Department of Urology, The First Affiliated Hospital of Bengbu Medical College, Bengbu City, 233000, Anhui Province, China.
Cell Biochem Biophys. 2024 Jun;82(2):1567-1578. doi: 10.1007/s12013-024-01308-7. Epub 2024 May 27.
Prostate cancer (PCa) is a prevalent male malignancy globally. Tripartite motif 47 (TRIM47) has been reported to be associated with PCa. However, how TRIM47 acts on PCa is still incompletely understood. Here, we explored the biological roles of TRIM47 in PCa cells and investigated its potential regulatory mechanism. TRIM47 expression in PCa cells was detected by qRT-PCR and western blot. After TRIM47 silencing, the viability of PCa cells was measured using CCK-8 method. Flow cytometry was employed to estimate cell cycle. Cell apoptotic level was subjected to appraisement with TUNEL assay. Additionally, wound healing- and transwell assays were adopted for evaluation of migration and invasion of PCa cells. Moreover, the Biogrid database and HDOCK SERVER predicated that TRIM47 could interact with mouse double minute 2 (MDM2), which was detected using the Co-immunoprecipitation (co-IP) assay and glutathione S-transferase (GST) pull-down assay. The expression of proteins in MDM2/p53 signaling was detected by western blot analysis. Results indicated that TRIM47 expression was highly expressed in PCa cells. TRIM47 knockdown inhibited PCa proliferation and cell cycle whereas promoted cell apoptosis. Besides, TRIM47 knockdown significantly inhibited the migration and invasion of PCa cells. In addition, TRIM47 was proved to bind to MDM2 and regulated MDM2/p53 expression. Importantly, MDM2 overexpression counteracted the impacts of TRIM47 knockdown on cell viability, cell cycle, apoptosis, migration and invasion by regulating the MDM2/p53 pathway. Collectively, our results suggested that TRIM47 silencing inhibits the malignant biological behaviors of prostate cancer cells by regulating MDM2/p53 signaling, which may provide a novel therapeutic target for PCa treatment.
前列腺癌（PCa）是全球普遍存在的男性恶性肿瘤。三结构域蛋白 47（TRIM47）已被报道与 PCa 相关。然而，TRIM47 如何作用于 PCa 仍不完全清楚。在这里，我们探讨了 TRIM47 在 PCa 细胞中的生物学作用，并研究了其潜在的调节机制。通过 qRT-PCR 和 Western blot 检测 PCa 细胞中 TRIM47 的表达。沉默 TRIM47 后，使用 CCK-8 法测量 PCa 细胞的活力。通过流式细胞术评估细胞周期。采用 TUNEL 检测评估细胞凋亡水平。此外，采用划痕愈合和 Transwell 测定评估 PCa 细胞的迁移和侵袭。此外，使用 Biogrid 数据库和 HDOCK SERVER 预测 TRIM47 可以与鼠双微体 2（MDM2）相互作用，并用免疫共沉淀（co-IP）和谷胱甘肽 S-转移酶（GST）下拉实验检测。Western blot 分析检测 MDM2/p53 信号通路中蛋白的表达。结果表明，TRIM47 在 PCa 细胞中高表达。TRIM47 敲低抑制 PCa 增殖和细胞周期，而促进细胞凋亡。此外，TRIM47 敲低显著抑制 PCa 细胞的迁移和侵袭。此外，TRIM47 被证明与 MDM2 结合，并调节 MDM2/p53 表达。重要的是，MDM2 过表达通过调节 MDM2/p53 通路逆转了 TRIM47 敲低对细胞活力、细胞周期、凋亡、迁移和侵袭的影响。总之，我们的研究结果表明，TRIM47 沉默通过调节 MDM2/p53 信号通路抑制前列腺癌细胞的恶性生物学行为，这可能为 PCa 的治疗提供新的治疗靶点。
