LAMC2对胃癌细胞侵袭、迁移的作用及其机制研究  

Effect of LAMC2 on invasion and migration of gastric cancer cells and its mechanism.WANG Qing-yun

作　　者：汪清云 王文博[1] 郭卫[2] 夏泠 WANG Qing-yun;WANG Wen-bo;GUO Wei;XIA Ling(Department of Medical Oncology and Radiotherapy,Abdominal Tumor Division,Zhongnan Hospital of Wuhan University,Wuhan 430071,Hubei,CHINA;Department of Pathology,Wuhan University TaiKang Medical school(School of Basic Medical Sciences),Wuhan 430071,Hubei,CHINA)

机构地区：[1]武汉大学中南医院腹部肿瘤放化疗科,湖北武汉430071 [2]武汉大学泰康医学院(基础医学院)病理教研室,湖北武汉430071

出　　处：《海南医学》2023年第6期761-768,共8页Hainan Medical Journal

摘　　要：目的探讨层黏连蛋白γ_(2)(LAMC2)对胃癌细胞侵袭、迁移的影响及分子机制。方法利用癌症基因组图谱(TCGA)及基因表达综合数据库(GEO)联合分析胃癌中与细胞侵袭相关差异表达基因,京都基因和基因组百科全书(KEGG)分析差异基因所富集的通路,根据差异倍数及功能确定候选基因LAMC2,GEPIA(Gene Expression Profiling Interactive Analysis)在线数据库分析胃癌中LAMC2的表达,采用蛋白印迹法(Western blot)检测人胃癌细胞株中LAMC2的表达。构建LAMC2敲减胃癌细胞模型,Western blot检测胃癌细胞中LAMC2的表达,将敲减模型构建成功的胃癌细胞(AGS、NCI-N87)分成空白对照组(Control)、阴性对照组(si-NC)及LAMC2敲减组(si-LAMC2),Transwell法检测各组胃癌细胞的侵袭及迁移能力,Western blot检测各组胃癌细胞中EMT标志物(E-cadherin、N-cadherin、Vimentin)表达及ERK通路激活情况。结果LACM2在胃癌中的表达显著上调,KEGG分析其主要富集于细胞外基质受体通路(ECM-receptor interaction)。与GES-1细胞比较,人胃癌细胞(KATOⅢ、AGS、NCI-N87、SNU-1)中LACM2的表达均显著升高(均P=0.000),其中AGS表达最低,NCI-N87表达最高。在AGS胃癌细胞中,与si-NC组比较,si-LAMC2显著抑制胃癌细胞侵袭(67.000±4.003 vs 19.675±6.032,P=0.000)及迁移[(64.723±3.062)vs(18.655±4.040),P=0.000],N-cadherin[(0.535±0.041)vs(0.209±0.028),P=0.000]及Vimentin[(0.840±0.032)vs(0.286±0.056),P=0.000]的表达被显著抑制,而E-cadherin[(0.333±0.025)vs(0.570±0.019),P=0.000]的表达显著升高,ERK磷酸化水平(0.638±0.031 vs 0.243±0.027,P=0.000)被显著下调。类似地,抑制LAMC2后,NCI-N87细胞的侵袭、迁移能力倍显著抑制,ERK通路被显著抑制。结论LAMC2在胃癌中的表达上调,其可能通过激活ERK通路促进胃癌细胞侵袭及迁移。Objective To study the effects of laminin subunit gamma 2(LAMC2)on the invasion and migration of gastric cancer cells and its molecular mechanism.Methods Cancer Genome Atlas(TCGA)and gene expression database(GEO)were used to analyze differentially expressed genes related to cell invasion in gastric cancer.The Kyoto Encyclopedia of Genes and Genomes(KEGG)was applied to analyze the pathways in which different Genes are enriched.Candidate gene LAMC2 was identified according to differential expression and function.Gene Expression Profiling Interactive Analysis(GEPIA)database was used to analyze the expression of LAMC2 in gastric cancer cells,and Western blot was used to detect the expression of LAMC2 in human gastric cancer cell lines.LAMC2 knockout gastric cancer cell model was constructed,and the expression of LAMC2 in gastric cancer cells was detected by Western blot.The gastric cancer cells successfully constructed by knockout model(AGS and NCI-N87)were divided into blank Control group(Control),negative Control group(si-NC),and LAMC2 knockout group(si-LAMC2).The invasion and migration ability of gastric cancer cells in each group were detected by Transwell,and the expression of EMT markers(E-cadherin,N-cadherin,Vimentin)and the activation of ERK pathway in gastric cancer cells in each group were detected by Western blot.Results The expression of LACM2 was significantly up-regulated in gastric cancer,and it was mainly enriched in ECM-receptor interaction according to KEGG analysis.Compared with GES-1 cells,the expression of LACM2 in human gastric cancer cells(KATOⅢ,AGS,NCI-N87,and SNU-1)was significantly increased(all P=0.000),in which AGS was the lowest and NCI-N87 was the highest.Compared with si-NC group,si-LAMc2 significantly inhibited the migration[(67.000±4.003)vs(19.675±6.032),P=0.000]and invasion[(64.723±3.062)vs(18.655±4.040),P=0.000]of gastric cancer cells,inhibited the expression of N-cadherin[(0.535±0.041)vs(0.209±0.028),P=0.000]and Vimentin[(0.840±0.032)vs(0.286±0.056),P=0.000],increased the

关 键 词：胃癌 层黏连蛋白γ_(2) 上皮-间质转化 侵袭及转移 细胞外调节蛋白激酶 

分 类 号：R735.2[医药卫生—肿瘤]