三羧基取代夹心型MnⅢAs2W18对非小细胞肺癌细胞株A549上皮-间充质转化的影响及其机制  

Impact and mechanism of PMs-3 to epithelial mesenchymal transformation of non-small cell lung cancer cell line A549

作　　者：陈子豪 李仙 贾静 李萍 姜英虹 句红萍 Chen Zihao;Li Xian;Jia Jing;Li Ping;Jiang Yinghong;Ju Hongping(Graduate School of Hebei Medical University,Shijiazhuang 050011,China;School of Medicine,Kunming University,Kunming 650214,China)

机构地区：[1]河北医科大学研究生学院,石家庄050117 [2]昆明学院医学院,650214

出　　处：《中华实验外科杂志》2019年第9期1536-1539,共4页Chinese Journal of Experimental Surgery

基　　金：云南省高校联合面上项目(2017FH001-083).

摘　　要：目的探讨三羧基取代夹心型MnⅢAs2W18(PMs-3)对非小细胞肺癌(NSCLC)细胞上皮-间充质转化(EMT)的影响,并分析其分子机制。方法应用本课题组前期合成的PMs-3纳米复合结构以0、20、40、60、80、100、120 nmol/L浓度作用人NSCLC细胞株A549及人正常肺上皮细胞株BEAS-2B 48 h,噻唑蓝(MTT)实验检测PMs-3对细胞的抑制作用;划痕实验及Transwell小室实验检测PMs-3作用后A549细胞的迁移侵袭能力的变化;实时定量反转录聚合酶链反应(RT-qPCR)及蛋白质印迹法(Western blot)检测PMs-3作用后A549细胞EMT相关基因的变化。结果MTT结果显示,PMs-3对A549细胞的活性具有抑制作用,抑制率:0 nmol/L为(3.203±1.222)%、20 nmol/L为(11.958±4.000)%、40 nmol/L为(25.348±7.710)%、60 nmol/L为(34.775±9.155)%、80 nmol/L为(41.163±10.988)%、100 nmol/L为(50.430±9.743)%、120 nmol/L为(51.895±6.161)%,呈剂量依赖性(F=35.451,P<0.01);100 nmol/L的PMs-3对BEAS-2B细胞的抑制率[(24.103±5.755)%]低于对A549细胞(t=5.699,P<0.01);PMs-3作用后A549细胞的迁移率[(34.545±7.888)%]明显低于对照组[(77.408±5.442)%](t=-10.956,P<0.01);PMs-3作用后A549细胞侵袭能力明显减弱,实验组穿膜细胞为(125.333±25.137),对照组为(200.667±19.967,t=-5.748,P<0.01);细胞中EMT相关基因的mRNA和蛋白表达发生变化:E-钙黏蛋白(E-cadherin)表达明显增强,N-钙黏蛋白(N-cadherin)、Snail表达明显减弱(t=-5.753、-10.538、5.583、5.771、6.660、4.120,P<0.05),波形蛋白(Vimentin)表达无明显变化(t=-0.323、1.179,P>0.05)。结论PMs-3可通过抑制NSCLC细胞EMT而发挥作用。Objective To explore Impact and mechanism of PMs-3 to epithelial mesenchymal transformation(EMT)of non-small cell lung cancer(NSCLC)cell line A549.Methods Nanocomposite structure synthetized by our team was utilized on human NSCLC cell line A549 and human normal pulmonary epithelial cell line BEAS-2B with concentrations of 0,20,40,60,80,100,120 nmol/L for 48 h.Methyl thiazol tetrazolium(MTT)assay was employed to detect the effect of PMs-3 on A549 cells.Wound assay and Transwell chamber assay were used to test the migration and invasion of PMs-3 on A549 cells.EMT related genes were detected with real time PCR(RT-qPCR)and Western blotting.Results Results of MTT assay detected that PMs-3 could inhibit activity of A549 cells significantly,and inhibition rate was(3.203±1.222)%(0 nmol/L),(11.958±4.000)%(20 nmol/L),(25.348±7.710)%(40 nmol/L),(34.775±9.155)%(60 nmol/L),(41.163±10.988)%(80 nmol/L),(50.430±9.743)%(100 nmol/L),(51.895±6.161)%(120 nmol/L),which showed a dose-dependent manner(F=35.451,P<0.01).Inhibition rate of BEAS-2B cells treated with PMs-3(100 nmol/L)was(24.103±5.755)%,which was lower than that of A549 cells(t=5.699,P<0.01).Migrationrate of A549 cells in PMs-3 group was(34.545±7.888)%,which was lower than in control group(77.408±5.442)(t=-10.956,P<0.01).Cells through the Transwell chamber in PMs-3 group was 125.333±25.137),which was less than in control group(200.667±19.967)(t=-5.748,P<0.01).mRNA and proteins of EMT related genes varied after PMs-3 treated as following:expression of E-cadherin increased significantly in PMs-3 group compared with control group,while expression of N-cadherin,Snail decreased significantly in PMs-3 group compared with control group(t=-5.753,-10.538,5.583,5.771,6.660,4.120,P<0.05);no significant variation of Vimentin was verified between PMs-3 group and control group(t=-0.323,1.179,P>0.05).Conclusion PMs-3 could inhibit NSCLC cells via regulating EMT.

关 键 词：非小细胞肺癌 PMs-3纳米复合结构 上皮-间充质转化 药物治疗 

分 类 号：R734.2[医药卫生—肿瘤]