Apoptosis Mechanism Of Human Gastric Cancer Cell Line SGC-7901 Induced By AHVAC-Ⅰ

Objective: The study is designed to determine the roles of Agkis-trodon halys venom antitumor component I(AHVAC-Ⅰ)in human gastric cancer cell line SGC-7901 proliferation and oxidative stress and underlying mechanism.Methods: To determine the roles of AHVAC-Ⅰ in human gastric cancer cell,the cells were divided into negative control group and different concentration experimental groups treated with AHVAC-Ⅰ(5,10,20,30,40 μg/m L).Cell proliferation was evaluated by CCK8 assayt.In addition,HE staining was used to observe the morphological changes of human gastric cancer cell.According to the abrove results of CCK8,the optimal AHVAC-1(5,10,20 μg/m L)concentrations were selected for subsequent experiments.Flow cytometry and immunocytochemistry staining were used to analyze the effects of AHVAC-Ⅰ on apoptosis.Furthermore,we used the assay kits to assess intracellular ROS,Mn-SOD levels and the total antioxidant capacity to determine the oxdaitve stress in SGC-7901,which were treated with AHVAC-Ⅰ.Western blot and Real-time PCR were used to detect the expression levels of Bax and Bcl-2 to determine the molecular pathway of AHVAC-Ⅰ-induced apoptosis in SGC-7901 cells.Results: 1、After treatment with different concentrations of AHVAC-Ⅰ(5,10,20,30,40 μg/m L)for 24 h,the results of CCK-8 showed that AHVAC-Ⅰ inhibited the growth and proliferation of SGC-7901 cells in a concentration-dependent manner.The half-inhibitory concentration IC50 is 21.878 μg/m L according to the modified method.While,the results of HE staining showed demonstrated morphological changes in SGC-7901,which treated with 20μg/m L AHVAC-Ⅰ.Then,the optimal AHVAC-1 concentraion was 20 μg/m L.2、The flow cytometry and immunohistochemistry is used to detect the cells at different concentrations and times.The results showed that the apoptosis rate of SGVAC-I cells treated with AHVA-I for 24 h increased with the concentration of AHVAC-Ⅰ at 5,10,20 μg/m L,showing a dose-dependent effect.3、After SGC-7901 cells were treated with different concentrations of AHVAC-Ⅰ(5,10,20μg/m L)for 24 h,the activities of active oxygen and Mn-SOD increased,while the total antioxidant capacity of cells decreased gradually and showing a dose dependency effect.4、 The results of Western blot and Real-time PCR showed that the expression level of anti-apoptotic protein Bcl-2 was significantly decreased,while the expression of proapoptotic protein Bax was significantly increased.Conclusion: 1、AHVAC-Ⅰ induced apoptosis of human gastric cancer cell line SGC-7901,and the apoptosis rate is dose-dependent manner;2、AHVAC-Ⅰ increased the oxidative stress in human gastric cancer cell line SGC-7901 by inhibiting the expression of anti-apoptotic protein Bcl-2.