Effects Of Heshuxiaoji Decoction On The Proliferation And Apoptosis Of HepG2 Cell Line By Regulating NS5ATP13 Positive Feedback

Objective: Heshuxiaoji decoction is a recipe that Professor Wang Jing of the Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University has experienced in the treatment of primary liver cancer for many years.In long-term clinical practice,we found that this prescription can significantly slow down the progression of liver cancer,improve the survival rate of patients,and has a clear clinical effect.However,the mechanism of its treatment of HCC remains unclear.In this study,the human hepatoma cell line HepG2 was used as the research object.In vitro experiments were conducted to explore the effect of Heshuxiaoji decoction on the proliferation and apoptosis of HepG2 cells.To reveal the potential mechanism of Heshuxiaoji decoction in the treatment of liver cancer,provide a scientific theoretical basis for the application of Heshuxiaoji decoction,and provide new ideas for the treatment of liver cancer with integrated traditional Chinese and Western medicine.Methods: 1.HepG2 cells were cultured in vitro,and the effect of different concentrations of Heshuxiaoji decoction on the activity of liver cancer cells was detected by CCK-8 assay,and the appropriate drug concentration was screened for subsequent experiments.2.Combined CCK-8 experiment and plate cloning experiment to detect the effect of each group and Heshuxiaoji decoction on the proliferation of HepG2 cells.3.Combined application of cell scratch test,Transwell invasion test and migration test to detect the effect of Heshuxiaoji decoction on the invasion ability and migration ability of HepG2 cells.4.Using Annexin v-FITC/PI flow cytometry to detect the effect of Heshuxiaoji decoction on HepG2 cell necrosis and apoptosis.5.Western blot was used to detect the effect of Heshuxiaoji decoction on the expression levels of NS5ATP13,Bcl 2,Bax and AKT/GSK/MTOR-related pathway proteins.Results: 1.Compared with the control group,Heshuxiaoji decoction can significantly reduce the activity,proliferation rate,cell healing rate,invasion rate and migration rate of HepG2 cells(P < 0.05),and its inhibitory ability is concentration-dependent.2.The results of Annexin v-FITC/PI flow cytometry showed that Heshuxiaoji decoction could promote the apoptosis of HepG2 cells in a concentration-dependent manner(P < 0.05).3.The results of Western blot showed that after the intervention of Heshuxiaoji decoction,the expression of Bax protein in HepG2 cells could be up-regulated,and the protein expression levels of Bcl 2 and p-Akt,p-GSK and p-MTOR could be down-regulated(P <0.05).4.Real-time PCR results showed that Heshuxiaoji decoction could reduce the expression of NS5ATP13 at m RNA and protein levels in HepG2 cells(P <0.05).Conclusion: Heshuxiaoji decoction can inhibit the proliferation and induce apoptosis of HepG2 cells,and the mechanism may be related to Heshuxiaoji decoction inhibiting the expression of NS5ATP13 and then inhibiting the activation of AKT/GSK/mTOR signaling.