Department of Applied Biology, CSIR-Indian Institute of Chemical Technology (CSIR-IICT), Hyderabad, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, India.
Prostate. 2023 Jul;83(10):936-949. doi: 10.1002/pros.24535. Epub 2023 Apr 17.
Prostate cancer (PCa) is the leading cause of cancer related deaths in men, often androgen deprivation therapy (ADT) leads to the progression of androgen independent PCa (AIPC) which further leads to Neuroendocrine PCa (NEPC). Identifying the molecular mechanisms which navigate the neuroendocrine differentiation (NED) of PCa cells is clinically relevant. It has been suggested that the micro RNAs (miRNAs) play an important role in the regulation of intrinsic mechanisms relevant to tumor progression, resistance as a result leads to poor prognosis. miR-147b has been transpiring as one of the deregulated miRNAs associated with the occurrence of multiple cancers. The present study has studied the role of miRNA-147b in inducing NEPC.
To investigate the functional role of miR-147b in NEPC, we have expressed miRNA mimics or inhibitors in PCa cells and monitored the progression of NEPC along with PCa cell proliferation and survival. The molecular mechanism miRNA-147b follows was studied using western blot and reverse transcription polymerase chain analysis. miRNA target prediction using bioinformatics tools followed by target validation using luciferase reporter assays was performed.
In the present study, we found that miR-147b is highly expressed in AIPC cell lines in particular neuroendocrine cells NCI-H660 and NE-LNCaP derived from LNCaP. Mechanistic studies revealed that overexpression of miR-147b or miRNA mimics induced NED in LNCaP cells in in-vitro while its inhibitor reversed the NE features (increased NE markers and reduced prostate specific antigen) of PC3, NCI-H660 and NE-LNCaP cells. In addition, miR-147b reduced the proliferation rate of LNCaP cells via elevated p27kip1 and lowered cyclin D1 for promoting differentiation. In reporter assays, we have identified ribosomal protein S15A (RPS15A) is a direct target of miRNA-147b and RPS15A expression was negatively regulated by miR-147b in PCa cells. Furthermore, we also report that RPS15A is downregulated in NEPC cells and its expression is inversely correlated with NE markers.
Targeting the miR-147b - RPS15A axis may overcome the progression of NEPC and serve as a novel therapeutic target to attenuate NED progression of PCa.
前列腺癌（PCa）是男性癌症相关死亡的主要原因，通常雄激素剥夺疗法（ADT）导致雄激素非依赖性 PCa（AIPC）的进展，进而导致神经内分泌 PCa（NEPC）。鉴定引导 PCa 细胞神经内分泌分化（NED）的分子机制具有临床相关性。已经表明 microRNAs（miRNAs）在调节与肿瘤进展、耐药相关的内在机制方面发挥着重要作用，导致预后不良。miR-147b 已被证明是与多种癌症发生相关的失调 miRNAs 之一。本研究研究了 miR-147b 在诱导 NEPC 中的作用。
为了研究 miR-147b 在 NEPC 中的功能作用，我们在 PCa 细胞中表达 miRNA 模拟物或抑制剂，并监测 NEPC 的进展以及 PCa 细胞的增殖和存活。使用 Western blot 和逆转录聚合酶链分析研究了 miRNA-147b 遵循的分子机制。使用生物信息学工具进行 miRNA 靶标预测，然后使用荧光素酶报告基因检测进行靶标验证。
在本研究中，我们发现 miR-147b 在 AIPC 细胞系中高度表达，特别是神经内分泌细胞 NCI-H660 和源自 LNCaP 的 NE-LNCaP。机制研究表明，miR-147b 的过表达或 miRNA 模拟物在体外诱导 LNCaP 细胞的 NED，而其抑制剂则逆转了 PC3、NCI-H660 和 NE-LNCaP 细胞的 NE 特征（增加 NE 标志物和降低前列腺特异性抗原）。此外，miR-147b 通过上调 p27kip1 和降低 cyclin D1 来降低 LNCaP 细胞的增殖率，从而促进分化。在报告基因检测中，我们已经确定核糖体蛋白 S15A（RPS15A）是 miRNA-147b 的直接靶标，并且在 PCa 细胞中，RPS15A 的表达受 miR-147b 的负调控。此外，我们还报告说，RPS15A 在 NEPC 细胞中下调，其表达与 NE 标志物呈负相关。
靶向 miR-147b-RPS15A 轴可能克服 NEPC 的进展，并作为一种新的治疗靶点来减弱 PCa 的 NED 进展。
