miR-372-3p 在膀胱癌组织中的表达及其对膀胱癌细胞增殖、迁移与侵袭的影响

目的：检测miR-372-3p 在膀胱癌组织和转染miR-372-3p mimics 膀胱癌细胞中的表达，研究miR-372-3p 对膀胱癌5637 和T24 细胞增殖、迁移与侵袭的影响。方法：采集2016 年3 月至2017 年1 月武汉中心医院收治的6 例膀胱癌患者癌及癌旁组织（距离肿瘤边缘>5 cm），qPCR检测膀胱癌及癌旁组织miR-372-3p 表达。采用脂质体转染法将miR-372-3p mimics 或者miRNC转入膀胱癌5637 和T24 细胞。用qPCR和Western blotting 检测转染miR-372-3p mimics 和miR-NC膀胱癌细胞miR-372-3p 和ATAD2 mRNA和蛋白E-cadherin、N-cadherin 蛋白的表达，流式细胞术检测细胞周期分布，MTT法和集落形成实验检测细胞增殖和集落形成能力，划痕实验和Transwell 侵袭实验检测细胞的迁移和侵袭能力。结果：膀胱癌组织miR-372-3p mRNA的表达显著低于癌旁组织（0.65±0.56 vs 1.76±0.34，P<0.01）。和转染miR-NC膀胱癌细胞相比，转染miR-372-3p mimics 膀胱癌细胞的miR-372-3p mRNA表达显著增加，ATAD2 mRNA和蛋白的表达显著降低，E-cadherin 蛋白表达上调，N-cadherin 蛋白表达下调，细胞周期明显阻滞，细胞集落形成和增殖能力显著降低，细胞迁移数和侵袭数减少。结论：miR-372-3p 的低表达可能与膀胱癌的发生发展有关，其通过靶向调控ATAD2 抑制膀胱癌细胞的增殖、迁移和侵袭能力，可能成为膀胱癌生物治疗的新靶标。

Expression of miR- 372- 3p in the tissues of bladder carcinoma and its effect on proliferation, migration and invasion of the bladder cancer cells

Objective: To detect expression of miR-372-3p in the bladder cancer cell transfected with miR-372-3p mimics and to explore effect of miR-372-3p on proliferation, migration and invasion of the bladder cancer 5637 and T24 cell lines. Methods: Using lipofection,miR-372-3p mimics or miR-NC was transfected into the bladder cancer 5637 and T24 cells. Expressions of miR-372-3p mRNA and ATAD2 mRNA, expressions of ADAD2, E-cadherin and N-cadherin proteins, cell cycle distribution, viability of cell, cell proliferation ability, cell migration ability and cell invasion ability in the bladder cancer cells transfected with miR-372-3p mimics and miR-NC were respectively detected by qPCR, Western blotting assay, flow cytometry, MTT assay, colony forming test, scratch assay and Transwell assay. Results: Comparing with the bladder cancer cell transfected with miR-NC, expression of miR-372-3p mRNA significantly increased, expressions of ATAD2 mRNA and protein obviously decreased, expression of E-cadherin protein raised, expression of N-cadherin protein came down, cell cycle obviously arrested, abilities of cell viability and proliferation markedly reduced as well as cell numbers of migration and invasion reduced, in the bladder cancer cell transfected with miR-372-3p mimics. Conclusion: Low expression of miR-372-3p might be associated with occurrence and development of the bladder carcinoma. miR-372-3p could inhibit abilities of proliferation,migration and invasion of the bladder cancer cell through targeting control. It could be a novel target in biotherapy of the bladder carcinoma.