Department of Urology, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Eur Rev Med Pharmacol Sci. 2019 Jan;23(2):530-538. doi: 10.26355/eurrev_201901_16865.
The aim of this study was to investigate the inhibitory role of microRNA-1299 (miR-1299) in prostate cancer, and to explore the possible underlying mechanism.
The expression of miR-1299 in 35 PCa tissues and para-carcinoma tissues, as well as PCa cell lines (PC-3) and prostatic epithelial cell line (RWPE-1), was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Then, we explored the possible targets of miR-1299 by searching online databases. NIMA-related kinase 2 (NEK2) was identified as a direct target gene of miR-1299. Subsequently, qRT-PCR, Western blot (WB), and luciferase reporter gene assay were used to further verify the correlation between miR-1299 and NEK2. To better characterize the role of miR-1299 and NEK2 in PCa, we conducted functional experiments (MTT, flow cytometry, scratch-wound, and transwell assay) by transfecting PC-3 cells with miR-1299 mimics and si-NEK2 in different groups.
The expression level of miR-1299 in PCa tissues was significantly lower than that of para-carcinoma tissues. Meanwhile, the expression of miR-1299 in PC-3 cells was also significantly downregulated when compared with RWPE-1 cells. Subsequent qRT-PCR, WB, and luciferase reporter gene assay verified that miR-1299 transcriptionally repressed NEK2 by interacting with the essential binding sequence in 3'-UTR. Also, functional experiments demonstrated that decreased expression of NEK2 resulting from miR-1299 up-regulation could remarkably inhibit the proliferation, invasion, and migration of PCa cells.
Our study indicated that miR-1299 was a novel suppressor in PCa through its negative regulation of NEK2. Moreover, our findings revealed that miR-1299/NEK2 axis might be a potential therapeutic target for the treatment of PCa.
本研究旨在探讨 microRNA-1299（miR-1299）在前列腺癌中的抑制作用，并探讨其可能的潜在机制。
通过定量实时聚合酶链反应（qRT-PCR）检测 35 例前列腺癌组织和癌旁组织、前列腺癌细胞系（PC-3）和前列腺上皮细胞系（RWPE-1）中 miR-1299 的表达。然后，我们通过在线数据库搜索来探索 miR-1299 的可能靶点。NIMA 相关激酶 2（NEK2）被鉴定为 miR-1299 的直接靶基因。随后，使用 qRT-PCR、Western blot（WB）和荧光素酶报告基因检测进一步验证 miR-1299 与 NEK2 之间的相关性。为了更好地描述 miR-1299 和 NEK2 在前列腺癌中的作用，我们通过转染 miR-1299 模拟物和不同组的 si-NEK2 对 PC-3 细胞进行功能实验（MTT、流式细胞术、划痕愈合和 Transwell 测定）。
miR-1299 在前列腺癌组织中的表达水平明显低于癌旁组织。同时，与 RWPE-1 细胞相比，PC-3 细胞中的 miR-1299 表达也明显下调。随后的 qRT-PCR、WB 和荧光素酶报告基因检测证实，miR-1299 通过与 3'-UTR 中的必需结合序列相互作用，转录抑制 NEK2。此外，功能实验表明，miR-1299 上调导致 NEK2 表达降低可显著抑制前列腺癌细胞的增殖、侵袭和迁移。
本研究表明，miR-1299 通过负调控 NEK2 成为前列腺癌中的一种新型抑制因子。此外，我们的研究结果表明，miR-1299/NEK2 轴可能是治疗前列腺癌的潜在治疗靶点。
