三叶青总黄酮通过降低糖酵解增强NSCLC耐药细胞对吉非替尼敏感性的实验研究
Total Flavonoids of Tetrastigma Hemsleyanum Enhanced the Eensitivity of NSCLC Resistant Cells to Gefitinib by Decreasing Glycolysis
何佳奇 1陈瑞琳 2解伟 3李雪健 3余陈欢4
作者信息
- 1. 浙江省立同德医院 浙江 杭州 310012;浙江省中医药研究院 浙江 杭州 310007
- 2. 浙江中医药大学附属第一医院 浙江 杭州 310006
- 3. 杭州医学院 浙江 杭州 310013
- 4. 中国科学院杭州医学研究所 浙江 杭州 310018
- 折叠
摘要
目的:观察三叶青总黄酮对非小细胞肺癌(non-small cell lung cancer,NSCLC)吉非替尼(Gefi-tinib,GEF)耐药细胞增殖的影响,并通过糖酵解途径探讨其可能的作用机制.方法:体外培养NSCLC细胞A549和H1975及吉非替尼耐药细胞A549/GR和H1975/GR,CCK-8法检测三叶青总黄酮单用以及与吉非替尼联用对每组细胞增殖的影响;ELISA法检测每组细胞葡萄糖消耗量、ATP释放量、乳酸及丙酮酸生成量;Western blot检测每组细胞糖酵解关键酶葡萄糖转运蛋白1(GLUTl)、乳酸脱氢酶A(LDHA)、己糖激酶2(HK2)、M2型丙酮酸激酶(PKM2)蛋白表达水平.结果:三叶青总黄酮能显著抑制耐药细胞的葡萄糖摄取,减少ATP、乳酸及丙酮酸产生,增加耐药细胞对吉非替尼的敏感性,进而抑制吉非替尼耐药NSCLC细胞株的增殖,能显著降低A549/GR和H1975/GR细胞GLUTl、LDHA、HK2、PKM2蛋白表达水平.结论:三叶青总黄酮可增强NSCLC耐药细胞对吉非替尼的敏感性,其机理可能与抑制糖酵解相关.
Abstract
Objective:To observe the effects of total flavonoids of Tetrastigma hemsleyanum on the prolifera-tion of Gefitinib(GEF)resistant cells in non-small cell lung cancer(NSCLC),and explore its possible mechanism through glycolysis.Methods:NSCLC cells A549 and H1975 and gefitinib resistant cells A549/GR and H1975/GR were cultured in vitro.CCK-8 method was used to detect the effects of Tetrastigma hemsleya-num alone or in combination with gefitinib on cell proliferation in each group.ELISA method was used to detect glucose consumption,ATP release,lactic acid and pyruvate production.Westen blot was used to detect the protein expression levels of glucose transporter 1(GLUTl),lactate dehydrogenase A(LDHA),hexokinase 2(HK2)and M2 pyruvate kinase(PKM2).Results:The total flavonoids of Tetrastigma hemsleyanum signifi-cantly inhibited the glucose uptake,decreased the production of ATP,lactic acid and pyruvate production in drug-resistant cells,increased the sensitivity of drug-resistant cells to gefitinib,and thereby inhibited the pro-liferation of gefitinib resistant NSCLC cell lines.It significantly decreased the proteins expression levels of GLUTl,LDHA,HK2,and PKM2 in A549/GR and H1975/GR cells.Conclusion:The total flavonoids of Tetra-stigma hemsleyanum can enhance the sensitivity of NSCLC resistant cells to gefitinib,and its mechanism may be related to the inhibition of glycolysis.
关键词
三叶青总黄酮/NSCLC/吉非替尼/耐药/糖酵解Key words
Total flavonoids of Tetrastigma hemsleyanum/NSCLC/gefitinib/drug resistance/glycolysis引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据