巨噬细胞刺激1受体抑制剂BMS777607对肺癌细胞增殖和凋亡的影响  

Effects of macrophage stimulating 1 receptor inhibitor BMS777607 on proliferation and apoptosis of lung cancer cells

作　　者：贾答淇 孔雯聪 苏荣健 杜晓媛 贺武斌 JIA Daqi;KONG Wencong;SU Rongjian;DU Xiaoyuan;HE Wubin(Department of Pathology,School of Basic Medical Science,Jinzhou Medical University,Jinzhou 121000,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4期739-744,I0002,共7页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81172048);辽宁省科技厅自然科学基金资助课题(2015020358)。

摘　　要：目的:探讨巨噬细胞刺激1受体(MST1R)抑制剂BMS777607对肺癌A549细胞增殖和凋亡的影响,阐明其可能的机制。方法:选择人肺癌A549细胞,不同浓度(1、5、10、15和20μmol·L^-1)BMS777607处理细胞48 h,同时设对照组,采用MTT法检测各组细胞增殖率;不同浓度(1、5、10、15和20μmol·L^-1)BMS777607处理A549细胞14 d,同时设对照组,采用克隆形成实验观察各组细胞克隆形成情况并检测各组细胞增殖率;不同浓度(10和20μmol·L^-1)BMS777607处理A549细胞24 h,同时设对照组,采用EdU掺入法检测各组细胞中EdU阳性细胞率;不同浓度(1、5、10、15和20μmol·L^-1)BMS777607处理A549细胞48 h,同时设对照组,采用流式细胞术检测各组细胞凋亡率;应用Western blotting法检测各组细胞中蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、细胞外调节蛋白激酶(ERK)、磷酸化ERK(p-ERK)、聚酰苷二磷酸核糖聚合酶(PARP)、活化型PARP(Cleaved-PARP)、含半胱氨酸的天冬氨酸蛋白水解酶9(Caspase 9)和活化型Caspase 9(Cleaved-Caspase 9)蛋白表达水平。结果:MTT实验,与对照组比较,不同浓度BMS777607组细胞增殖率明显降低(P<0.05或P<0.01),且呈浓度和时间依赖性。克隆形成实验,与对照组比较,10μmol·L^-1 BMS777607组细胞克隆形成数量明显减少,20μmol·L^-1 BMS777607组细胞几乎无克隆形成;与对照组比较,不同浓度BMS777607组细胞增殖率明显降低(P<0.05或P<0.01)。EdU掺入法实验,与对照组比较,10和20μmol·L^-1 BMS777607组细胞中EdU阳性细胞率均明显降低(P<0.05或P<0.01)。双染流式细胞术检测,与对照组比较,10、15和20μmol·L^-1 BMS777607组A549细胞凋亡率明显升高(P<0.05)。Western blotting法检测,与对照组比较,不同浓度BMS777607组细胞中p-AKT和p-ERK蛋白表达水平明显降低(P<0.05),Cleaved-PARP和Cleaved-Caspase 9蛋白表达水平明显升高(P<0.05)。结论:MST1R抑制剂BMS777607能够抑制肺癌A549细胞的增殖,并诱导其凋亡,其机制可能与�Objective:To investigate the effects of macrophage stimulating 1 receptor(MSTIR)inhibitor BMS777607 on the proliferation and apoptosis of lung cancer A549 cells,and to clarify their possible mechanisms.Methods:The A549 cells of human lung cancer were selected and treated with BMS777607 at different concentrations(1,5,10,15,and 20μmol·L^-1)for 48 h;at the same time,control group was set up.The proliferation rates of A549 cells in various groups were detected by MTT method.The A549 cells were treated with BMS777607 at different concentratio ns(1,5,10,15,and 20μmol·L^-1)for 14 d;at the same time,control group was set up.The colony formation of A549 cells in various groups were observed by colony formation experiment,and the proliferation rates of A549 cells were detected.The A549 cells were treated with different concentrations(10 and 20μmol·L^-1)of BMS777607 for 24 h;at the same time,control group was set up.The number of EdU positive cells in various groups were detected by EdU incorporation method.The A549 cells were treated with BMS777607 at different concentrations(1,5,10,15,20μmol·L^-1)for 48 h;at the same time,control group was set up.The apoptotic rates were detected by flow cytometry.Western blotting method was used to detect the expression levels of protein kinase B(AKT),phosphorylated AKT(p-AKT),extracellular regulatory protein kinase(ERK),phosphorylated ERK(p-ERK),polyglycoside diphosphate ribose polymerase(PARP),Cleaved-PARP,Caspase 9 and Cleaved-Caspase 9.Results:The MTT assay results showed that compared with control group,the proliferation rates of A54 cells in different concentrations of BMS777607 groups were decreased significantly(P<0.05 or P<0.01)in a concentration and time dependent manner.In colony formation experiment,compared with control group,the number of colony formation of A549 cells in 10μmol·L^-1 BMS777607 group was significantly decreased,and the colony formation almost was not found in 20μmol·L^-1 BMS777607 group;compared with control group,the colony formation rates in d

关 键 词：巨噬细胞刺激1受体 BMS777607 肺肿瘤 细胞凋亡 磷酸化蛋白激酶B 聚酰苷二磷酸核糖聚合酶 

分 类 号：R734.2[医药卫生—肿瘤]