinoma cells were set as blank control group,the optimal concentration of verbascoside group and inhibitor group(JNK inhibitor SP600125),and the expressions of JNK p-JNK and autophagy proteins(Beclin1 and LC3)in hepatocellular carcinoma cells in each group were detected by Western Blot method.Results a.With the increase of the concentration and action time of verbascoside,the morphology of HCC cells was observed under the microscope to gradually change,including adherence and loose arrangement,rounded cell morphology and even death,indicating that verbascoside could affect the growth of hepatocellular carcinoma cells.b.With the control group compared,the optimal concentration of verbascoside in inhibiting the proliferation of liver cancer cells was 50?M and the time was 24 h,showing a significant difference(P < 0.05).c.The expression of JNK and p-JNK protein in HepG2 hepatocellular carcinoma cells of the optimal concentration and the inhibitor group was significantly down-regulated,and the expression of p-JNK protein in 7404 hepatocellular carcinoma cells was significantly down-regulated,compared with the blank control group,after the treatment of hepatocellular carcinoma cells of each group with the optimal concentration of chaetoside glycosides(p < 0.05).The expression levels of Beclin1 and LC3 autophagy proteins in hepatocellular carcinoma cells of the two groups were significantly up-regulated(P < 0.05),while the expression levels of Beclin1 and LC3 autophagy proteins in the two groups with the optimal concentration of triosyllabine glycosides and inhibitors were not significantly changed(P>0.05).Conclusion a.With the increase of the concentration of verbascoside and the longer the effect time,the more significant the effect was on the survival and proliferation of liver cancer cells.2.Verbascoside can inhibit the proliferation of hepatocellular carcinoma cells and promote autophagy in a concentration and time dependent way.c.By interfering with the activation of the JNK signaling pathway,the glycosides of chaetosides can regulate the expression of proliferation and autophagy proteins,and thus inhibit the proliferation of hepatocellular carcinoma cells.