lncRNA ZBED3-AS1通过吸附miR-512-5p调控DACH1抑制前列腺癌细胞的增殖和迁移

lncRNA ZBED3-AS1 suppresses proliferation and migration of prostate cancer cells by adsorption of miR-512-5p and regulating DACH1

目的分析长链非编码RNA(long-chain non-coding RNA,lncRNA)ZBED3-AS1通过调节miR-512-5p在前列腺癌发生、发展中的作用。方法qRT-PCR技术检测67例前列腺癌组织及相应癌旁正常组织中ZBED3-AS1的表达并分析前列腺癌细胞株及永生化前列腺上皮细胞中ZBED3-AS1的表达。将表达最低的细胞根据随机数字法分为阴性对照组(转染阴性对照质粒)和ZBED3-AS1组(转染表达ZBED3-AS1的质粒)。WST-1法和Transwell法分析细胞增殖和迁移能力。生物信息学预测ZBED3-AS1的靶基因。qRT-PCR技术分析两组细胞中ZBED3-AS1和靶基因的表达。Western blot法检测靶基因蛋白的表达。结果前列腺癌组织和相应癌旁组织中ZBED3-AS1的表达量分别为0.85±0.26和4.33±0.88(t=18.79,P<0.01)。前列腺癌细胞中ZBED3-AS1的表达低于永生化前列腺上皮细胞(P<0.05),在DU-145中的表达最低(P<0.01)。ZBED3-AS1组中ZBED3-AS1的表达显著增加(P<0.01)。第3天,ZBED3-AS1组细胞OD值低于阴性对照组(P<0.05)。阴性对照组和ZBED3-AS1组DU-145细胞的迁移数分别为189.80±23.07和93.28±13.16(t=3.63,P<0.01)。生物信息学显示,ZBED3-AS1可互补结合miR-512-5p,miR-512-5p可互补结合细胞命运决定因子(DACH1)mRNA。ZBED3-AS1组miR-512-5p的表达显著降低(P<0.01),DACH1 mRNA和蛋白的表达显著增加(P<0.01)。结论前列腺癌组织和细胞中lncRNA ZBED3-AS1的表达水平降低,ZBED3-AS1可能通过吸附miR-512-5p调控DACH1抑制前列腺癌细胞的增殖和迁移。

Purpose To investigate the role of long-chain non-coding RNA(lncRNA)ZBED3-AS1 in the pathogenesis and progression of prostate cancer by regulating miR-512-5p.Methods qRT-PCR was used to analyze the expression of ZBED3-AS1 in prostate cancer tissues and corresponding adjacent tissues in 67 patients with prostate cancer surgery.qRT-PCR was used to analyze the expression of ZBED3-AS1 in prostate cancer cell lines and immortalized prostate epithelial cells.The cells with the lowest expression were divided into a negative control group(transfection negative control plasmid)and a ZBED3-AS1 group(transfected with a plasmid expressing ZBED3-AS1)according to a random number method.The cell proliferation and migration ability were analyzed by the WST-1 method and the Transwell method.Bioinformatics was used to predict the target gene of ZBED3-AS1.qRT-PCR was used to analyze the expression of ZBED3-AS1 and target genes in the two groups of cells.Western blot was used to detect the expression of target gene protein.Results The expression of ZBED3-AS1 in prostate cancer tissues and corresponding adjacent tissues was 0.85±0.26 and 4.33±0.88,respectively(t=18.79,P<0.01).The expression of ZBED3-AS1 in prostate cancer cells was lower than that in immortalized prostate epithelial cells(P<0.05),and the expression in DU-145 was the lowest(P<0.01).The expression of ZBED3-AS1 was significantlyincreased in the ZBED3-AS1 group(P<0.01).From the third day,the OD value of the ZBED3-AS1 group was lower than that of the negative control group(P<0.05).The migration numbers of DU-145 cells in the negative control group and ZBED3-AS1 group were 189.80±23.07 and 93.28±13.16,respectively(t=3.63,P<0.01).Bioinformatics showed that ZBED3-AS1 complemented miR-512-5p,and miR-512-5p complemented the cell fate determinant(DACH1)mRNA.The expression of miR-512-5p in ZBED3-AS1 group was significantly decreased(P<0.01),and the expression of DACH1 mRNA and protein was significantly increased(P<0.01).Conclusion The expression level of lncRNA ZBED3-AS1 is decreased in prostate cancer tissues and cells,and ZBED3-AS1 may inhibit proliferation and migration of prostate cancer cells by adsorption of miR-512-5p and regulation of DACH1 expression.