miR-34a-5p在膀胱癌组织中的表达及其对膀胱癌细胞侵袭迁移的影响和机制

目的:研究miR-34a-5p在膀胱癌组织中的表达及其对膀胱癌细胞侵袭迁移的影响和机制。方法:收集50例膀胱癌组织及对应的癌旁组织,qRT-PCR检测miR-34a-5p表达差异。在膀胱癌细胞中转染miR-34a-5p mimics,qRT-PCR、Transwell小室和Western blot分别检测miR-34a-5p水平、侵袭迁移数目和E-cadherin、N-cadherin、Vimentin蛋白表达水平。靶基因预测软件发现肿瘤蛋白D52（TPD52）可能为miR-34a-5p的靶基因,构建荧光素酶报告载体鉴定靶基因。Western blot检测miR-34a-5p mimics对膀胱癌细胞中TPD52表达的影响。以qRT-PCR检测膀胱癌组织和癌旁组织中TPD52表达变化,分析膀胱癌组织中TPD52表达水平与miR-34a-5p表达水平的相关性。将miR-34a-5p mimics和pcDNA3.1-TPD52共转染至膀胱癌细胞中,按照上述方法检测细胞中TPD52蛋白、侵袭迁移数目及E-cadherin、N-cadherin、Vimentin蛋白表达变化。 结果:miR-34a-5p在膀胱癌组织中表达水平降低,miR-34a-5p mimics提高膀胱癌细胞中miR-34a-5p表达水平,降低膀胱癌细胞侵袭和迁移能力,降低膀胱癌细胞中N-cadherin、Vimentin蛋白表达水平,提高E-cadherin蛋白表达水平。miR-34a-5p靶向调控TPD52表达,并且miR-34a-5p mimics抑制细胞中TPD52蛋白表达。TPD52在膀胱癌组织中高表达,其在膀胱癌组织中的表达水平与miR-34a-5p呈负相关。与共转染miR-34a-5p mimics和pcDNA3.1的细胞比较,miR-34a-5p mimics和pcDNA3.1-TPD52共转染提高膀胱癌细胞中TPD52蛋白表达水平,提高膀胱癌细胞侵袭和迁移能力,促进细胞中N-cadherin、Vimentin蛋白表达,降低E-cadherin蛋白表达水平。结论:miR-34a-5p在膀胱癌组织中低表达,上调miR-34a-5p靶向TPD52抑制膀胱癌细胞侵袭和迁移。

Expression of miR-34a-5p in bladder cancer and its effect on invasion and migration of bladder cancer cells

AIM: To study the expression of miR-34a-5p in bladder cancer and its effect on invasion and migration of bladder cancer cells. METHODS: 50 cases of bladder cancer and corresponding adjacent tissues were collected, the expression of miR-34a-5p was detected by qRT-PCR. The miR-34a-5p mimics were transfected into bladder cancer cells. The levels of miR-34a-5p, the number of invasion and migration, and the expression levels of E-cadherin, N-cadherin and Vimentin were detected by qRT-PCR, Transwell chamber and Western blot, respectively. Target gene prediction software found that TPD52 might be the target gene of miR-34a-5p; luciferase reporter vector was constructed to identify the target gene. The effect of miR-34a-5p mimics on the expression of TPD52 in bladder cancer cells were detected by Western blot. The expression of TPD52 in bladder cancer and adjacent tissues were detected by qRT-PCR, and the correlation between the expression of TPD52 and miR-34a-5p in bladder cancer were analyzed. The miR-34a-5p mimics and pcDNA3.1-TPD52 were co-transfected into bladder cancer cell. TPD52 protein, number of invasion and migration, and expression of E-cadherin, N-cadherin and Vimentin were detected by the above methods. RESULTS:The expression of miR-34a-5p in bladder cancer tissue was decreased. miR-34a-5p mimics increased the expression level of miR-34a-5p in bladder cancer cells, reduced the invasion and migration ability, reduced the expression of N-cadherin and Vimentin, increased the expression of E-cadherin protein. miR-34a-5p regulated the expression of TPD52, moreover, miR-34a-5p mimics inhibited the expression of TPD52 protein in cells. TPD52 was highly expressed in bladder cancer, which was negatively correlated with the expression level of miR-34a-5p in bladder cancer tissue. Compared with co-transfected cells of miR-34a-5p mimics and pcDNA3.1, co-transfection of miR-34a-5p mimics and pcDNA3.1-TPD52 enhanced the expression of TPD52 in bladder cancer cells, increased the invasive and migratory ability of bladder cancer cells, and promoted the expression of N-cadherin and Vimentin, reduced the expression of E-cadherin protein. CONCLUSION: The expression of miR-34a-5p in bladder cancer tissue was low. Upregulation of miR-34a-5p targeting TPD52 inhibits the invasion and migration of bladder cancer cells.