CCND1, NOP14 and DNMT3B are involved in miR-502-5p-mediated inhibition of cell migration and proliferation in bladder cancer
CCND1, NOP14 and DNMT3B are involved in miR-502-5p-mediated inhibition of cell migration and proliferation in bladder cancer
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CCND1、NOP14 和 DNMT3B 参与 miR-502-5p 介导的膀胱癌细胞迁移和增殖抑制
DOI：
10.1111/cpr.12751
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发表时间：
2020-01-23
影响因子：
8.5
通讯作者：
Xie, Liping
中科院分区：
文献类型：
--
作者：
Ying, Yufan;Li, Jiangfeng;Xie, Liping
目的miR-502- 5 p的下调已成为几种癌症肿瘤进展的关键因素。在此，我们阐明了miR-502- 5 p在膀胱癌中的作用。材料与方法采用RT-qPCR方法检测miR-502- 5 p在膀胱癌中的表达。DNA甲基化分析表明，表观遗传学机制可能参与了miR-502- 5 p的下调。采用创伤愈合实验、transwell实验、集落形成实验、CCK 8实验和流式细胞术分析miR-502- 5 p在膀胱癌细胞中的作用。Western blot检测相关基因的蛋白水平。此外，还根据需要进行双荧光素酶报告基因测定、体内成瘤测定和免疫组织化学染色。结果miR-502- 5 p在BCa中表达下调。同时，CpG岛的高甲基化参与了miR-502- 5 p的下调。在功能上，miR-502- 5 p的过表达在体外抑制细胞增殖和迁移，并在体内抑制肿瘤生长。CCND 1、DNMT 3B和N 0 P14被鉴定为miR-502- 5 p的直接靶点。有趣的是，DNMT 3B和miR-502- 5 p在膀胱癌的调控中建立了正反馈回路。此外，拯救实验进一步验证了miR-502- 5 p与其靶点之间的直接分子相互作用。结论miR-502- 5 p介导的调控网络在膀胱癌的发生发展中起着重要作用，该调控网络可能有助于开发更有效的膀胱癌治疗方法。
Objectives Downregulation of miR-502-5p has emerged as a critical factor in tumour progression in several cancers. Herein, we elucidated the role of miR-502-5p in bladder cancer. Materials and methods RT-qPCR was performed to examine the expression of miR-502-5p in bladder cancer. And DNA methylation analysis showed that epigenetic mechanisms may contribute to the downregulation of miR-502-5p. Then, wound-healing assay, transwell assay, colony formation assay, CCK8 assay and flow cytometry analysis were applied to evaluate the function of miR-502-5p in bladder cancer cell lines. Western blot was conducted to measure the protein levels of related genes. Furthermore, dual-luciferase reporter assay, in vivo tumorigenesis assay and immunohistochemical staining were also conducted as needed. Results MiR-502-5p is frequently downregulated in BCa. Meanwhile, hypermethylation of CpG islands contributes to the downregulation of miR-502-5p. Functionally, overexpression of miR-502-5p inhibited cell proliferation and migration in vitro and repressed tumour growth in vivo. CCND1, DNMT3B and NOP14 were identified as direct targets of miR-502-5p. Interestingly, DNMT3B and miR-502-5p established a positive feedback loop in the regulation of bladder cancer. In addition, rescue experiments further validated the direct molecular interaction between miR-502-5p and its targets. Conclusions Our study proposed and demonstrated that the miR-502-5p-mediated regulatory network is critical in bladder cancer; this network may be useful in the development of more effective therapies against bladder cancer.