MST1R抑制剂BMS-777607对乳腺癌MCF-7细胞增殖的抑制和凋亡诱导作用  

Inhibitory effect and apoptosis induction of MST1R inhibitor BMS-777607 on proliferation of breast cancer MCF-7 cells

作　　者：孔雯聪 贺武斌 苏荣健 贾答淇 谷艳娇 王月 杜晓媛 KONG Wencong;HE Wubin;SU Rongjian;JIA Daqi;GU Yanjiao;WANG Yue;DU Xiaoyuan(Department of Pathology, School of Basic Medical Sciences, Jinzhou Medical University, Jinzhou 121000, China;First Affiliated Hospital, Jinzhou Medical University, Jinzhou 121001, China;Department of Cell Biology, School of Basic Medical Sciences, Jinzhou Medical University, Jinzhou 121000, China)

机构地区：[1]锦州医科大学基础医学院病理学教研室,辽宁锦州121000 [2]锦州医科大学附属第一医院,辽宁锦州121001 [3]锦州医科大学基础医学院细胞生物学教研室,辽宁锦州121000

出　　处：《吉林大学学报（医学版）》2020年第3期451-457,共7页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81172048);辽宁省教育厅自然科学基金资助课题(2015020326)。

摘　　要：目的:探讨巨噬细胞刺激1受体(MST1R)抑制剂BMS-777607对乳腺癌MCF-7细胞增殖和凋亡的影响,并阐明其作用机制。方法:采用不同浓度BMS-777607处理乳腺癌MCF-7细胞,将细胞分为对照组(0μmol·L-1 BMS-777607组)和0.5、1.0、2.0、5.0、10.0、15.0及20.0μmol·L-1 BMS-777607组。采用MTT法检测各组MCF-7细胞增殖率,克隆形成实验检测各组MCF-7细胞存活率,EDU成像和EDU流式细胞术检测各组MCF-7细胞增殖率,Hoechst33342染色法检测各组MCF-7细胞凋亡形态表现,流式细胞术检测各组MCF-7细胞凋亡率,Western blotting法检测各组MCF-7细胞中ERK、p-ERK、Akt、p-Akt、PARP、Cleaved PARP、Bax、Caspase-3、Cleaved Caspase-3、Caspase-9和Cleaved Caspase-9蛋白表达水平。结果:MTT检测,与对照组比较,5和10μmol·L-1BMS-777607组MCF-7细胞增殖率升高(P<0.05或P<0.01)。克隆形成实验,与对照组比较,5和20μmol·L-1BMS-777607组MCF-7细胞存活率升高(P<0.05或P<0.01)。EDU掺入法检测,与对照组比较,10和20μmol·L-1BMS-777607组MCF-7细胞增殖率升高(P<0.05或P<0.01)。Hoechst33342荧光染色,对照组MCF-7细胞核淡染,10μmol·L-1BMS-777607组MCF-7细胞核少部分浓染、明亮,20μmol·L-1BMS-777607组MCF-7细胞核大部分浓染,细胞核染色质固缩、明亮。双染流式细胞术检测,与对照组比较,10和20μmol·L-1BMS-777607组MCF-7细胞凋亡率降低(P<0.05)。Western blotting法检测,与对照组比较,10、15和20μmol·L-1BMS-777607组MCF-7细胞中p-ERK和p-Akt蛋白表达水平明显降低(P<0.05或P<0.01),PARP、Cleaved PARP、Bax、Cleaved Caspase-3及Cleaved Caspase-9蛋白表达水平明显升高(P<0.05或P<0.01)。结论:MST1R抑制剂BMS-777607能够抑制乳腺癌MCF-7细胞增殖并诱导其凋亡,其作用机制与抑制p-ERK和p-Akt表达和促进Cleaved PARP、Bax、Cleaved Caspase-9及Cleaved Caspase-3表达有关。Objective:To investigate the effects of macrophage stimulating 1 receptor(MST1R)inhibitor BMS-777607 on the proliferation and apoptosis of the breast cancer MCF-7 cells,and to elucidate the mechanisms.Methods:The breast cancer MCF-7 cells treated with different concentrations of BMS777607 were divided into control group(0μmol·L-1 BMS-777607 group)and 0.5,1.0,2.0,5.0,10.0,15.0,and 20.0μmol·L-1 BMS-777607 groups.MTT method was used to detect the proliferation rates of the MCF-7 cells in various groups,and clone formation assay was used to detect the survival rates of the MCF-7 cells in various groups;EDU imaging and EDU flow cytometry methods were used to detect the proliferation rates of the MCF-7 cells in various groups,and Hoechst33342 staining was used to detect the apoptotic morphology of the MCF-7 cells in various groups;flow cytometry was used to detect the apoptotic rates of the MCF-7 cells in various groups,and Western blotting method was used to detect the expression levels of ERK,p-ERK,Akt,p-Akt,PARP,Cleaved PARP,Bax,Caspase-3,Cleaved Caspase-3,Caspase-9 and Cleaved Caspase-9 proteins in the MCF-7 cells in various groups.Results:The MTT results showed that compared with control group,the proliferation rates of the MCF-7 cells in 5 and 10μmol·L-1BMS-777607 groups were increased(P<0.05 or P<0.01).The clone formation assay results showed that compared with control group,the survival rates of the MCF-7 cells in 5 and 20μmol·L-1BMS-777607 groups were increased(P<0.05 or P<0.01).The EDU incorporation results showed that compared with control group,the proliferation rates of the MCF-7 cells in 10 and 20μmol·L-1BMS-777607 groups were increased(P<0.05 or P<0.01).The Hoechst33342 fluorescence staining results showed that the MCF-7 cells in control group showed the light staining,a small number of MCF-7 cells in 10μmol·L-1BMS-777607 group showed the bright nuclei,and most of the MCF-7 cells in 20μmol·L-1BMS-777607 group showed the bright nuclei.The flow cytometry results showed that compared with cont

关 键 词：BMS-777607 乳腺肿瘤 细胞增殖 细胞凋亡 巨噬细胞刺激1受体 

分 类 号：R737.9[医药卫生—肿瘤]