目的:研究lncRNA HOXA11-反义RNA(HOXA11-AS)对膀胱尿路上皮癌J82细胞增殖、侵袭迁移的影响及与微小RNA-515-5p(miR-515-5p)之间的调控关系。方法:qRT-PCR检测膀胱尿路上皮癌组织、癌旁组织和J82细胞中HOXA11-AS和miR-515-5p的转录水平,MTT法和Transwell实验检测J82细胞的增殖、迁移和侵袭能力,Western blot检测细胞中Cyclin D1、p21、p27、MMP-2、MMP-9和MMP-14的表达,双荧光素酶报告系统验证HOXA11-AS和miR-515-5p的结合关系。结果:与正常癌旁组织相比,膀胱尿路上皮癌组织中HOXA11-AS含量显著升高(P<0.05),miR-515-5p的含量显著下降(P<0.05);干扰HOXA11-AS表达可以抑制膀胱尿路上皮癌J82细胞增殖、迁移和侵袭,使Cyclin D1、MMP-2、MMP-9和MMP-14表达下降,p21和p27表达升高;过表达miR-515-5p抑制J82细胞增殖、迁移和侵袭;HOXA11-AS靶向负调控miR-515-5p的表达;抑制miR-515-5p表达可逆转干扰HOXA11-AS表达对J82细胞增殖、迁移和侵袭的作用。结论:LncRNA HOXA11-AS通过靶向miR-515-5p抑制膀胱尿路上皮癌J82细胞增殖、迁移和侵袭。LncRNA HOXA11-AS是膀胱尿路上皮癌的潜在靶点。
Objective: To investigate the effects of lncRNA HOXA11-AS on proliferation,migration and invasion in bladder urothelial carcinoma J82 cells and and the relationship between lncRNA HOXA11-AS and miR-515-5 p. Methods: The transcription levels of lncRNA HOXA11-AS and miR-515-5 p in bladder urothelial carcinoma,adjacent tissues and J82 cells were detected by qRTPCR. The proliferation,migration and invasion abilities of J82 cells were determined by MTT and Transwell assays. The expression levels of Cyclin D1,p21,p27,MMP-2,MMP-9 and MMP-14 in J82 cells were measured by Western blot. The binding relationship between HOXA11-AS and miR-515-5 p was verified by double luciferase reporting system. Results: Compared with normal adjacent tissues,the transcription level of HOXA11-AS in bladder urothelial carcinoma tissues was significantly increased( P<0. 05),and the level of miR-515-5 p was remarkably decreased( P<0. 05). Interfering the expression of HOXA11-AS inhibited the proliferation,migration and invasion of J82 cells,and reduced the expression of Cyclin D1,MMP-2,MMP-9 and MMP-14,and increased the expression of p21 and p27. Over-expression of miR-515-5 p inhibited the proliferation,migration and invasion of J82 cells. LncRNA HOXA11-AS targeted and negatively regulated the transcription of miR-515-5 p. Inhibition of miR-515-5 p reversed the effect of HOXA11-AS interference on proliferation,migration and invasion of J82 cell. Conclusion: LncRNA HOXA11-AS inhibits the proliferation,migration and invasion of bladder urothelial carcinoma J82 cells by targeting miR-515-5 p. LncRNA HOXA11-AS is a potential target for bladder urothelial carcinoma.
Chinese Journal of Immunology