芪术抗癌方通过STAT3信号通路抑制肝癌细胞Huh-7上皮间质转化的作用机制  

Mechanism of Qizhu Anticancer Prescription in inhibiting epithelial mesenchymal transformation of hepatoma cells Huh-7 through STAT3 signaling pathway

作　　者：孙嘉玲 冯文杏 张仁杰 孙新锋[2] 彭蓝芬 刘兴宁 张卫[2] 蔡本强[2] 周小舟[1,2] SUN Jialing;FENG Wenxing;ZHANG Renjie;SUN Xinfeng;PENG Lanfen;LIU Xingning;ZHANG Wei;CAI Benqiang;ZHOU Xiaozhou(The Fourth Clinical Medical School,Guangzhou University of Chinese Medicine,Shenzhen 518033,China;Shenzhen Traditional Chinese Medicine Hospital,Shenzhen 518033,China)

机构地区：[1]广州中医药大学第四临床医学院,深圳518033 [2]深圳市中医院,深圳518033

出　　处：《中华中医药杂志》2025年第2期612-618,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.82205209);国家自然科学基金面上项目(No.82374531);中国博士后科学基金项目(No.2022M722216);深圳市基础研究专项(自然科学基金)基础研究重点项目(No.JCYJ20210324120405015)。

摘　　要：目的:研究芪术抗癌方对转化生长因子-β1(TGF-β1)诱导Huh-7肝癌细胞上皮间质转化(EMT)的影响,探讨其抑制肝癌细胞EMT的作用机制。方法:以肝癌细胞Huh-7为研究对象,将细胞随机分为空白组,模型组Ⅰ,模型组Ⅱ,芪术抗癌方低、中、高剂量组,索拉非尼组。使用10μg/L TGF-β1诱导Huh-7细胞来构建EMT模型。分别加入相应的含药血清干预细胞后,使用细胞划痕实验、Transwell侵袭实验检测肝癌细胞迁移和侵袭能力的改变,采用细胞免疫荧光和Western Blot检测EMT及信号传导和转录激活因子3(STAT3)信号通路相关蛋白的表达变化。结果:与空白组比较,使用TGF-β1刺激3 d的Huh-7细胞呈梭形改变,细胞间隙增宽,STAT3、p-STAT3、N-cadherin、Vimentin和Snail蛋白表达显著升高(P<0.05,P<0.01),而E-cadherin蛋白表达显著下降(P<0.05)。与空白组比较,模型组Ⅰ和Ⅱ中Huh-7细胞的侵袭和迁移能力均显著增强(P<0.01,P<0.05),STAT3、p-STAT3、N-cadherin、Vimentin和Snail蛋白表达显著升高(P<0.05,P<0.01),而E-cadherin蛋白表达显著下降(P<0.05);与模型组Ⅱ比较,芪术抗癌方低、中、高剂量组和索拉非尼组均可显著抑制TGF-β1诱导的Huh-7细胞的侵袭和迁移能力(P<0.01),芪术抗癌方中、高剂量组均可显著抑制TGF-β1诱导的Huh-7细胞中STAT3、p-STAT3、N-cadherin、Vimentin和Snail蛋白的表达(P<0.05,P<0.01),升高E-cadherin蛋白表达(P<0.01)。细胞免疫荧光结果显示,与空白组比较,模型组Ⅰ和Ⅱ细胞质中STAT3、p-STAT3表达增多,促进p-STAT3入核;与模型组Ⅱ比较,芪术抗癌方低、中、高剂量组和索拉非尼组均可抑制细胞质中STAT3、p-STAT3表达和p-STAT3入核。结论:芪术抗癌方可能通过调控STAT3信号通路来抑制TGF-β1诱导肝癌细胞Huh-7的EMT进程,从而发挥抗肝癌的作用。Objective:To study the effect of Qizhu Anticancer Prescription on transforming growth factor-β1(TGF-β1)induced epithelial mesenchymal transformation(EMT)of Huh-7 hepatoma cells,and to explore the mechanism of inhibiting EMT in hepatoma cells.Methods:HCC cells Huh-7 were randomly divided into blank group,model group I and model group II,Qizhu Anticancer Prescription low,medium,high dose groups,Sorafenib group.The EMT model was constructed using 10μg/L TGF-β1 induced Huh-7 cells.After the corresponding drug-containing serum was added to Huh-7 cells,cell wound healing and Transwell invasion test were used to detect the changes of migration and invasion ability of hepatoma cells.Cellular immunofuorescence and Western Blot were used to detect the expression changes of proteins associated with EMT and signal transducer and activator of transcription 3(STAT3)signaling pathway.Results:Compared with the blank group,after 3 days of TGF-β1 stimulation,Huh-7 cells showed fusiform changes and the cell gap widened,the expressions of STAT3,phosphorylated p-STAT3,N-cadherin,Vimentin and Snail proteins were significantly increased(P<0.05,P<0.01).The expression of E-cadherin protein decreased significantly(P<0.05).Compared with blank group,the invasion and metastasis ability of Huh-7 cells in model group I and I were significantly enhanced(P<0.01,P<0.05),the protein expressions of STAT3,p-STAT3,N-cadherin,Vimentin and Snail in model groups I and II were significantly increased(P<0.05,P<0.01),while the protein expressions of E-cadherin were significantly decreased(P<0.05).Compared with model group II,Qizhu Anticancer Prescription low,medium and high dose groups and Sorafenib group could significantly inhibit the invasion and metastasis ability of TGF-β1 induced Huh-7 cells(P<0.01).Qizhu Anticancer Prescription medium and high dose groups significantly inhibited the expression of STAT3,p-STAT3,N-cadherin,Vimentin and Snail proteins in TGF-β1 induced Huh-7 cells(P<0.05,P<0.01),and increased the expression of E-cadherin protein

关 键 词：芪术抗癌方 肝癌细胞Huh-7 上皮间质转化 信号传导和转录激活因子3信号通路 转化生长因子-Β1 

分 类 号：R285[医药卫生—中药学]