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

doi:10.12092/j.issn.1009-2501.2019.04.007

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (4): 403-410.doi: 10.12092/j.issn.1009-2501.2019.04.007

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

李鹏¹，杨荣华¹，张明华¹，肖鑫¹，汤建儿²

¹湖州市中心医院泌尿外科，湖州 313000，浙江； ²湖州市第一人民医院，湖州师范大学附属第一医院，湖州 313000，浙江

收稿日期:2018-12-03 修回日期:2019-04-09 出版日期:2019-04-26 发布日期:2019-05-01
作者简介:李鹏，男，硕士，副主任医师，研究方向：泌尿系肿瘤微创治疗。 E-mail：hezha1477@163.com
基金资助:
浙江省基础公益研究计划项目（LGF19H160005）

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

LI Peng¹, YANG Ronghua ¹, ZHANG Minghua ¹, XIAO Xin ¹, TANG Jianer ²

¹Department of Urology, Huzhou Central Hospital, Huzhou 313000, Zhejiang, China; ²Huzhou First People's Hospital, the First Affiliated Hospital of Huzhou Normal University, Huzhou 313000, Zhejiang, China

Received:2018-12-03 Revised:2019-04-09 Online:2019-04-26 Published:2019-05-01

摘要/Abstract

摘要：

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。

关键词: 膀胱癌, miR-34a-5p, 侵袭, 肿瘤蛋白D52

Abstract:

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.

Key words: bladder cancer, miR-34a-5p, invasion, TPD52

中图分类号:

R737.14

李鹏，杨荣华，张明华，肖鑫，汤建儿. 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[J]. 中国临床药理学与治疗学, 2019, 24(4): 403-410.

LI Peng, YANG Ronghua, ZHANG Minghua, XIAO Xin, TANG Jianer. Expression of miR-34a-5p in bladder cancer and its effect on invasion and migration of bladder cancer cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(4): 403-410.

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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

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

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