MTUS2-AS2在膀胱癌组织中的表达及对膀胱癌细胞增殖和侵袭的影响

Expression of IncRNA MTUS2-AS2 in bladder cancer tissues and its effect on proliferation and invasion of bladder cancer

目的探讨长链非编码RNA(lncRNA)MTUS2-AS2在膀胱癌组织中的表达及其对膀胱癌细胞增殖和侵袭的影响。方法荧光实时定量PCR(qRT-PCR)检测63例膀胱癌组织和癌旁组织、膀胱癌细胞系(BIU-87、J82、T24、5637)和正常膀胱上皮细胞系SV-HUC-1中的MTUS2-AS2表达。以MTUS2-AS2表达最低的J82细胞进行研究,分为对照组(转染载有无意义序列的对照质粒)和实验组(转染载有MTUS2-AS2表达序列的质粒)。qRT-PCR检测转染效率。MTS法和Transwell侵袭实验检测J82细胞的增殖和侵袭能力。qRT-PCR检测J82细胞中趋化素样因子超家族成员5(CMTM5)mRNA的表达,Western blot检测两组细胞中CMTM5、PI3K-AKT信号通路蛋白的表达。结果膀胱癌组织MTUS2-AS2的表达低于癌旁组织(P<0.01),膀胱癌细胞系MTUS2-AS2的表达明显低于正常膀胱上皮细胞(P<0.05),J82细胞中MTUS2-AS2的表达最少(P<0.01)。与对照组相比,实验组J82细胞中MTUS2-AS2的表达明显增加(P<0.01)。与对照组相比,实验组J82细胞增殖活力明显降低(P<0.05),细胞侵袭能力明显降低(P<0.01)。与对照组相比,实验组J82细胞中CMTM5 mRNA和蛋白表达增加(P<0.01),PI3K-AKT信号通路蛋白表达降低(P<0.05)。结论 MTUS2-AS2在膀胱癌组织和细胞系中呈低表达,MTUS2-AS2可通过上调CMTM5基因的表达,抑制J82细胞的增殖和侵袭能力。

Objective To obsene the expression of long non-coding RNA(IncRNA)MTUS2-AS2 in bladder cancer tissues and its effect on proliferation and invasion of bladder cancer cells.Methods Fluorescence real-time quantitative PCR(qRT-PCR)was used to detect MTUS2-AS2 expression in 63 bladder cancer tissues and paracancerous tissues,bladder cancer cell lines(BIU-87,J82,T24,5637)and normal bladder epithelial cell line SV-HUC-1.The J82 cells with the lowest expression of MTUS2-AS2 were used for the study,and were divided into control group(transfected with control plasmid carrying non-significant sequences)and experimental group(transfected with plasmid carrying the expression sequence of MTUS2-AS2).Pransfection efficiency was detected by qKT-PCR.The proliferation and invasion ability of J82 cells were detected by MTS assay and Transwell invasion assay.The expression of CMTM5 mRNA in J82 cells was detected by qRT-PCR.The expression of CMTM5 and PI3K-AKT signaling pathway proteins were detected by Western blot.Results The expression of MTUS2-AS2 in bladder cancer was significantly lower than that in adjacent tissues(P<0.01).The expression of MTUS2-AS2 in bladder cancer cells was significantly lower than that in bladder tubular epithelial cells(P<0.05).The expression of MTLIS2-AS2 was the lowest in J82 cells(P<0.01).Compared with the control group,the expression of MTUS2-AS2 in J82 cells was significantly increased after transfection with MTUS2-AS2 plasmid(P<0.01).Compared with the control group,the proliferation of J82 cells in the experimental group was significantly decreased(P<0.05),and the cell invasion ability was significantly decreased(P<0.01).Compared with the control group,the expression of CMTM5 mRNA in the experimental group was significantly increased(P<0.01).The expression levels of CMTM5 were significantly up-regulated(P<0.01),and PI3K-AKT signaling pathway proteins were significantly down-regulated(P<0.05).Conclusions The expression of MTUS2-AS2 is decreased in bladder cancer tissues and cell lines.MTUS2-AS2 can inhibit the proliferation and invasion of J82 cells by upregulating the expression of CMTM5 gene.