摘要
目的:探讨和枢消积方通过调节三氧化二砷反式激活蛋白3(AsTP3)对人肝癌细胞SMMC-7721增殖、凋亡的作用机制。方法:体外培养SMMC-7721肝癌细胞,采用CCK8法筛选出和枢消积方最适浓度,联合CCK8法与平板克隆形成实验检测细胞增殖,细胞划痕实验及Transwell小室实验检测细胞迁移、侵袭能力,流式细胞实验检测细胞凋亡,qRT-PCR检测细胞AsTP3 mRNA水平,Western Blot检测细胞中AsTP3、AKT、GSK-3β、mTOR蛋白相对表达量及磷酸化水平。结果:与对照组比较,和枢消积方能抑制SMMC-7721细胞增殖、迁移、侵袭及克隆能力,促进SMMC-7721细胞凋亡(P<0.05);和枢消积方可下调SMMC-7721细胞内AsTP3、P-AKT、P-GSK-3β、P-mTOR、Bcl2的mRNA表达及蛋白水平并上调Bax蛋白的表达。结论:和枢消积方能抑制SMMC-7721细胞增殖,促进其凋亡,其机制可能与下调AsTP3表达从而抑制AKT/GSK-3β/mTOR信号通路有关。
Objective:To investigate the effect of Heshuxiaoji formula on the proliferation and apoptosis of human hepatoma cell SMMC-7221 by regulating the arsenic trioxide-activated protein 3(AsTP3).Methods:SMMC-7721 hepatoma cells were cultured in vitro and the optimal concentration of Heshuxiaoji formula was selected by using CCK8.CCK8 and plate colony-forming assay were used to detect cell proliferation activity,cell migration by cell wound scratch assay,cell invasion by Transwell,and cell apoptosis by Flow cytometry.qRT-PCR and Western Blot were used to detect the expression of apoptosis-related genes such as AsTP3.Results:Compared with the control group,Heshuxiaoji formula could inhibit the proliferation,migration,invasion,and cloning of SMMC-7721 cells,and promote the apoptosis of SMMC-7721 cells.The Heshuxiaoji formulation may attenuate the messenger RNA and protein expression levels of AsTP3,p-AKT,P-GSK-3β,P-mTOR,and Bcl2,while concurrently augmenting the protein expression of Bax within SMMC-7721 cells.Conclusion:Heshuxiaoji decoction can inhibit the proliferation of SMMC-7721 cell and promote their apoptosis,which may be related to the down-regulation of AsTP3 expression and the inhibition of AKT/GSK-3β/mTOR signaling pathway.
作者
李玲
汪静
朱晓宁
张玉蓉
尹玥
LI Ling;WANG Jing;ZHU Xiao-ning(Department of Hepatobiliary Diseases,The Affiliated Traditional Medicine Hospital of Southwest Medical University,Luzhou Sichuan,646000,China)
出处
《中西医结合肝病杂志》
CAS
2024年第2期131-135,139,共6页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
四川省科学技术厅重大科技专项(No.2019YFS0018)
泸州市人民政府-西南医科大学科技战略合作项目(No.2019LZXNYDZ12)
西南医科大学附属中医医院国家中医临床研究基地建设单位科研项目(西南医大中医院[2020]33号)
西南医科大学-西南医科大学附属中医医院联合项目(No.2020XYLH-037,2018XYLH-030)。
作者简介
通讯作者:汪静,E-mail:lywj68@126.com。
