黄卡瓦胡椒素B促进EGFR的降解进而抑制H1975肺癌细胞的增殖和迁移  

Flavokavin B promotes EGFR degradation and inhibits H1975 lung cancer cell proliferation and migration

作　　者：唐颖 李金玲 王科江 杨忠云 李雪森 TANG Ying;LI Jinling;WANG Kejiang;YANG Zhongyun;LI Xuesen(Institute for Cancer Medicine,School of Basic Medical Sciences,Southwest Medical University,Sichuan Luzhou 646000,China)

机构地区：[1]西南医科大学基础医学院肿瘤研究所,四川泸州646000

出　　处：《现代肿瘤医学》2023年第1期17-24,共8页Journal of Modern Oncology

基　　金：国家自然科学青年科学基金项目(编号:81603154)。

摘　　要：目的:研究黄卡瓦胡椒素B对肺癌细胞H1975增殖及迁移的影响并探讨其机制。方法:培养H1975肺癌细胞株,用不同浓度的黄卡瓦胡椒素B处理细胞24、48和72 h后,采用CCK-8法检测黄卡瓦胡椒素B对H1975细胞增殖的影响。Transwell实验检测黄卡瓦胡椒素B对H1975细胞迁移能力的影响。Western blotting检测20μmol/L和40μmol/L黄卡瓦胡椒素B处理16 h后对H1975细胞EGFR-AKT信号通路相关蛋白表达的影响。采用qPCR技术检测黄卡瓦胡椒素B对H1975细胞EGFR mRNA水平的影响。Western blotting检测黄卡瓦胡椒素B对H1975细胞LC3-Ⅰ/Ⅱ蛋白表达的影响。CCK-8实验和Transwell实验分别检测黄卡瓦胡椒素B与EGFR小分子抑制剂联用对H1975细胞增殖和迁移的影响。结果:黄卡瓦胡椒素B在体外能抑制H1975细胞的增殖,且抑制作用呈浓度依赖性。黄卡瓦胡椒素B处理组细胞迁移数明显减少(P<0.05)。Western blotting结果显示,使用黄卡瓦胡椒素B处理16 h后,H1975肺癌细胞的EGFR、p-AKT和AKT蛋白表达水平明显降低(P<0.05),但EGFR mRNA水平无显著改变。当巴佛洛霉素A1(Bafilomycin A1,BAF)或氯喹(Chloroquine,CQ)阻断自噬时,黄卡瓦胡椒素B诱导的对EGFR蛋白的抑制作用被逆转,并且10、20和40μmol/L黄卡瓦胡椒素B可促进H1975细胞LC3-Ⅱ蛋白表达。黄卡瓦胡椒素B与EGFR小分子抑制剂联用能够抑制H1975细胞的增殖和迁移以及p-EGFR和p-AKT的表达。结论:黄卡瓦胡椒素B通过促进EGFR的降解进而抑制H1975肺癌细胞的增殖和迁移。Objective:To study the effect of Flavokavin B on the proliferation and migration of lung cancer cell H1975 and explore its mechanism.Methods:H1975 lung cancer cell line was cultured and treated with different concentrations of Flavokavin B for 24,48 and 72 h.The effect of Flavokavin B on the proliferation of H1975 cell line was detected by CCK-8 method.Transwell assay was used to detect the effect of Flavokavin B on migration of H1975 cells.Western blotting was used to detect the expression of EGFR-AKT signaling pathway related proteins in H1975 cells treated with 20μmol/L and 40μmol/L Flavokavin B for 16 h.qPCR was used to detect the effect of Flavokavin B on EGFR mRNA level in H1975 cells.Western blotting was used to detect the expression of LC3-Ⅰ/Ⅱprotein in H1975 cells.CCK-8 assay and Transwell assay were used to determine the effects of the combination of Flavokavin B and EGFR small molecule inhibitor on the proliferation and migration of H1975 cells.Results:The proliferation of H1975 cells was inhibited in vitro in a concentration-dependent manner.The number of cell migration in treatment group was significantly decreased(P<0.05).Western blotting results showed that the expression levels of EGFR,p-AKT and AKT protein in H1975 lung cancer cells were significantly decreased after treatment for 16 h(P<0.05),but the mRNA level of EGFR in H1975 was not significantly changed.Flavokavin B-induced inhibition of EGFR protein was reversed when autophagy was blocked by Bafilomycin A1(BAF)or Chloroquine(CQ).Moreover,Flavokavin B could promote the expression of LC3-Ⅱprotein in H1975 cells.The combination of Flavokavin B with EGFR small molecule inhibitor can inhibit the proliferation and migration of H1975 cells and the expression of p-EGFR and p-AKT.Conclusion:Flavokavin B can promote EGFR degradation and inhibit the proliferation and migration of H1975 lung cancer cells.

关 键 词：肺癌 黄卡瓦胡椒素B EGFR-AKT信号通路 

分 类 号：R734.2[医药卫生—肿瘤]