基于AMPK/mTOR信号通路研究异常山碱对EC9706细胞能量代谢的调控机制  

Isofebrifuzine regulates energy metabolism of EC9706 cells based on AMPK/mTOR signaling pathway

作　　者：孟丹华 尚艺婉 武颖烁 吴耀松[1] 刘燕[1] 刘俊 李晨旭 陈玉龙[1] MENG Dan-hua;SHANG Yi-wan;WU Ying-shuo;WU Yao-song;LIU Yan;LIU Jun;LI Chen-xu;CHEN Yu-long(Henan Key Laboratory of Signal Transduction,Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学河南省方证信号传导重点实验室,河南郑州450046

出　　处：《中草药》2020年第17期4482-4488,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(81173177);国家自然科学基金面上项目(81373535);河南省高等学校重点科研项目(19A3600-14)。

摘　　要：目的通过观察异常山碱对食管癌EC9706细胞的增殖、凋亡、周期、能量代谢及能量代谢通路相关蛋白表达的影响,探讨其治疗食管癌的分子机制。方法常规培养ECC9706细胞,用MTT法检测细胞活性,筛选药物浓度,选择1、2μg/mL 2个质量浓度;流式细胞术检测异常山碱对EC9706细胞凋亡、周期的影响,能量代谢检测系统检测异常山碱对EC9706细胞能量代谢的影响,Westernblotting检测细胞中哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)、磷酸化乙酰辅酶A羧化酶(p-ACC)、腺苷磷酸激活蛋白激酶(AMPK)蛋白表达。结果不同质量浓度异常山碱作用48h后能有效抑制EC9706细胞增殖,呈剂量依赖性(P<0.01),可以将EC9706细胞阻滞于S期和G2/M期(P<0.05),有效促进细胞凋亡(P<0.05),且明显抑制细胞糖酵解及线粒体代谢能力(P<0.01),与对照组比较,异常山碱组细胞AMPK蛋白表达水平上升,mTOR、p-mTOR、p-ACC蛋白表达水平降低(P<0.05)。结论异常山碱可能通过能量代谢调节EC9706细胞周期、凋亡,抑制EC9706细胞增殖。objective To investigate the molecular mechanism of isofebrifuzine in the treatment of esophageal cancer by observing the effects of isofebrifuzine on proliferation, apoptosis, cycle, energy metabolism and protein expression related to energy metabolism pathway in EC9706 cells. Methods ECC9706 cells were routinely cultured, cell activity was detected by MTT method, drug concentration was screened, and two concentrations of 1 μg/mL and 2 μg/mL were selected, the effect of isofebrifuzine on apoptosis and cycle of esophageal cancer cells EC9706 was detected by flow cytometry. The effect of isofebrifuzine on energy metabolism of EC9706 cells was detected by energy metabolism detection system, and the protein expressions of mTOR, p-mTOR, p-ACC and AMPK in cells were detected by Western blotting. Results The proliferation of EC9706 cells was effectively inhibited in a dose-dependent manner(P < 0.01) after 48 h of treatment with different concentrations of isofebrifuzine, which could arrest EC9706 cells in S phase and G2/M phase(P < 0.05), effectively promote cell apoptosis(P < 0.05), and significantly inhibit cell glycolysis and mitochondrial metabolism(P < 0.01). Compared with the control group, AMPK expression was increased and mTOR, p-mTOR, p-ACC expression was decreased in the treatment group(P < 0.05). Conclusion These results indicated that isofebrifuzine may regulate the cycle and apoptosis of EC9706 cells and inhibit the proliferation of EC9706 cells in esophageal cancer through energy metabolism.

关 键 词：AMPK/mTOR通路 异常山碱 EC9706细胞 能量代谢 增殖 凋亡 周期 

分 类 号：R285.5[医药卫生—中药学]