lncRNA AL022344.4促进肺腺癌细胞侵袭及迁移的初步研究  

Preliminary study on the role of the lncRNA AL022344.4 in promoting invasion and migration of lung adenocarcinoma cells

作　　者：倪林峰 王维 庞敏[2] 刘超锋[2] 王海龙[1] 孙焱 NI Linfeng;WANG Wei;PANG Min;LIU Chaofeng;WANG Hailong;SUN Yan(Institute of Cancer Biology,Basic Medical Sciences Center,School of Basic Medicine,Shanxi Medical University,Shanxi Jinzhong 030600,China;Shanxi Province Key Laboratory of Respiratory Disease Control,Department of Respiratory and Critical Care Medicine,the First Hospital,Shanxi Medical University,Shanxi Taiyuan 030001,China;Department of Management,Shanxi Medical University,Shanxi Jinzhong 030600,China.)

机构地区：[1]山西医科大学基础医学院,基础医学研究中心,肿瘤生物学研究所,山西晋中030600 [2]山西医科大学第一医院呼吸与危重症医学科,呼吸疾病防治山西省重点实验室,山西太原030001 [3]山西医科大学管理学院,山西晋中030600

出　　处：《现代肿瘤医学》2024年第14期2523-2529,共7页Journal of Modern Oncology

基　　金：山西省自然科学基金项目(编号:202203021221184)。

摘　　要：目的:研究长链非编码RNA(long non-coding RNA,lncRNA)AL022344.4对人肺腺癌细胞H157和A549侵袭及迁移的影响。方法:选取TCGA_LUAD和GTEx_LUNG两个公共数据库中公开数据集对lncRNA AL022344.4表达数据进行生物信息学分析;qRT-PCR检测人正常支气管上皮细胞HBEC及肺腺癌细胞(H157、A549、H1299、H1975、CALU-3)中AL022344.4的相对表达量。慢病毒感染及质粒转染构建AL022344.4敲低H157细胞及过表达AL022344.4的A549细胞,qRT-PCR检测AL022344.4在两种细胞系中的表达;采用细胞划痕实验和Transwell实验检测AL022344.4敲低及过表达对肺腺癌细胞迁移、侵袭能力的影响;采用蛋白印迹法检测E-Cadherin、N-Cadherin、Vimentin、α-SMA、MMP9和ZEB1蛋白质表达水平。结果:生物信息学分析发现lncRNA AL022344.4在肺腺癌中较匹配的癌旁组织表达更高,qRT-PCR表明其在肺腺癌细胞中的表达亦高于正常支气管上皮细胞,差异有统计学意义(P<0.01);qRT-PCR表明AL022344.4敲低H157细胞系及过表达A549细胞系构建成功;细胞划痕和Transwell实验显示过表达AL022344.4促进A549细胞的迁移和侵袭,敲低AL022344.4抑制H157细胞迁移和侵袭;蛋白印迹法结果显示,过表达AL022344.4促进N-Cadherin、α-SMA、Vimentin、MMP9和ZEB1表达,降低E-Cadherin表达;敲低AL022344.4促进E-Cadherin表达,降低N-Cadherin、Vimentin、α-SMA、MMP9和ZEB1表达。结论:lncRNA AL022344.4通过促进ZEB1表达调控E-Cadherin、N-Cadherin、Vimentin和α-SMA,协同MMP9共同促进肺腺癌细胞的侵袭和迁移。Objective:To investigate the impact of the long non-coding RNA(lncRNA)AL022344.4 on the invasion and migration of human lung adenocarcinoma cells H157 and A549.Methods:Two publicly available datasets from TCGA_LUAD and GTEx_LUNG were selected for bioinformatics analysis of lncRNA AL022344.4 expression.qRT-PCR was used to detect the relative expression of AL022344.4 in human normal bronchial epithelial cells(HBEC)and several lung adenocarcinoma cells(H157,A549,H1299,H1975,CALU-3).Lentiviral infection and plasmid transfection were employed to establish AL022344.4 knockdown in H157 cells and AL022344.4 overexpression in A549 cells.qRT-PCR was performed to confirm AL022344.4 expression in both cell lines.Cell scratch and Transwell experiments were conducted to assess the impact of AL022344.4 knockdown or overexpression on the migration and invasion of H157 and A549 cells.Western blot analysis was used to measure the protein levels of E-Cadherin,N-Cadherin,Vimentin,α-SMA,MMP9,and ZEB1.Results:Bioinformatics analysis revealed higher expression of lncRNA AL022344.4 in lung adenocarcinoma tissues compared to matched adjacent tissues.qRT-PCR confirmed its elevated expression in lung adenocarcinoma cells compared to HBEC cells,with statistical significance(P<0.01).qRT-PCR further demonstrated that the AL022344.4 knockdown H157 cells or AL022344.4 overexpression A549 cells were successfully constructed.Cell scratch and Transwell experiments showed that AL022344.4 overexpression promoted migration and invasion of A549 cells,while AL022344.4 knockdown inhibited migration and invasion of H157 cells.Western blot indicated that AL022344.4 overexpression increased the protein levels of N-Cadherin,α-SMA,Vimentin,MMP9,and ZEB1,while reduced E-Cadherin level.Conversely,AL022344.4 knockdown enhanced E-Cadherin level,but decreased the protein levels of N-Cadherin,Vimentin,α-SMA,MMP9,and ZEB1.Conclusion:lncRNA AL022344.4 promotes the expression of ZEB1 to regulate E-Cadherin,N-Cadherin,Vimentin,α-SMA,and collaboratively with MMP9,e

关 键 词：肺腺癌 lncRNA AL022344.4 侵袭 迁移 锌指结构E-box-结合同源框1 上皮-间质转化 MMP9 

分 类 号：R734.2[医药卫生—肿瘤]