lncRNA HCG11-201对肾癌细胞增殖和侵袭的影响

Effect of lncRNA HCG11-201 on the proliferation and invasion of renal cancer cells

目的探讨长链非编码RNA(lncRNA)HCG11-201在肾癌组织中的表达及影响癌细胞增殖和侵袭的分子机制。方法生物信息学方法预测HCG11-201表达与肾癌患者预后的相关性。实时荧光定量PCR(qPCR)检测肾癌组织和癌旁正常组织、肾癌细胞系和正常肾小管上皮细胞中HCG11-201的表达,选择表达最少的肾癌细胞系转染,分别将HCG11-201质粒(实验组)或阴性对照质粒(对照组)转至肾癌细胞。qPCR检测HCG11-201质粒转染效率。MTT法检测转染后细胞增殖活性,Transwell小室实验检测转染后细胞侵袭能力。生物信息学方法预测HCG11-201可吸附结合的微小RNA(miRNA)及miRNA下游基因。qPCR和Western blot检测miRNA和下游基因表达。结果GEPIA数据库显示,HCG11-201相对表达水平越高,患者总生存期越长(P<0.01)。肾癌组织HCG11-201相对表达水平显著低于癌旁正常组织(P<0.01)。肾癌细胞HCG11-201相对表达水平显著低于正常肾小管上皮细胞(P<0.01),其中Caki-1细胞相对表达水平最低(P<0.01)。与对照组比较,实验组Caki-1细胞HCG11-201相对表达水平显著升高(P<0.01),细胞增殖活性显著降低(P<0.05),细胞的侵袭能力显著降低(P<0.05)。HCG11-201可互补结合miR-522,miR-522可互补结合线粒体融合蛋白2(mitofusion-2)。与对照组比较,实验组Caki-1细胞miR-522的相对表达水平显著降低(P<0.01),mitofusion-2的mRNA相对表达水平和蛋白水平显著升高(P<0.01)。结论lncRNA HCG11-201在肾癌组织和细胞系低表达,与肾癌患者预后相关。过表达HCG11-201可明显抑制肾癌细胞增殖和侵袭,其分子机制可能为HCG11-201吸附miR-522进而上调mitofusion-2基因的表达。

Objective To investigate the expression of long-chain non-coding RNA(lncRNA)HCG11-201 in renal cancer tissues and the molecular mechanisms that affect the proliferation and invasion of renal cancer cells.Methods Bioinformatics predicts the correlation between HCG11-201 expression level and prognosis of patients with renal cancer real-time fluorescent quantitative polymerase chain reaction(qPCR)was used to detect the expression of HCG11-201 in renal cancer tissues and adjacent normal tissues,renal cancer cell lines,and normal renal tubular epithelial cells.The lowest expressing renal cancer cell line was selected for transfection,and tuhe HCG11-201 plasmid(experimental group)or negative control plasmid(control group)were transferred into renal cancer cells,respectively.qPCR was used to detect the transfection efficiency of HCG11-201 plasmid.The MTT method was used to detect the proliferation activity of the cells after transfection,and the Transwell assay was used to detect the invasion ability of the cells after transfection.Bioinformatics method predicted that HCG11-201 could adsorb binding microRNA(miRNA)and miRNA downstream gene.qPCR and Western blot were used to detect the expression of miRNA and downstream gene.Results GEPIA database showed that,the higher the relative expression level of HCG11-201,the longer the overall survival of patients with renal cancer(P<0.01).The relative expression level of HCG11-201 in renal cancer tissues was significantly lower than that in adjacent normal tissues(P<0.01).The relative expression level of HCG11-201 in renal cancer cells was significantly lower than that in normal renal tubular epithelial cells(P<0.01),and the expression level of Caki-1 cells was the lowest(P<0.01).Compared with the control group,the relative expression level of HCG11-201 in Caki-1 cells in the experimental group was significantly increased(P<0.01),the cell proliferative activity was significantly reduced(P<0.05),and the cell invasion capacity was significantly reduced(P<0.05).HCG11-201 could complement miR-522,and miR-522 could complement mitofusion-2.Compared with the control group,the relative expression level of miR-522 in Caki-1 cells in the experimental group was significantly reduced(P<0.01),and the relative expression level of mitofusion-2 in mRNA and protein levels was significantly increased(P<0.01).Conclusion HCG11-201 is low-expressed in renal cancer tissues and cell lines,and is related to the prognosis of renal cancer patients.Overexpression of HCG11-201 can inhibit the proliferation and invasion of renal cancer cells,and its molecular mechanism may be that HCG11-201 specifically adsorbs miR-522 and then up-regulates the expression of mitofusion-2 gene.