基于ROS/PI3K/Akt信号通路探讨异牛肝菌素对人胃癌BGC-823细胞凋亡影响及机制  

Effect and mechanism of iso-suillin on apoptosis of human gastric cancer BGC-823 cells based on ROS/PI3K/Akt signaling pathway

作　　者：徐彦楠[1] 赵俊霞[2] 周娜静[2] 高品 周晨明[3] XU Yan-nan;ZHAO Jun-xia;ZHOU Na-jing;GAO Pin;ZHOU Chen-ming(Department of Teaching and Experiment Center,Hebei Medical University,Shijiazhuang 050017,China;Department of Cell Biology,Hebei Medical University,Shijiazhuang 050017,China;Core Facilities and Centers,Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学教学实验中心,河北石家庄050017 [2]河北医科大学细胞生物教研室,河北石家庄050017 [3]河北医科大学大型科研仪器设备共享服务平台,河北石家庄050017

出　　处：《河北医科大学学报》2022年第10期1121-1126,共6页Journal of Hebei Medical University

基　　金：河北省医学科学研究课题计划(20210149)。

摘　　要：目的基于ROS/PI3K/Akt信号通路探讨异牛肝菌素(iso-suillin)对人胃癌BGC-823细胞凋亡的影响及其机制。方法体外培养人胃癌BGC-823细胞,分别采用7μmol/L、14μmol/L、28μmol/L不同浓度iso-suillin干预48 h,将7μmol/L浓度iso-suillin干预的定义为低剂量组,14μmol/L浓度iso-suillin干预的定义为中剂量组,28μmol/L浓度iso-suillin干预的定义为高剂量组,同时设置对照组(以等量生理盐水干预48 h)。采用Hoechst 33342荧光染色法观察iso-suillin处理后人胃癌BGC-823细胞形态学变化,蛋白免疫印迹法检测iso-suillin处理后人胃癌BGC-823细胞凋亡相关蛋白表达水平,活性氧荧光探针染色法(2,7-dichlorofluorescin diacetate,DCFH-DA)检测iso-suillin处理后人胃癌BGC-823细胞活性氧(reactive oxygen species,ROS)水平,蛋白免疫印迹法检测PI3K/Akt信号通路相关蛋白表达水平。结果Hoechst 33342荧光染色显示随着iso-suillin浓度增加,呈亮蓝色的细胞越来越多,染色质凝聚,出现凋亡小体。与对照组比较,低剂量组、中剂量组、高剂量组凋亡率均升高(P<0.05);低剂量组、中剂量组、高剂量组Caspase-3、Caspase-8、Caspase-9、Bax相对表达量均增加,Bcl-2相对表达量降低(P<0.05)。与对照组比较,低剂量组、中剂量组、高剂量组人胃癌BGC-823细胞ROS水平均升高(P<0.05)。随着iso-suillin浓度的增加,PI3K/Akt信号通路中PI3K、Akt、mTOR蛋白相对表达量均呈降低趋势(P<0.05)。结论异牛肝菌素可诱导人胃癌BGC-823细胞凋亡,其机制可能与上调ROS,下调PI3K/Akt信号通路中PI3K、Akt、mTOR蛋白有关。Objective To explore the effect and mechanism of iso-suillin on the apoptosis of human gastric cancer BGC-823 cells based on the ROS/PI3K/Akt signaling pathway.Methods Human gastric cancer BGC-823 cells were cultured in vitro and treated with iso-suillin at different concentrations of 7μmol/L,14μmol/L,and 28μmol/L for 48 h.The intervention with iso-suillin at a concentration of 7μmol/L was defined as the low-dose group,the intervention with iso-suillin at a concentration of 14μmol/L was defined as the medium-dose group,and the intervention with iso-suillin at a concentration of 28μmol/L was defined as the high-dose group.In the meantime,a control group was set up(intervention with the same amount of normal saline for 48 h).The Hoechst 33342 fluorescent staining method was used to observe the morphological changes of human gastric cancer BGC-823 cells after treatment with iso-suillin,and Western blot was used to detect the expression level of apoptosis-related protein in human gastric cancer BGC-823 cells after treatment with iso-suillin.2,7-dichlorofluorescin diacetate(DCFH-DA)fluorescent probe was used to detect the level of reactive oxygen species(ROS)in human gastric cancer BGC-823 cells treated with iso-suillin,and western blot was used to detect the expression levels of PI3K/Akt signaling pathway-related proteins.Results Hoechst 33342 fluorescent staining showed that an increase in iso-sullin concentration led to more bright blue cells,chromatin condensation,and appearance of apoptotic bodies.Compared with the control group,the apoptosis rate of low-dose group,medium-dose group and high-dose group increased(P<0.05).The relative expression of Caspase-3,Caspase-8,Caspase-9 and Bax increased in low-dose group,medium-dose group and high-dose group,while the relative expression of Bcl-2 decreased(P<0.05).Compared with the control group,the ROS level of BGC-823 cells in low-dose group,medium-dose group and high-dose group increased(P<0.05).The relative expression of PI3K,Akt and mTOR proteins in PI3K/Akt sig

关 键 词：胃肿瘤 异牛肝菌素 BGC-823细胞 

分 类 号：R735.2[医药卫生—肿瘤]