摘要
目的:探讨栓菌酸对肝癌HepG2细胞株耐索拉非尼(Sorafenib)的作用及其机制。方法:通过浓度梯度递增法建立对索拉非尼获得性耐药细胞株HepG2/Sora;Western Blot法检测HepG2、HepG2/Sora细胞内AKT、p-AKT及c-RAF蛋白的表达;MTT法检测栓菌酸细胞毒性,同时检测栓菌酸和索拉非尼联用后对HepG2/Sora的活性影响,Western Blot法检测低细胞毒性浓度5、10μmol/L栓菌酸联合索拉非尼对HepG2/Sora细胞内AKT、p-AKT、mTOR、p-mTOR、c-RAF、VEGFR2的蛋白表达。结果:建立了索拉非尼耐药细胞株HepG2/Sora,耐药指数为4.7;HepG2/Sora细胞中p-AKT及c-RAF较HepG2细胞显著上调(P<0.01);低细胞毒性浓度5、10μmol/L栓菌酸联合索拉非尼作用HepG2/Sora细胞后,药物敏感性增强,联合用药指数均小于1,具有协同作用,逆转倍数分别为5.17、7.55;Western Blot结果显示,栓菌酸联合索拉非尼可以显著下调PI3K/AKT/mTOR信号通路相关蛋白AKT、p-AKT、mTOR、p-mTOR及索拉非尼作用靶点c-RAF、VEGFR2的蛋白表达(P<0.01)。结论:栓菌酸能有效逆转HepG2/Sora耐药细胞对索拉非尼的耐药,其机制可能与抑制PI3K/AKT/mTOR信号通路有关。
Objective:To investigate the effect of trametenolic acid B(TAB)on sorafenib resistance in l hepatoma cell line HepG2 and its mechanism.Methods:The acquired sorafenib resistant cell line HepG2/Sora was established by concentration gradient increasing method.Western blotting was used to detect the protein expressions of protein kinase B(AKT),phosphor-AKT(p-AKT),and C-rapidly accelerated fibrosarcoma(C-RAF)in HepG2 and HepG2/Sora cells.Methylthiazolyl tetrazolium(MTT)assay was used to detect the cytotoxicity of TAB and the effect of TAB combined with sorafenib on HepG2/Sora activity.Moreover,Western blotting was also used to detect the protein expressions of AKT,p-AKT,mammalian target of rapamycin(mTOR),phosphor-mTOR(p-mTOR),C-RAF,and vascular endothelial growth factor receptor 2(VEGFR2)in HepG2/Sora cells at low cytotoxic concentration(5 and 10μmol/L)of TAB combined with sorafenib.Results:The sorafenib resistant cell line HepG2/Sora was established,with a resistance index of 4.7.As compared with HepG2 cells,the p-AKT and C-RAF in HepG2/Sora cells were significantly up-regulated(P<0.01).The drug sensitivity of HepG2/Sora cells was increased after the treatment of TAB combined with sorafenib at low cytotoxic concentration(5 and 10μmol/L),and the combined drug index was less than 1,showing the synergistic effect.The reversal indexes were 5.17 and 7.55,respectively.Western blot results showed that TAB combined with sorafenib significantly down-regulated the expressions of PI3 K/AKT/mTOR signaling pathway-associated proteins including AKT,p-AKT,mTOR,and p-mTOR,and the targets of sorafenib such as C-RAF and VEGFR2(P<0.01).Conclusion:TAB effectively reverses the sorafenib resistance in HepG2/Sora cells,and its mechanism may be related to the inhibition of PI3 K/AKT/mTOR signal pathway.
作者
袁园
汪鋆植
邓李蓉
高元喜
叶汪阳
张晓兰
邓改改
Yuan Yuan;Wang Junzhi;Deng Lirong;Gao Yuanxi;Ye Wangyang;Zhang Xiaolan;Deng Gaigai(Hubei Provincial Key Laboratory of Natural Products Research and Development,China Three Gorges University,Yichang 443002;Yichang Hospital of Traditional Chinese Medicine,Yichang 443002;School of Basic Medical Sciences,Guangxi Medical University,Nanning 530021)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第3期76-81,共6页
Pharmacology and Clinics of Chinese Materia Medica
基金
湖北省卫生计生委中医药科研项目(编号:ZY2021M087)
关键词
栓菌酸
索拉非尼
肝癌
耐药
trametenolic acid B
sorafenib
liver cancer
drug resistance
作者简介
通信作者:汪鋆植,博士,教授,主要从事土家族药物活性评价和质量控制研究,E-mail:horsedog@163.com;袁园,硕士,主要从事药理学研究,E-mail:470803906@qq.com。
