基于GPER介导途径的二氢丹参酮Ⅰ抑制人乳腺癌MCF-7细胞增殖的分子机制研究  

Molecular mechanism of dihydrotanshinoneⅠon inhibiting proliferation of human breast cancer MCF-7 cells through GPER pathway

作　　者：程慧凯 赵丕文 胡周 CHENG Huikai;ZHAO Piwen;HU Zhou(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学生命科学学院,北京102488

出　　处：《上海中医药大学学报》2020年第2期54-59,共6页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81673764);北京中医药大学中央高校基本科研业务专项资金资助项目(2017-JYB-JS-001)。

摘　　要：目的:基于G蛋白偶联雌激素受体(GPER)途径探讨二氢丹参酮Ⅰ(DHⅠ)对人乳腺癌MCF-7细胞增殖的调控作用及其分子机制。方法:①将细胞分为对照组和DHⅠ不同浓度(0.1、0.2、0.3、0.4、0.5μg/ml)组,各组分别予以DMSO或相应浓度的DHⅠ干预。细胞培养48 h后,MTT法检测各组细胞增殖抑制率。②将细胞分为对照组、DHⅠ组、DHⅠ+激动剂组和DHⅠ+抑制剂组。先给予DHⅠ组、DHⅠ+激动剂组和DHⅠ+抑制剂组0.1μg/ml DHⅠ处理48 h,再向DHⅠ+激动剂组和DHⅠ+抑制剂组分别加入1μg/ml GPER激动剂G1和1μg/ml磷脂酰肌醇-3-激酶(PI3K)抑制剂LY294002干预,继续培养24 h后,Western blot检测GPER、PI3K、细胞周期蛋白D1(cyclinD1)、细胞周期蛋白依赖激酶2(CDK2)、蛋白激酶B(AKT)的蛋白表达。结果:①0.1~0.5μg/ml DHⅠ作用48 h后均可显著抑制MCF-7细胞的增殖,细胞增殖抑制率较对照组显著升高(P<0.01)。②0.1μg/ml DHⅠ作用48 h后,MCF-7细胞GPER、cyclinD1、CDK2、PI3K、AKT蛋白表达量较对照组明显降低(P<0.05,P<0.01)。③激动剂干预后,与DHⅠ组相比,DHⅠ+激动剂组细胞的GPER蛋白表达量明显升高(P<0.05),且PI3K、AKT、cyclinD1蛋白表达量显著增多(P<0.05,P<0.01)。④抑制剂干预后,与DHⅠ组相比,DHⅠ+抑制剂组cyclinD1蛋白表达水平进一步降低(P<0.05)。结论:DHⅠ可能通过GPER介导的PI3K/AKT途径抑制乳腺癌MCF-7细胞增殖。Objective:To investigate the regulatory effects of dihydrotanshinoneⅠ(DHⅠ)on the proliferation of human breast cancer MCF-7 cells,and discuss its molecular mechanism based on G protein-coupled estrogen receptor(GPER)pathway.Methods:①The cells were divided into the control group and DHⅠtreatment groups with different concentrations(0.1,0.2,0.3,0.4,0.5μg/ml).Each group was treated with DMSO or DHⅠat corresponding concentrations,respectively.After cell culture for 48 hours,the inhibition rate of cell proliferation in each group was detected by MTT assay.②The cells were divided into the control group,DHⅠgroup,DHⅠ+agonist group and DHⅠ+inhibitor group.The DHⅠgroup,DHⅠ+agonist group and DHⅠ+inhibitor group were treated with 0.1μg/ml DHⅠfor 48 hours,and then 1μg/ml G1(GPER agonist)and 1μg/ml LY294002[phosphatidylinositol-3-kinase(PI3 K)inhibitor]were added into the DHⅠ+agonist group and DHⅠ+inhibitor group,respectively.After cell culture for 24 hours,the protein expressions of GPER,PI3 K,cyclinD1,cyclin-dependent kinase 2(CDK2)and serine/threonine kinase(AKT)were detected by Western blot.Results:(1)The proliferation of MCF-7 cells was inhibited by DHⅠat 0.1-0.5μg/ml for 48 hours,and the inhibition rate of cell proliferation was significantly increased compared with the control group(P<0.01).(2)After treatment with DHⅠat 0.1μg/ml for 48 hours,the protein expressions of GPER,cyclinD1,CDK2,PI3 K and AKT in MCF-7 cells were significantly decreased compared with the control group(P<0.05,P<0.01).(3)After treatment with agonist,compared with DHⅠgroup,the expression of GPER protein was significantly increased in DHⅠ+agonist group,and the protein expressions of PI3 K,AKT and cyclinD1 were obviously increased(P<0.05,P<0.01).(4)After treatment with inhibitor,compared with DHⅠgroup,the expression level of cyclinD1 protein in DHⅠ+inhibitor group was further decreased(P<0.05).Conclusion:DHⅠmay inhibit the proliferation of breast cancer MCF-7 cells through GPERmediated PI3 K/AKT pathwa

关 键 词：二氢丹参酮Ⅰ 乳腺癌 细胞增殖 GPER PI3K/AKT MCF-7细胞 

分 类 号：R285[医药卫生—中药学]