LncRNAHOTAIRM1/HOXA1促进胶质瘤增殖迁移和侵袭

Role of LncRNA HOTAIRM1/HOXA1 in proliferation,migration and invasion of glioma

目的探究长链非编码RNA HOTAIRM1(LncRNA HOTAIRM1)靶向调控HOXA1对胶质瘤细胞增殖、迁移和侵袭的影响。方法收集2020年10月至2021年12月于温州市中心医院接受手术治疗的60例胶质瘤患者的肿瘤组织及癌旁正常脑组织标本,采用实时荧光定量聚合酶链式反应(qRT-PCR)测定并比较正常脑组织和胶质瘤组织中HOTAIRM1 mRNA和HOXA1 mRNA表达;构建干扰LncRNA HOTAIRM1的慢病毒载体(sh-HOTAIRM1)及其阴性对照载体(sh-NC)、HOXA1过表达载体(pcDNA3.1-HOXA1)及其阴性对照载体(pcDNA3.1-NC)。培养人胶质瘤细胞LN229并进行转染干预,分为对照组(无转染)、sh-HOTAIRM1组(转染sh-HOTAIRM1)、sh-NC组(转染sh-NC)、sh-HOTAIRM1+HOXA1组(转染sh-HOTAIRM1和pcDNA3.1-HOXA1)、sh-HOTAIRM1+NC组(转染sh-HOTAIRM1和pcDNA3.1-NC)。采用MTT法检测转染后各组细胞增殖能力,采用流式细胞仪检测各组细胞凋亡水平,采用Transwell实验测定各组细胞侵袭和迁移能力。结果胶质瘤组织中HOTAIRM1 mRNA和HOXA1 mRNA的表达水平高于正常脑组织(均P<0.05);LN229细胞转染48h后,sh-HOTAIRM1组和sh-HOTAIRM1+NC组细胞中HOTAIRM1 mRNA和HOXA1 mRNA表达水平均低于对照组和sh-NC组(均P<0.05),细胞凋亡率高于对照组和sh-NC组(均P<0.05),迁移细胞数与侵袭细胞数均少于对照组和sh-NC组(均P<0.05);sh-HOTAIRM1+HOXA1组细胞中HOXA1 mRNA表达水平高于sh-HOTAIRM1组和sh-HOTAIRM1+NC(均P<0.05),凋亡率低于sh-HOTAIRM1组和sh-HOTAIRM1+NC组(均P<0.05),细胞迁移细胞数与侵袭细胞数均多于sh-HOTAIRM1组和sh-HOTAIRM1+NC组(均P<0.05)。转染后24~72h,各组细胞增殖活性均逐渐升高(均P<0.05);在不同时间点,sh-HOTAIRM1组和sh-HOTAIRM1+NC组细胞增殖活性均弱于对照组和sh-NC组(均P<0.05),sh-HOTAIRM1+HOXA1组细胞增殖活性强于sh-HOTAIRM1组和sh-HOTAIRM1+NC组(均P<0.05)。结论LncRNAHOTAIRM1和HOXA1在胶质瘤组织中呈高表达状态,HOTAIRM1或可通过靶向调控HOXA1促进胶质瘤细胞增殖,抑制其凋亡,并增强胶质瘤细胞迁移和侵袭能力。

Objective To investigate the effect of long non-coding RNA HOTAIRMI(LncRNA HOTAIRMI1)targeted regulation of HOXAI on proliferation,migration and invasion of glioma cells.Methods Tumor tissues and matched normal brain tissues were collected from 60 patients with glioma who underwent operation in Wenzhou Central Hospital from October 2020 to December 2021.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to determine and compare the expression ofHOTAIRM1 mRNA and HOXA1 mRNA in human normal brain tissue and glioma tissue.The lentiviral vector(sh-HOTAIRMI)and its negative control vector(sh-NC),HOXA1 overexpression vector(pcDNA3.1-HOXAI)and its negative control vector(pcDNA3.1-NC)for interfering with LncRNA HOTAIRM1 were constructed.Human glioma cell line LN229 was cultured,and given transfection intervention.Then,the cells were divided into control group(no transfection),sh-HOTAIRM1 group(transfected by sh-HOTAIRM1),sh-NC group(transfected by sh-NC),sh-HOTAIRM1+HOXA1 group(transfected by sh-HOTAIRM1 and pcDNA3.1-HOXA1),and sh-HOTAIRM1+NC group(transfected by sh-HOTAIRM1 and pcDNA3.1-NC).MTT assay was used to detect cell proliferation in each group after transfection,and flow cytometry was used to detect apoptosis in each group.Transwell assay was used to determine the invasion and migration ability of cells in each group.Results HOTAIRM1 mRNA and HOXAI mRNA expression levels were higher in glioma tissues than normal brain tissues(all P<0.05).After 48h oftransfection,HOTAIRM1 mRNA and HOXA1 mRNA expression levels were significantly lower in the sh-HOTAIRMI group and the sh-HOTAIRM1+NC group than the control group and the sh-NC group(all P<0.05).HOXA1 mRNA expression level was significantly higher in the sh-HOTAIRM1+HOXA1 group than the sh-HOTAIRMI group and the sh-HOTAIRM1+NC group(all P<0.05).From 24h to 72h after transfection,the proliferation activity of cells in all groups gradually increased(all P<0.05).At different time points,the proliferation activity of the sh-HOTAIRM1 group and the sh-HOTAIRM1+NC group was significantly lower when compared with that of the control group and the sh-NC group(all P<0.05).The proliferation activity of the sh-HOTAIRMI+HOXA1 group was significantly greater when compared with that of the sh-HOTAIRM1 group and the sh-HOTAIRMI+NC group(all P<0.05).At different time points,the cell proliferation activity of sh-HOTAIRMI group and sh-HOTAIRMI+NC group was weaker than that of control group and sh-NC group(all P<0.05).The cell proliferation activity of sh-HOTAIRM1+HOXA1 group was stronger than that of sh-HOTAIRM1 group and sh-HOTAIRMI+NC group(all P<0.05).Conclusions LncRNA HOTAIRM1 and HOXA1 are highly expressed in glioma tissues.HOTAIRM1 maybecan promote the proliferation of glioma cells,inhibit their apoptosis,and enhance the migration and invasion of glioma cells by targeting HOXA1.