摘要
目的蛇葡萄素是存在于植物藤茶中含量非常高的黄酮类化合物,本研究主要探讨抑制Akt-mTOR/PKM2介导的糖酵解在蛇葡萄素诱导乳腺癌细胞增殖抑制中的作用。方法不同浓度蛇葡萄素(20、40、60和80μmol/L)分别处理MDA-MB-231、MCF-7人乳腺癌细胞24h后,CCK-8检测乳腺癌细胞活力,2-NBDG法检测细胞糖摄取能力,比色法检测细胞培养液中乳酸含量,Western blot检测相关分子蛋白的表达。结果蛇葡萄素可显著抑制乳腺癌细胞的增殖(P<0.05),且随着蛇葡萄素处理剂量的增加,乳腺癌细胞增殖抑制增强(P<0.05),提示蛇葡萄素诱导乳腺癌细胞增殖抑制作用存在明显的剂量效应关系。进一步研究发现,蛇葡萄素可剂量依赖性的降低乳腺癌细胞的糖摄取能力(P<0.05),并显著减少乳腺癌细胞乳酸生成(P<0.05)。Western blot检测发现,蛇葡萄素在显著下调Akt-mTOR信号通路中p-Akt、p-mTOR、p-p70S6K等蛋白表达水平的同时下调与糖酵解相关分子GLUTI、LDHA、PKM2的蛋白表达水平。预先给予Akt-mTOR信号通路诱导剂IGF-1,以上作用均可被显著削弱。结论蛇葡萄素可抑制乳腺癌增殖,该作用与其抑制Akt-mTOR信号通路、进而抑制乳腺癌细胞糖酵解有关,本研究结果可为蛇葡萄素抗肿瘤作用机制提供了一种新的解释。
Objective To explore whether AMP,a flavonol mainly found in Ampelopsis grossedentata,induces cell growth inhibition by suppressing glycolysis through Akt-mTOR/PKM2 pathway in breast cancer cells.Methods After MCF-7 and MDA-MB-231 breast cancer cells treated with 20,40,60 and 80μmol/L AMP for 24h,cell viability was assessed by CCK-8 assay,glucose uptake assay was performed using 2-NBDG,lactate levels were measured using the colorimetric lactate assay kit and the protein expressions were assessed by Western blot.Results AMP treatment could inhibit cell viability in MCF-7 and MDA-MB-231 breast cancer cells in dose-dependent manners.The further experiments exhibited that AMP successfully blocked cell glycolysis by inhibiting the level of glucose uptake and the production of lactic acid,decreased the level of glycolysis-related proteins pyruvate kinase M2(PKM2),glucose transporter1(GLUT1)and lactate dehydrogenase A(LDHA),and also inactivated the Akt-mTOR pathway by down-regulation of p-Akt、p-mTOR、p-p70S6K expressions.However,Akt-mTOR pathway inducer IGF-1 treatment significantly attenuated AMP-induced above mentioned results.Conclusion AMP suppressed breast cancer cell growth and glycolysis through inhibiting Akt-mTOR/PKM2 pathway,and may provide a potential therapeutic target for breast cancer treatment.
作者
周永
郑金英
徐小华
金小玲
ZHOU Yong;ZHENG Jin-ying;XU Xiao-hua;JIN Xiao-ling(Department of Clinic Nutrition,People s Hospital of Chongqing Banan District,Chongqing 401320,China)
出处
《肿瘤代谢与营养电子杂志》
2019年第2期236-241,共6页
Electronic Journal of Metabolism and Nutrition of Cancer
基金
重庆市基础科学与前沿技术研究项目(CSTC2017jcyjAX0305)
陆军军医大学电磁辐射医学防护教育部重点实验室开放课题(2017DCKF002)
