| 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 |
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| 引用本文： | 唐颖,李金玲,王科江,杨忠云,李雪森. 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[J]. 现代肿瘤医学, 2023, 0(1): 17-24. DOI: 10.3969/j.issn.1672-4992.2023.01.003 |
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| 作者姓名： | 唐颖 李金玲 王科江 杨忠云 李雪森 |
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| 作者单位： | 西南医科大学基础医学院肿瘤研究所,四川 泸州 646000 |
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| 基金项目： | National Natural Science Foundation of China（No.81603154）;国家自然科学青年科学基金项目（编号:81603154） |
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| 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20 μmol/L和40 μmol/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40 μmol/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。
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| 关 键 词： | 肺癌 黄卡瓦胡椒素B EGFR-AKT信号通路 |
Flavokavin B promotes EGFR degradation and inhibits H1975 lung cancer cell proliferation and migration |
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| TANG Ying,LI Jinling,WANG Kejiang,YANG Zhongyun,LI Xuesen. Flavokavin B promotes EGFR degradation and inhibits H1975 lung cancer cell proliferation and migration[J]. Journal of Modern Oncology, 2023, 0(1): 17-24. DOI: 10.3969/j.issn.1672-4992.2023.01.003 |
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| Authors: | TANG Ying LI Jinling WANG Kejiang YANG Zhongyun LI Xuesen |
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| Affiliation: | Institute for Cancer Medicine,School of Basic Medical Sciences,Southwest Medical University,Sichuan Luzhou 646000,China. |
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| Abstract: | 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 20 μmol/L and 40 μmol/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/Ⅱ protein 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-Ⅱ protein 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. |
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| Keywords: | lung cancer Flavokavin B EGFR-AKT signaling pathway |
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