重楼皂苷在miR-125a-5p靶向调控GAB2诱导乳腺癌细胞增殖和凋亡中的作用及机制研究  

Proliferation and apoptosis of breast cancer cells induced by rhizoma paridis saponins through miR-125a-5p targeting GAB2

作　　者：吕世文 朱亚兰[2] 王婧婧 LYU Shiwen;ZHU Yalan;WANG Jingjing(Department of Pharmacy,Jinhua Maternal&Child Health Care Hospital,Jinhua 321000,China;不详)

机构地区：[1]金华市妇幼保健院药学部,321000 [2]金华市中心医院药学部

出　　处：《浙江医学》2023年第20期2129-2135,2162,共8页Zhejiang Medical Journal

基　　金：金华市公益性技术应用研究项目(2022-04-191)。

摘　　要：目的探讨重楼皂苷(RPS)在miR-125a-5p靶向调控Grb2相关结合蛋白2(GAB2)诱导乳腺癌细胞增殖和凋亡中的作用及机制。方法取人乳腺癌细胞系MCF-7和人正常乳腺上皮细胞系MCF-10A,采用qRT-PCR、Western blot法分别检测并比较两者miR-125a-5p、GAB2表达水平。再将MCF-7细胞按随机数字表法分成6组:RPS 0μg/mL组、RPS 80μg/mL组、RPS 80μg/mL+阴性抗体组、RPS 80μg/mL+anti-miR-125a-5p组、RPS 80μg/mL+anti-miR-125a-5p+阴性干扰组和RPS 80μg/mL+anti-miR-125a-5p+si-GAB2组,每组设3个复孔;培养48 h后检测并比较miR-125a-5p、GAB2、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达水平以及细胞增殖率、细胞凋亡率。将GAB2与空白质粒和过表达miR-125a-5p质粒共转染到MCF-7细胞中,采用荧光素酶Promega检测法检测miR-125a-5p与GAB2的靶向作用。结果与MCF-10A细胞比较,MCF-7细胞中miR-125a-5p表达水平较低(P<0.05),GAB2表达水平较高(P<0.05)。与RPS 0μg/mL组比较,RPS 80μg/mL组和RPS 80μg/mL+阴性抗体组miR-125a-5p、Bax表达水平和细胞凋亡率均明显升高(均P<0.05),细胞增殖率和Bcl-2表达水平均明显降低(均P<0.05);RPS 80μg/mL组和RPS 80μg/mL+anti-miR-125a-5p+阴性干扰组GAB2表达水平均明显降低(均P<0.05)。与RPS 80μg/mL组、RPS 80μg/mL+阴性抗体组比较,RPS 80μg/mL+anti-miR-125a-5p组miR-125a-5p表达水平均明显降低(均P<0.05);RPS 80μg/mL+anti-miR-125a-5p组和RPS 80μg/mL+anti-miR-125a-5p+阴性干扰组细胞增殖率和Bcl-2表达水平均明显升高(均P<0.05),细胞凋亡率和Bax表达水平均明显降低(均P<0.05)。与RPS 80μg/mL组、RPS 80μg/mL+anti-miR-125a-5p+阴性干扰组比较,RPS 80μg/mL+anti-miR-125a-5p+si-GAB2组GAB2表达水平均明显降低(均P<0.05)。与RPS 80μg/mL+anti-miR-125a-5p组、RPS 80μg/mL+anti-miR-125a-5p+阴性干扰组比较,RPS 80μg/mL+anti-miR-125a-5p+si-GAB2组细胞增殖率和Bcl-2表达水平均明显降低(均P<0.05),细胞凋亡率和Bax表达水平均明显升高(�Objective To investigate the mechanism of rhizoma paridis saponins(RPS)acting on the proliferation and apoptosis of breast cancer cells induced by miR-125a-5p targeting Grb2 related binding protein 2(GAB2).Methods MCF-7 human breast cancer cell and MCF-10A human normal breast epithelial cell were chosed,and qRT-PCR and Western blot were used to detect expression levels of miR-125a-5p and GAB2.Then MCF-7 cells were randomly divided into 6 groups:RPS 0μg/mL group,RPS 80μg/mL group,RPS 80μg/mL+negative antibody group,RPS 80μg/mL+anti-miR-125a-5p group,RPS 80μg/mL+anti-miR-125a-5p+negative interference group and RPS 80μg/mL+anti-miR-125a-5p+si�GAB2 group,with 3 holes being set for each group.The expression levels of miR-125a-5p,GAB2,B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax),and proliferation and apoptosis rates were detected and compared after 48 hours of cultivation.GAB2 was co-transfected into MCF-7 cells with blank plasmid and overexpressed miR-125a-5p plasmid,then the targeting effect of miR-125a-5p on GAB2 was detected using Promega dual Luciferase system assay kit.Results Compared with MCF-10A,MCF-7 showed significantly lower expression of miR-125a-5p and higher expression of GAB2(both P<0.05).Compared with RPS 0μg/mL group,the expressions of miR-125a-5p and Bax and cell apoptosis rate were significantly higher in RPS 80μg/mL group and RPS 80μg/mL+negative antibody group,while their cell proliferation rate and Bcl-2 level were significantly decreased(all P<0.05).The GAB2 level in RPS 80μg/mL group and RPS 80μg/mL+anti-miR-125a-5p+negative interference group was significantly decreased,too(all P<0.05).The miR-125a-5p level in RPS 80μg/mL+anti-miR-125a-5p group was significantly less than RPS 80μg/mL group and RPS 80μg/mL+negative antibody group(both P<0.05).The cell proliferation rate and Bcl-2 level were significantly higher in RPS 80μg/mL+anti-miR-125a-5p group and RPS 80μg/mL+anti-miR-125a-5p+negative interference group(all P<0.05),and cell apoptosis rate and Bax level were

关 键 词：重楼皂苷 乳腺癌 miR-125a-5p Grb2相关结合蛋白2 增殖 凋亡 作用机制 

分 类 号：R285[医药卫生—中药学]