lncRNA RP11-297P16.4促进肺腺癌细胞侵袭及迁移的机制探讨  

Mechanism of lncRNARP11-297P16.4 promoting invasion and migration of lung adenocarcinoma cells

作　　者：徐白雪 王维 庞敏[2] 樊莎莎 王智欣 张力中 安彩兴 张沛霖 程保龙 刘芳芳 王海龙 于保锋 XU Baixue;WANG Wei;PANG Min;FAN Shasha;WANG Zhixin;ZHANG Lizhong;AN Caixing;ZHANG Peilin;CHENG Baolong;LIU Fangfang;WANG Hailong;YU Baofeng(Department of Biochemistry and Molecular Biology,School of Basic Medicine,Basic Medical Sciences Center,Shanxi Medical University,Shanxi Jinzhong 030600,China;Department of Pulmonary and Critical Care Medcine,the First Hospital,Shanxi Medical University,Shanxi Province Key Laboratory of Respiratory Disease,Shanxi Taiyuan 030001,China)

机构地区：[1]山西医科大学基础医学院生物化学与分子生物学教研室,基础医学研究中心,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

出　　处：《现代肿瘤医学》2023年第3期395-401,共7页Journal of Modern Oncology

基　　金：全国大学生创新训练项目(编号:202010114013)。

摘　　要：目的:探究长链非编码RNA(long non-coding RNA,lncRNA)RP11-297P16.4对人肺腺癌细胞H1299和A549侵袭及迁移的影响。方法:利用qRT-PCR检测在肺腺癌细胞H1299、A549及人正常支气管上皮细胞HBEC中RP11-297P16.4的相对表达水平;设计合成针对RP11-297P16.4的特异性shRNA,慢病毒感染建立稳定敲低lncRNA RP11-297P16.4的H1299、A549细胞;将其分为对照/H1299组、实验/H1299组、对照/A549组、实验/A549组,对照组感染shRNA-NC,实验组感染shRNA-RP11-297P16.4;qRT-PCR检测shRNA敲低效率;采用细胞划痕实验和Transwell实验检测RP11-297P16.4对肺腺癌细胞侵袭、迁移能力的影响;采用qRT-PCR和蛋白质印迹法检测基质金属蛋白酶(matrix metalloproteinase,MMP)-2和-9的mRNA和蛋白质表达水平。结果:与对照组相比,RP11-297P16.4在肺腺癌细胞中高表达,差异有统计学意义(P<0.001,P<0.01);qRT-PCR分析确定,shRNA敲低后RP11-297P16.4的mRNA表达水平显著降低(P<0.001,P<0.001);划痕实验表明24 h、48 h实验组细胞伤口愈合率均低于对照组,差异有统计学意义(P<0.05,P<0.01;P<0.001,P<0.001);Transwell小室检测结果显示,实验组细胞侵袭数量显著少于对照组,差异有统计学意义(P<0.001,P<0.001);qRT-PCR和蛋白印迹法结果显示,实验组细胞中MMP-2和MMP-9 mRNA和蛋白质表达量均低于对照组,差异具有统计学意义(均P<0.05)。结论:lncRNA RP11-297P16.4通过促进MMP-2和MMP-9 mRNA和蛋白表达来增强H1299、A549细胞的侵袭和迁移能力。Objective:To investigate the effect of long non-coding RNA(lncRNA)RP11-297P16.4 on the invasion and migration of human lung adenocarcinoma cells of H1299 and A549.Methods:The relative expression levels of RP11-297P16.4 in H1299,A549 and human normal bronchial epithelial cells HBEC were detected by real-time fluorescence quantitative PCR.Special shRNA for RP11-297P16.4 was designed and synthesized.H1299,A549 cells with stable knockdown of lncRNA RP11-297P16.4 were established by lentivirus infection,then they were divided into control/H1299 group,experimental/H1299 group,control/A549 group and experimental/A549 group.The control group was infected with shRNA-NC,and the experimental group was infected with shRNA-RP11-297P16.4.qRT-PCR was used to detect the shRNA inhibition efficiency.Cell scratch and Transwell assays were used to establish the effect of RP11-297P16.4 on the invasion and migration of lung adenocarcinoma cells.qRT-PCR andWestern blot were performed to assay the mRNA and protein expression levels of matrix metalloproteinase(MMP)-2 and-9.Results:Compared with the control group,RP11-297P16.4 was highly expressed in lung adenocarcinoma cells,and the difference was statistically significant(P<0.001,P<0.01).qRT-PCR analysis determined that the mRNA expression level of RP11-297P16.4 was significantly reduced after shRNA knockdown(P<0.001,P<0.001).The scratch assay demonstrated that the wound healing rate of cells in the experiment group at 24 h and 48 h was lower than that in the control group,and the difference was statistically significant(P<0.05,P<0.01,P<0.001,P<0.001).Transwell experiment showed that the number of cellular invasion and migration in the experiment group was significantly less than that in the control group,and the difference was statistically significant(P<0.001,P<0.001).The results of qRT-PCR andWestern blot revealed that the mRNA and protein expression levels of MMP-2 and MMP-9 in the experiment group were lower than those in the control group,and the difference was statistically signi

关 键 词：肺腺癌 lncRNA RP11-297P16.4 侵袭 迁移 基质金属蛋白酶-2 基质金属蛋白酶-9 

分 类 号：R734.2[医药卫生—肿瘤]