miRNAs 203a/210-3p/5001-5p 调节雄激素/雄激素受体/YAP 诱导的前列腺癌细胞迁移。

The miRNAs 203a/210-3p/5001-5p regulate the androgen/androgen receptor/YAP-induced migration in prostate cancer cells.

机构信息

Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan, Taiwan.

Ph.D. Program for Cancer Molecular Biology and Drug Discovery, Taipei Medical University, Taipei, Taiwan.

出版信息

Cancer Med. 2024 Aug;13(16):e70106. doi: 10.1002/cam4.70106.

DOI:10.1002/cam4.70106

PMID:39149855

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11327718/

Abstract

BACKGROUND

Prostate cancer (PCa) patients with elevated level of androgen receptor (AR) correlate with higher metastatic incidence. Protein expression of AR and its target gene prostate-specific antigen (PSA) are elevated in metastatic prostate tumors as compared to organ-confined tumors. Androgen treatment or elevation of AR promotes metastasis of PCa in cell culture and murine model. However, under androgen depleted condition, AR suppressed cell mobility and invasiveness of PCa cells. Androgen deprivation therapy in PCa patients is associated with higher risk of cancer metastasis. We therefore investigated the dual roles of AR and miRNAs on PCa metastasis.

METHODS

The PC-3 (PC-3 cells re-expressing AR) and LNCaP cells were used as PCa cell model. Transwell migration and invasion assay, wound-healing assay, zebrafish xenotransplantation assay, and zebrafish vascular exit assay were used to investigate the role of AR and androgen on PCa metastasis. Micro-Western Array, co-immunoprecipitation and Immunofluorescence were applied to dissect the molecular mechanism lying underneath. The miRNA array, miRNA inhibitors or plasmid, and chromatin immunoprecipitation assay were used to study the role of miRNAs on PCa metastasis.

RESULTS

In the absence of androgen, AR repressed the migration and invasion of PCa cells. When androgen was present, AR stimulated the migration and invasion of PCa cells both in vitro and in zebrafish xenotransplantation model. Androgen increased phospho-AR Ser81 and yes-associated protein 1 (YAP), decreased phospho-YAP Ser217, and altered epithelial-mesenchymal transition (EMT) proteins in PCa cells. Co-IP assay demonstrated that androgen augmented the interaction between YAP and AR in nucleus. Knockdown of YAP or treatment with YAP inhibitor abolished the androgen-induced migration and invasion of PCa cells, while overexpression of YAP showed opposite effects. The miRNA array revealed that androgen decreased hsa-miR-5001-5p but increased hsa-miR-203a and hsa-miR-210-3p in PC-3AR cells but not PC-3 cells. Treatment with inhibitors targeting hsa-miR-203a/hsa-miR-210-3p, or overexpression of hsa-miR-5001-5p decreased YAP expression as well as suppressed the androgen-induced migration and invasion of PCa cells. Chromatin immunoprecipitation (ChIP) assay demonstrated that AR binds with promoter region of has-miR-210-3p in the presence of androgen.

CONCLUSIONS

Our observations indicated that miRNAs 203a/210-3p/5001-5p regulate the androgen/AR/YAP-induced PCa metastasis.

摘要

背景

雄激素受体（AR）水平升高的前列腺癌（PCa）患者与更高的转移性发生率相关。与器官受限肿瘤相比，转移性前列腺肿瘤中 AR 的蛋白表达及其靶基因前列腺特异性抗原（PSA）升高。在细胞培养和小鼠模型中，雄激素治疗或 AR 升高会促进 PCa 的转移。然而，在雄激素耗尽的情况下，AR 抑制了 PCa 细胞的迁移和侵袭。在 PCa 患者中进行雄激素剥夺疗法会增加癌症转移的风险。因此，我们研究了 AR 和 miRNAs 对 PCa 转移的双重作用。

方法

PC-3（重新表达 AR 的 PC-3 细胞）和 LNCaP 细胞被用作 PCa 细胞模型。使用 Transwell 迁移和侵袭测定、划痕愈合测定、斑马鱼异种移植测定和斑马鱼血管出口测定来研究 AR 和雄激素对 PCa 转移的作用。应用微量西方阵列、免疫共沉淀和免疫荧光来剖析潜在的分子机制。miRNA 阵列、miRNA 抑制剂或质粒和染色质免疫沉淀测定用于研究 miRNAs 对 PCa 转移的作用。

结果

在没有雄激素的情况下，AR 抑制了 PCa 细胞的迁移和侵袭。当存在雄激素时，AR 在体外和斑马鱼异种移植模型中均刺激 PCa 细胞的迁移和侵袭。雄激素增加了磷酸化 AR Ser81 和 YAP，减少了磷酸化 YAP Ser217，并改变了 PCa 细胞中的上皮-间充质转化（EMT）蛋白。免疫共沉淀测定表明，雄激素增强了细胞核中 YAP 和 AR 之间的相互作用。敲低 YAP 或用 YAP 抑制剂处理消除了雄激素诱导的 PCa 细胞迁移和侵袭，而过表达 YAP 则显示出相反的效果。miRNA 阵列显示，雄激素降低了 PC-3AR 细胞中的 hsa-miR-5001-5p，但增加了 PC-3 细胞中的 hsa-miR-203a 和 hsa-miR-210-3p。用靶向 hsa-miR-203a/hsa-miR-210-3p 的抑制剂处理或过表达 hsa-miR-5001-5p 会降低 YAP 表达并抑制雄激素诱导的 PCa 细胞迁移和侵袭。染色质免疫沉淀（ChIP）测定表明，在存在雄激素的情况下，AR 与 hsa-miR-210-3p 的启动子区域结合。