张莹,金爱,王旭东
ZHANG Ying,JIN Ai,WANG Xudong

• 1:贵州医科大学基础医学院生理学教研室

摘要(Abstract)：

目的:探讨17β-雌二醇素(E2)对人三阴乳腺癌(TNBC)细胞增殖和凋亡的影响及其作用机制。方法:以人源乳腺癌细胞系MDA-MB-231及468细胞为模型细胞,实验分为二甲基亚砜组(DMSO组)、E2组、E2联合钙蛋白酶抑制剂(E2联合Cal)组及E2联合钙蛋白酶抑制剂Ⅲ组(E2联合CIⅢ组),采用噻唑蓝(MTT)实验和平板克隆实验检测细胞增殖和克隆能力,采用V-FITC/PI荧光凋亡试剂盒检测细胞程序性死亡,采用蛋白印迹法检测细胞蛋白表达水平。结果:E2组两种细胞的增殖率、菌落形成率均显著高于DMSO组(P<0.01,或P<0.05),凋亡强度显著低于DMSO组,YAP蛋白表达显著高于DMSO组(P<0.05);E2联合Cal组和E2联合CIⅢ组中两种细胞的增殖率、细胞菌落形成率均显著低于E2组(P<0.05,或P<0.01),荧光凋亡强度显著高于E2组,YAP蛋白表达均显著高于E2组(P<0.05,或P<0.01)。结论:E2具有诱导模型细胞增殖、抵抗细胞凋亡的能力,Cal和CIⅢ可以阻断E2诱导的上述生物学效应,提示E2可能通过CANP-YAP信号通路促进肿瘤细胞的恶性生物学行为。
Objective: To investigate the effect of 17β-E2 on proliferation and apoptosis of TNBC and its mechanism.Methods:Human breast cancer cell lines MDA-MB-231 and 468 were used as model cells.The experimental groups were DMSO group,E2 group,E2+calpeptin(E2+Cal) group and E2+calpain inhibitor III(E2+CI III) group.MTT and plate cloning experiments were used to investigate cell proliferation and cloning ability; V-FITC/PI fluorescent apoptosis kit was used to detect programmed cell death; Western blot assay was used to detect the expression level of cell protein.Results:The proliferation rate and colony formation rate of two kinds of cells in E2 group were higher than that in DMSO group(P<0.01 or P<0.05),the apoptotic intensity in E2 group was lower than that in DMSO group,the expression of YAP protein in E2 group was higher than that in DMSO group(P<0.05); the proliferation rate and the colony formation rate of the two kinds of cells in E2 combined with Cal group and E2 combined with CIIII group was lower than that of E2 group(P<0.05 or P<0.01).The fluorescent apoptotic intensity of two kinds of cells in E2+Cal group and E2+CIIII group was significantly higher than that in E2 group,YAP protein expression was also higher than that of E2 group(P<0.05,or P<0.01).Conclusions:E2 could induce model cell proliferation,resist apotosis,Cal and CI III could inhibit previously mentioned biological effect induced by E2; indicating E2 could promote tumor cell malignant activities by CANP-YAP signal channel.

关键词(KeyWords)： 乳腺肿瘤;雌二醇;细胞增殖;细胞凋亡;钙蛋白酶;YES相关蛋白
breast neoplasms;estrogen;cell proliferation;apoptosis;calpain;YES-associated protein

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31660345,31360252)

作者(Author): 张莹,金爱,王旭东
ZHANG Ying,JIN Ai,WANG Xudong

DOI: 10.19367/j.cnki.1000-2707.2019.08.004

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