苦参碱衍生物ZS10基于PI3K/AKT信号通路诱导人肝癌细胞BEL-7402凋亡  

Synthesis of matrine derivative ZS10 and its mechanism of apoptosis induced by PI3K/AKT signaling pathway in human hepatocellular carcinoma cell line BEL-7402

作　　者：王湘 张森 韩科研 曾攀科 李雪娇 王立升 WANG Xiang;ZHANG Sen;HAN Ke-yan;ZENG Pan-ke;LI Xue-jiao;WANG Li-sheng(Medical College of Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学医学院,广西南宁530004

出　　处：《中国药理学通报》2023年第1期116-124,共9页Chinese Pharmacological Bulletin

基　　金：中央引导地方科技发展专项(No桂科ZY21195012)。

摘　　要：目的探讨新型苦参碱衍生物ZS10抑制BEL-7402细胞增殖,诱导细胞凋亡的信号通路。方法采用有机合成的方法,合成ZS10;采用MTT法,分析在作用时间分别为24、48、72 h时,ZS10对BEL-7402细胞增殖的抑制作用,计算IC 50;采用DAPI染色法,观察BEL-7402细胞状态;采用克隆形成法,观察BEL-7402细胞集落形成情况;采用流式细胞仪观察BEL-7402细胞周期阻滞情况和凋亡情况;采用Western blot法检测PI3K-AKT通路及相关蛋白表达水平。结果获得新型苦参碱衍生物ZS10,并进行了结构表征;MTT结果表明,ZS10作用于BEL-7402细胞48 h,IC 50为(6.62±1.11)μmol·L^(-1);DAPI染色结果表明,给药浓度升高,细胞核发生明显改变;克隆形成结果表明,ZS10明显抑制BEL-7402细胞的集落形成;流式细胞术结果表明,ZS10诱导BEL-7402细胞凋亡并阻滞S期。Western blot结果显示,ZS10在浓度为0~8μmol·L^(-1)时,对PI3K/AKT通路及其相关蛋白具有调节作用,并呈剂量依赖性。与对照组比较,给药组PI3K、AKT、p-AKT和抗凋亡蛋白Bcl-2的表达水平均明显降低,促凋亡蛋白Bax表达水平均明显升高,周期蛋白Cyclin D1和CDK2表达水平均明显降低,迁移蛋白EGFR、N-cadherin、Vimentin表达降低,E-cadherin表达升高。结论ZS10对肝癌细胞BEL-7402的生长、增殖具有抑制作用。Aim To explore the signaling pathway of matrine derivative ZS10 in inhibiting proliferation and inducing apoptosis of BEL-7402 cells.Methods ZS10 was synthesized by organic synthesis.The inhibitory effect of ZS10 on the proliferation of BEL-7402 cells was analyzed by MTT method at the time of 24 h,48 h and 72 h,respectively,and IC 50 was calculated.DAPI staining was used to observe the state of BEL-7402 cells.Clone formation method was used to observe the colony formation of BEL-7402 cells,flow cytometry was used to observe the cell cycle arrest and apoptosis of BEL7402 cells,and Western blot was used to detect the expression level of PI3K/AKT pathway and related proteins.Results MTT results showed that the IC 50 was(6.62±1.11)μmol·L^(-1);DAPI staining showed that the cell state changed significantly with the increase of drug concentration,and the results of colony formation showed that ZS10 significantly inhibited the colony formation of BEL-7402 cells.The results of flow cytometry showed that ZS10 induced S phase arrest and cycle apoptosis of BEL-7402 cells.Western blot showed that ZS10 at the concentration of 0~8μmol·L^(-1) could regulate the PI3K/AKT pathway and its related proteins in a dose-dependent manner.Compared with the control group,the expression of PI3K,AKT,P-AKT and anti-apoptotic protein Bcl-2 significantly decreased,the expression of pro-apoptotic protein Bax significantly increased,the expression of Cyclin D1 and CDK2 significantly decreased,and the expression of EGFR and N-cadherin,Vimentin significantly decreased in the treatment group.The expression of E-cadherin increased.Conclusions Matrine derivative ZS10 can inhibit the growth and proliferation of hepatocellular carcinoma cell line BEL-7402.

关 键 词：苦参碱衍生物ZS10 BEL-7402细胞 迁移 周期 凋亡 PI3K/AKT信号通路 

分 类 号：R284.1[医药卫生—中药学] R329.25[医药卫生—中医学] R345.57R735.7R979.1