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

作　　者：何佳奇 陈瑞琳[3] 解伟 李雪健 余陈欢 HE Jiaqi;CHEN Ruilin;XIE Wei;LI Xuejian;YU Chenhuan(Tongde Hospital of Zhejiang Province,Hangzhou,310012;Zhejiang Academy of Traditional Chinese Medicine,Hangzhou,310007;The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou,310006;Hangzhou Medical college,Hangzhou,310013;Hangzhou Institute of Medicine,Chinese Academy of Sciences,Hangzhou,310018)

机构地区：[1]浙江省立同德医院,浙江杭州310012 [2]浙江省中医药研究院,浙江杭州310007 [3]浙江中医药大学附属第一医院,浙江杭州310006 [4]杭州医学院,浙江杭州310013 [5]中国科学院杭州医学研究所,浙江杭州310018

出　　处：《浙江中医杂志》2024年第12期1035-1039,共5页Zhejiang Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目三叶青黄酮类成分调控节律基因-糖酵解途径逆转NSCLC获得性EGFR-TKIs耐药的机制研究,编号:82104526;浙江省医药卫生科技计划项目三叶青异牡荆素逆转非小细胞肺癌EGFR-TKIs耐药的糖酵解节律调控机制研究,编号:2022KY917;第七批全国老中医药专家学术经验继承人项目,编号:国中医药人教函〔2022〕76号;全国中药特色技术传承人才培训项目,编号:国中医药人教函〔2023〕96号。

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。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 

分 类 号：R285[医药卫生—中药学]