Department of Urology and Reproductive Medicine, Seventh People's Hospital of Shanghai University of TCM, Shanghai 200137, China.
Department of Urology and Reproductive Medicine, Seventh People's Hospital of Shanghai University of TCM, Shanghai 200137, China.
Life Sci. 2020 Oct 15;259:118380. doi: 10.1016/j.lfs.2020.118380. Epub 2020 Sep 6.
Benign prostatic hyperplasia (BPH) is a progressive disease, which severely affects men's health. Here, we sought to analyze the functions and mechanism of action of the tripartite motif protein 52 (TRIM52), a novel prostate basal cell biomarker in BPH.
Immunohistochemistry assay was performed in sectioned human BPH tissues, BPH-1 cells, and prostate RWPE-1 cells, to detect the expressions of TRIM52 and NF-κB. Western blotting and qRT-PCR analyses were conducted to measure the relative protein and mRNA expression levels, respectively. Further, lentiviral transfection was performed in BPH-1 and RWPE-1 cells to study the overexpression and siRNA knockdown of TRIM52. Dual-luciferase reporter assay was applied to evaluate the relationship between NF-κB and TRIM52. Furthermore, CCK-8 assay and flow cytometry were employed to analyze cell proliferation and apoptosis.
TRIM52 and NF-κB levels were elevated in BPH tissues, and TRIM52 expression positively correlated with NF-κB expression. TRIM52 silencing suppressed the growth of BPH-1 cells and decreased the promoter activity of NF-κB. Moreover, the NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC), suppressed TRIM52-induced proliferation of RWPE-1 cells and inhibited NF-κB promoter activity in oeTRIM52 transfected RWPE-1 cells. Silencing TRIM52 also inhibited TRAF2 ubiquitination in BPH-1 cells. Further, NF-κB promoter activity in siNC transfected cells was enhanced by the recombinant protein TNF-α and inhibited by siTRIM52.
TRIM52 accelerated the growth of BPH-1 cells by upregulating NF-κB, and TRIM52 could promote TRAF2 ubiquitination. These findings might contribute to the understanding of the biological functions and action mechanisms of TRIM52 in BPH.
良性前列腺增生（BPH）是一种进行性疾病，严重影响男性健康。在这里，我们试图分析三结构域蛋白 52（TRIM52）在 BPH 中的作用和作用机制，TRIM52 是一种新的前列腺基底细胞生物标志物。
对 BPH 组织、BPH-1 细胞和前列腺 RWPE-1 细胞的切片进行免疫组织化学检测，以检测 TRIM52 和 NF-κB 的表达。Western blot 和 qRT-PCR 分析分别用于测量相对蛋白和 mRNA 表达水平。进一步在 BPH-1 和 RWPE-1 细胞中进行慢病毒转染，以研究 TRIM52 的过表达和 siRNA 敲低。双荧光素酶报告基因检测用于评估 NF-κB 和 TRIM52 之间的关系。进一步采用 CCK-8 测定和流式细胞术分析细胞增殖和凋亡。
TRIM52 和 NF-κB 水平在 BPH 组织中升高，TRIM52 表达与 NF-κB 表达呈正相关。TRIM52 沉默抑制 BPH-1 细胞的生长，并降低 NF-κB 的启动子活性。此外，NF-κB 抑制剂吡咯烷二硫代氨基甲酸酯（PDTC）抑制了 oeTRIM52 转染的 RWPE-1 细胞中 TRIM52 诱导的增殖，并抑制了 NF-κB 启动子活性。沉默 TRIM52 还抑制了 BPH-1 细胞中 TRAF2 的泛素化。进一步，重组蛋白 TNF-α增强了 siNC 转染细胞中 NF-κB 启动子活性，而 siTRIM52 抑制了 NF-κB 启动子活性。
TRIM52 通过上调 NF-κB 加速 BPH-1 细胞的生长，TRIM52 可以促进 TRAF2 泛素化。这些发现可能有助于理解 TRIM52 在 BPH 中的生物学功能和作用机制。
