石丹宁;崔丽霞;赵丕文;孙丽萍;陈梦;牛建昭;

• 1:北京中医药大学生命科学学院
• 2:北京中医药大学中医学院

摘要(Abstract)：

基于G蛋白偶联雌激素受体(GPER)及其介导的PI3K/AKT途径探讨隐丹参酮(cryptotanshinone,CPT)诱导人乳腺癌细胞凋亡的效应及其分子机制。选用核ER阴性、GPER阳性人乳腺癌SKBR-3细胞,采用Annexin V-FITC/PI双染法检测CPT对细胞凋亡率的影响,Western blot法检测CPT对凋亡效应蛋白caspase-3表达的影响; Western blot及免疫荧光技术检测CPT对GPER介导的PI3K/AKT通路上关键蛋白表达的调控效应;同时,为进一步明确GPER介导的PI3K/AKT信号通路在CPT诱导SKBR-3细胞凋亡中的作用,选用GPER特异性激动剂G1、特异性拮抗剂G15及PI3K特异性抑制剂LY294002进行干预。结果显示,5,10μmol·L~(-1)CPT作用48 h后,SKBR-3细胞的凋亡率上升为46. 1%,69. 0%(P<0. 01),caspase-3表达则随着CPT浓度增加而升高(P<0. 01)。PI3K,AKT,p-AKT蛋白表达均可被CPT抑制(P<0. 05或P<0. 01),G1与5μmol·L~(-1)CPT联合作用48 h后可见AKT与p-AKT的表达量进一步降低(P<0. 05),而G15与5μmol·L~(-1)CPT联合作用后AKT与p-AKT表达的下降被明显抑制(P<0. 05); 1μmol·L~(-1)LY294002干预下,可见AKT,p-AKT的表达受到了更为明显的抑制(P <0. 01)。AKT与p-AKT的荧光表达强度同样能够受到CPT抑制,G1,G15,LY294002的干预与Western blot结果一致(P <0. 05或P <0. 01)。此外,CPT对SKBR-3细胞中GPER表达具有一定的抑制效应(P<0. 01),加入G1可相对抑制CPT对GPER表达的下调作用;加入G15可使CPT对GPER表达的抑制作用进一步增强。综上,隐丹参酮可诱导人乳腺癌SKBR-3细胞凋亡,其分子途径可能与隐丹参酮对GPER表达及其介导的PI3K/AKT信号通路的调控作用有关。

关键词(KeyWords)： 隐丹参酮;GPER;乳腺癌;SKBR-3;PI3K/AKT;细胞凋亡;植物雌激素

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673764);; 教育部&国家外国专家局“北京中医药大学中西医结合学科创新引智基地”项目(B07007);; 北京中医药大学研究生自主课题(2018-JYB-XS100);北京中医药大学中央高校基本科研业务费专项(2017-JYB-JS-001)

作者(Author): 石丹宁;崔丽霞;赵丕文;孙丽萍;陈梦;牛建昭;

Email:

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