摘要
目的 探讨lncRNA HOXA11-AS调节miR-152-3p/高迁移率族蛋白A2(HMGA2)轴对非小细胞肺癌(NSCLC)恶性生物学行为的影响。方法 qRT-PCR检测NSCLC组织和细胞中HOXA11-AS、miR-152-3p和HMGA2 mRNA的表达水平。NSCLC细胞16HBE分为对照组、si-NC组、si-HOXA11-AS组、si-HOXA11-AS+inhibitor NC组、si-HOXA11-AS+miR-152-3p inhibitor组、siHOXA11-AS+oe-NC组、si-HOXA11-AS+oe-HMGA2组。qRT-PCR检测各组细胞中HOXA11-AS、miR-152-3p和HMGA2 mRNA表达水平,克隆形成实验检测细胞克隆能力,细胞划痕实验检测细胞迁移,Transwell实验检测细胞侵袭,流式细胞术检测细胞凋亡率,Western blot检测细胞中E-cadherin、N-cadherin、vimentin、Bax、PCNA、MMP-2、HMGA2蛋白表达水平,双荧光素酶报告实验验证miR-152-3p与HOXA11-AS和HMGA2的靶向关系。结果 与癌旁组织相比,NSCLC组织中HOXA11-AS和HMGA2mRNA的表达升高(P<0.05),miR-152-3p表达降低(P<0.05)。与正常肺上皮BEAS-2B细胞相比,NSCLC细胞中HOXA11-AS和HMGA2 mRNA的表达升高,miR-152-3p表达降低(P<0.05)。与对照组和si-NC组相比,si-HOXA11-AS组16HBE细胞中HOXA11-AS和HMGA2 mRNA水平、细胞克隆形成率、划痕愈合率和侵袭个数、N-cadherin、vimentin、PCNA、MMP-2、HMGA2蛋白表达水平降低(P<0.05),miR-152-3p表达、细胞凋亡率、E-cadherin和Bax蛋白表达水平升高(P<0.05)。下调miR-152-3p表达或上调HMGA2表达均可减弱沉默HOXA11-AS表达对16HBE细胞恶性生物学行为的抑制作用(P<0.05)。结论 沉默HOXA11-AS表达可通过升高miR-152-3p表达,降低HMGA2表达来抑制16HBE细胞增殖和迁移,并促进细胞凋亡。
Objective To investigate the effect of lncRNA HOXA11⁃AS on the malignant biological behaviors of non⁃small cell lung cancer(NSCLC)by regulating the miR⁃152⁃3p/high mobility group protein A2(HMGA2)axis.Methods qRT⁃PCR was applied to detect the expression levels of HOXA11⁃AS,miR⁃152⁃3p,and HMGA2 mRNA in NSCLC tissue and cells.16HBE cells were divided into control group,si⁃NC group,si⁃HOXA11⁃AS group,si⁃HOXA11⁃AS+inhibitor NC group group,si⁃HOXA11⁃AS+miR⁃152⁃3p in⁃hibitor group,si⁃HOXA11⁃AS+oe⁃NC group,and si⁃HOXA11⁃AS+oe⁃HMGA2 group.qRT⁃PCR was applied to detect expression le⁃vels of HOXA11⁃AS,miR⁃152⁃3p,and HMGA2 mRNA in cells.Clone formation experiment was applied to test the cell cloning abi⁃lity.Cell scratch assay was applied to detect the cell migration.Transwell was applied to detect the cell invasion.Flow cytometry was applied to detect the rate of cell apoptosis.Western blot was applied to detect the protein expression levels of E⁃cadherin,N⁃cadherin,vimentin,Bax,PCNA,MMP⁃2,and HMGA2 in cells.Dual luciferase reporter assay was applied to verify the targeting relationship of miR⁃152⁃3p with HOXA11⁃AS and HMGA2.Results Compared with adjacent tissues,the expressions of HOXA11⁃AS and HMGA2 mRNA in NSCLC tissues were increased(P<0.05),while the expression of miR⁃152⁃3p was decreased(P<0.05).Compared with normal lung epithelial BEAS⁃2B cells,the expressions of HOXA11⁃AS and HMGA2 mRNA were increased while the expression of miR⁃152⁃3p was decreased in NSCLC cells(P<0.05).Compared with control group and si⁃NC group,the expression levels of HOXA11⁃AS and HMGA2 mRNA,the cell clone formation rate,the scratch healing rate,the number of invasive cells,and the pro⁃tein expression levels of N⁃cadherin,vimentin,PCNA,MMP⁃2,and HMGA2 in 16HBE cells were decreased in si⁃HOXA11⁃AS group(P<0.05),while the expression of miR⁃152⁃3p,the cell apoptosis rate,and the expression levels of E⁃cadherin and Bax proteins were increased
作者
张进召
李亚明
潘双
刘松
ZHANG Jinzhao;LI Yaming;PAN Shuang;LIU Song(Department of Intensive Care Medicine,First Affiliated Hospital of Xi'an Medical University,Xi'an 710000,China;Department of Respiratory and Critical Care Medicine,First Affiliated Hospital of Xi'an Medical University)
出处
《山西医科大学学报》
CAS
2024年第11期1388-1397,共10页
Journal of Shanxi Medical University
基金
陕西省重点研发计划项目(2022SF-554,2021SF-039)。
