lncRNA TMPO-AS1靶向miR-1224-5p调控肝癌细胞的增殖和凋亡

lncRNA TMPO-AS1 targeting miR-1224-5p to regulate the proliferation and apoptosis of liver cancer cells

目的研究长链非编码RNA(lncRNA)TMPO antisense RNA 1(TMPO-AS1)与微小RNA(miRNA)-1224-5p的靶向关系及对肝癌细胞的增殖和凋亡的影响。方法实时荧光定量PCR(qRT-PCR)测定lncRNA TMPO-AS1和miR-1224-5p在肝癌组织中的表达。生物信息学预测和双荧光素酶报告基因检测分析lncRNA TMPO-AS1对miR-1224-5p的靶向调控。在肝癌HCCLM3细胞中转染si-TMPO-AS1或miR-1224-3p或共转染si-TMPO-AS1和anti-miR-1224-5p。qRT-PCR测定lncRNA TMPO-AS1和miR-1224-5p表达,噻唑蓝(MTT)比色法、流式细胞术和免疫印迹实验(western blot)分别测定肝癌HCCLM3细胞增殖、凋亡与细胞周期蛋白D1(CyclinD1)、p21、B细胞淋巴瘤/白血病-2(Bcl-2)和抗Bcl-2相关X蛋白(Bax)蛋白的表达。结果肝癌组织中lncRNA TMPO-AS1表达上调,miR-1224-5p表达下调,差异有统计学意义(P<0.05)。lncRNA TMPO-AS1靶向调控miR-1224-5p的表达。敲低lncRNA TMPO-AS1表达明显降低细胞增殖、CyclinD1和Bcl-2蛋白表达,显著提高细胞凋亡、p21和Bax蛋白表达,差异有统计学意义(P<0.05),与miR-1224-5p过表达的结果一样。抑制miR-1224-5p表达后,敲低lncRNA TMPO-AS1表达对肝癌HCCLM3细胞增殖的抑制作用和对细胞凋亡的促进作用被逆转。结论lncRNA TMPO-AS1在肝癌组织表达上调,敲低其表达通过靶向miR-1224-5p抑制肝癌细胞的增殖,并诱导细胞凋亡。

Objective To investigate the targeting correlation between long-chain non-coding RNA(lncRNA)TMPO antisense RNA 1(TMPO-AS1)and microRNA(miRNA)-1224-5p,and its effects on the proliferation and apoptosis of liver cancer cells.Methods Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression levels of lncRNA TMPO-AS1 and miR-1224-5p in liver cancer tissue.Bioinformatics prediction with dual luciferase reporter gene detection was used to analyze the targeted regulation of miR-1224-5p by lncRNA TMPO-AS1.The HCCLM3 cells were transfected by si-TMPO-AS1 or miR-1224-3p,or co-transfect si-TMPO-AS1 and anti-miR-1224-5p.Moreover qRT-PCR was performed to determine the expression levels of lncRNA TMPO-AS1 and miR-1224-5p.MTT colorimetry,flow cytometry and Western Blot were used to detect the proliferation,apoptosis and the expressions CyclinD1,p21,Bcl-2 Bax protein.Results The expression levels of lncRNA TMPO-AS1 were up-regulated,however,the expression levels of miR-1224-5p were down-regulated in liver cancer tissues(P<0.05).lncRNA TMPO-AS1 targetedly regulated the expression of miR-1224-5p.And knocking down lncRNA TMPO-AS1 expression could significantly decrease cell proliferation,and the expressions of CyclinD1 and Bcl-2 protein,could significantly increase the expression levels of p21 and Bax protein(P<0.05),so could by miR-1224-5p overexpression.After inhibiting the expression of miR-1224-5p,knocking down the expression of lncRNA TMPO-AS1 could reverse the inhibitory effects on the proliferation of HCCLM3 cells and the promotive effects on cell apoptosis.Conclusion The expression levels of lncRNA TMPO-AS1 are up-regulated in liver cancer tissue,and knocking down its expression can inhibit the proliferation of liver cancer cells and induce apoptosis by targeting miR-1224-5p.