吉西他滨靶向HGF/cMET通路对非小细胞肺癌A549细胞迁移和侵袭的影响  

Effect of gemcitabine on migration and invasion of non-small lung cancer A549 cells via targeting HGF/cMET pathway

作　　者：郑琪[1] 廖子君[1] 马婕群 张彦兵[1] 李索妮[1] 李倩[1,2] 白杰[1,2] ZHENG Qi;LIAO Zijun;MA Jiequn;ZHANG Yanbing;LI Suoni;LI Qian;BAI Jie(First Department of Medical Oncology,Affiliated Shaanxi Provincial Tumor Hospital,College of Medicine,Xi’an Jiaotong University,Xi’an 710061,China;Xi’an Medical University)

机构地区：[1]西安交通大学医学院附属陕西省肿瘤医院内一科,西安710061 [2]西安医学院

出　　处：《山西医科大学学报》2020年第4期275-282,共8页Journal of Shanxi Medical University

基　　金：西安市科技计划项目(2017113SF/YX007(4))。

摘　　要：目的研究吉西他滨对非小细胞肺癌A549细胞迁移和侵袭的影响和机制.方法A549细胞加入不同浓度的吉西他滨(0,25,50,75 nmol/L)作用24,48,72,96 h,MTT法检测细胞增殖活力.将50 nmol/L的吉西他滨加入A549细胞,流式细胞仪分析吉西他滨对细胞周期和细胞凋亡的影响.A549细胞分为对照组、吉西他滨50 nmol/L组、肝细胞生长因子(50μg/L)组和吉西他滨(50 nmol/L)联合肝细胞生长因子(50μg/L)组,各组分别加入不同的药物,Transwell法检测各组细胞迁移能力和侵袭能力,Western blot法检测各组细胞p-cMET蛋白和cMET蛋白表达水平.结果A549细胞加入不同浓度的吉西他滨分别处理24,48,72,96 h,与0 nmol/L相比,25 nmol/L吉西他滨处理后细胞增殖活力无明显变化(P>0.05),50 nmol/L和75 nmol/L吉西他滨处理后的细胞增殖活力明显下降(P<0.05),但50 nmol/L与75 nmol/L吉西他滨处理后细胞增殖活力无明显差异(P>0.05).与对照组相比,50 nmol/L吉西他滨组的G1/G0期细胞比例轻度下降(P>0.05),而S期细胞比例明显上升(P<0.05),G2期细胞比例轻度下降(P>0.05).与对照组相比,50 nmol/L吉西他滨组无论是早期凋亡细胞比例还是晚期凋亡细胞比例,均明显升高(P<0.05).与对照组相比,吉西他滨组细胞迁移和侵袭能力明显下降(P<0.05),肝细胞生长因子组细胞迁移和侵袭能力明显升高(P<0.05),吉西他滨联合肝细胞生长因子联合组细胞迁移和侵袭能力较单独吉西他滨组升高,低于单独加入肝细胞生长因子组,与对照组无明显差异(P>0.05).与对照组相比,吉西他滨组p-cMET蛋白水平明显下降(0.4512±0.1109 vs 0.1076±0.0082,P<0.05),肝细胞生长因子组p-cMET蛋白水平显著升高(0.4512±0.1109 vs 0.7835±0.1206,P<0.05).而与对照组相比,吉西他滨组和肝细胞生长因子组的cMET蛋白表达水平无明显变化(P>0.05).结论吉西他滨可通过下调HGF/cMET通路抑制肺癌A549细胞的侵袭和迁移能力.Objective To explore the effects of gemcitabine on the migration and invasion of A549 cells and its mechanism.Methods A549 cells were treated with 0,25,50,75 nmol/L gemcitabine for 24,48,72,96 h,respectively,and then MTT assay was used to detect the cell proliferation.Then A549 cells were added with 50 nmol/L gemcitabine,and the cell cycle and the apoptosis of A549 cells were examined with flow cytometer.A549 cells were divided into control group,gemcitabine(50 nmol/L)group,hepatocyte growth factor(50μg/L)gourp and gemcitabine(50 nmol/L)in combination with hepatocyte growth factor(50μg/L)group.The migration and invasion abilities of A549 cells were tested with Transwell method,and p-cMET and cMET protein levels were examined by Western blot method.Results Compared with 0 nmol/L group,the cell proliferation ability in 25 nmol/L group showed no significant difference(P>0.05),and the cell proliferation abilities in 50 nmol/L and 75 nmol/L groups decreased significantly(P<0.05),but there was no significant difference between 50 nmol/L group and 75 nmol/L group(P>0.05).Compared with control group,the numbers of cells dropped a little in G1/G0 and G2 phases in 50 nmol/L gemcitabine group(P>0.05),but increased significantly in S phase(P<0.05),and both early and late apoptotic rates of cells in 50 nmol/L gemcitabine group increased compared with control group(P<0.05).Compared with control group,the cell migration and invasion abilities were decreased significantly in gemcitabine group(P<0.05),but increased in hepatocyte growth factor group(P<0.05).The abilities in gemcitabine+hepatocyte growth factor group were higher than those in gemcitabine group(P<0.05),lower than those of hepatocyte growth factor group(P<0.05),but the abilities showed no difference compared with control group(P>0.05).Compared with control group,the p-cMET protein expression level was dropped in gemcitabine group(0.4512±0.1109 vs 0.1076±0.0082,P<0.05),but elevated in hepatocyte growth factor group(0.4512±0.1109 vs 0.7835±0.1206,P<0.05),but the

关 键 词：吉西他滨 非小细胞肺癌 细胞迁移 细胞侵袭 

分 类 号：R734.2[医药卫生—肿瘤]