miR-1199-5p 的上调与良性前列腺增生中 2 型 5-α 还原酶表达降低有关。

Upregulation of mir-1199-5p is associated with reduced type 2 5-α reductase expression in benign prostatic hyperplasia.

机构信息

Beijing Shijitan Hospital, Capital Medical University, 100038, Beijing, China.

Department of Urology, Beijing Chaoyang Hospital, Capital Medical University, 100016, Beijing, China.

出版信息

BMC Urol. 2022 Nov 7;22(1):172. doi: 10.1186/s12894-022-01121-5.

DOI:10.1186/s12894-022-01121-5

PMID:36344974

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9639318/

Abstract

BACKGROUND

5-α reductase inhibitors (5-ARIs) are first-line drugs for managing benign prostatic hyperplasia (BPH). Unfortunately, some patients do not respond to 5-ARI therapy and may even show worsening symptoms. The decreased expression of steroid 5-α reductase type 2(SRD5A2) in BPH tissues may explain the failure of 5-ARI therapy, however, the mechanisms underlying SRD5A2 decreased remained unelucidated.

OBJECTIVES

To investigate microRNA-mediated regulation of the expression of SRD5A2 resulting in 5-ARI therapy failure.

MATERIALS AND METHODS

The expression of SRD5A2 and microRNAs in BPH tissues and prostate cells were detected by immunohistochemistry, western blotting, and quantitative real-time PCR. Dual-luciferase reporter assay was performed to confirm that microRNA directly combine to SRD5A2 mRNA. The apoptosis of prostatic cells was detected by flow cytometry.

RESULTS

SRD5A2 expression was variable; it was negative, weak, and strong in 13.6%, 28.8%, and 57.6% of BPH tissues respectively. The normal human prostatic epithelial cell line RWPE-1 strongly expressed SRD5A2, whereas the immortalized human prostatic epithelial cell line BPH-1 weakly expressed SRD5A2. miR-1199-5p expression was remarkably higher in BPH-1 than in RWPE-1 cells(P<0.001), and miR-1199-5p expression was significantly upregulated in BPH tissues with negative SRD5A2 expression than those with positive SRD5A2 expression. Transfection of miR-1199-5p mimics in RWPE-1 cells led to a marked decrease in SRD5A2 expression, whereas miR-1199-5p inhibitor increased SRD5A2 expression in BPH-1 cells. Dual-luciferase reporter assay showed that miR-1199-5p could bind the 3'untranslated region of SRD5A2 mRNA. miR-1199-5p also decreased the RWPE-1 sensibility to finasteride, an inhibitor of SRD5A2.

CONCLUSION

Our results show that SRD5A2 expression varies in BPH tissues and miR-1199-5p might be one of the several factors contributing to differential SRD5A2 expression in BPH patients.

摘要

背景

5-α 还原酶抑制剂（5-ARIs）是治疗良性前列腺增生（BPH）的一线药物。不幸的是，一些患者对 5-ARI 治疗无反应，甚至症状恶化。BPH 组织中类固醇 5-α 还原酶 2（SRD5A2）的表达降低可能解释了 5-ARI 治疗的失败，但 SRD5A2 降低的机制仍不清楚。

目的

研究 microRNA 介导的 SRD5A2 表达调控导致 5-ARI 治疗失败的机制。

材料与方法

通过免疫组织化学、Western blot 和实时定量 PCR 检测 BPH 组织和前列腺细胞中 SRD5A2 和 microRNA 的表达。双荧光素酶报告基因实验证实 microRNA 可直接与 SRD5A2 mRNA 结合。通过流式细胞术检测前列腺细胞的凋亡。

结果

SRD5A2 的表达存在差异；在 13.6%、28.8%和 57.6%的 BPH 组织中，SRD5A2 的表达分别为阴性、弱阳性和强阳性。正常的人前列腺上皮细胞系 RWPE-1 强烈表达 SRD5A2，而永生化的人前列腺上皮细胞系 BPH-1 则弱表达 SRD5A2。miR-1199-5p 在 BPH-1 细胞中的表达明显高于 RWPE-1 细胞（P<0.001），且在 SRD5A2 表达阴性的 BPH 组织中 miR-1199-5p 的表达明显上调。在 RWPE-1 细胞中转染 miR-1199-5p 模拟物可导致 SRD5A2 表达显著降低，而 miR-1199-5p 抑制剂则可增加 BPH-1 细胞中 SRD5A2 的表达。双荧光素酶报告基因实验表明，miR-1199-5p 可与 SRD5A2 mRNA 的 3'UTR 结合。miR-1199-5p 还降低了抑制剂 finasteride 对 RWPE-1 细胞的敏感性，finasteride 是 SRD5A2 的抑制剂。