雷公藤红素通过上调 microRNA-218 抑制良性前列腺上皮细胞活力和迁移并诱导细胞凋亡。
Triptolide inhibits benign prostatic epithelium viability and migration and induces apoptosis via upregulation of microRNA-218.
机构信息
1 Department of Urinary Surgery, People's Hospital of Rizhao, Rizhao, China.
2 Department of Urinary Surgery, Affiliated Hospital of Taishan Medical University, Taian, China.
出版信息
Int J Immunopathol Pharmacol. 2018 Mar-Dec;32:2058738418812349. doi: 10.1177/2058738418812349.
Benign prostatic hypertrophy (BPH) has become a troublesome disease for elder men. Triptolide (TPL) has been reported to be a potential anticancer agent. However, the potential effects of TPL on BPH have not been shown out. BPH-1 cells were treated with different concentrations of TPL and/or transfected with microRNA-218 (miR-218) inhibitor, pc-survivin, sh-survivin, or their corresponding controls (NC). Thereafter, cell viability was determined by CCK-8 assay. Cell migration was accessed by modified two-chamber migration assay. Cell apoptosis was checked by propidium iodide (PI) and fluorescein isothiocyanate (FITC)-conjugated Annexin V staining. In addition, messenger RNA (mRNA) and protein levels were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis, respectively. BPH-1 cell viability and migration were significantly decreased, while cell apoptosis and expression of miR-218 were statistically enhanced by TPL ( P < 0.05 or P < 0.01). However, downregulation of miR-218 increased cell viability and migration, while decreased cell apoptosis compared with the negative control group ( P < 0.05 or P < 0.01). Furthermore, the expression of cell cycle-related proteins and cell apoptosis-related proteins were also led to the opposite results with NC. In addition, we found that miR-218 negatively regulated the expression of survivin ( P < 0.01) and suppression of survivin significantly enhanced cell apoptosis ( P < 0.01). Moreover, the results demonstrated that TPL could inactivate mammalian target of rapamycin (mTOR) pathway, while inhibition of miR-218 alleviated the effects. TPL inhibits viability and migration of BPH-1 cells and induces cell apoptosis and also inactivates mTOR signal pathway via upregulation of miR-218. This study provides evidence for the further studies representing triptolide as a potential agent in the treatment of human BPH.
良性前列腺增生（BPH）已成为老年男性的一种麻烦疾病。雷公藤内酯醇（TPL）已被报道为一种潜在的抗癌药物。然而，TPL 对 BPH 的潜在影响尚未显现。用不同浓度的 TPL 处理 BPH-1 细胞和/或用 microRNA-218（miR-218）抑制剂、pc-survivin、sh-survivin 或其相应的对照物（NC）转染 BPH-1 细胞。此后，通过 CCK-8 测定法测定细胞活力。通过改良的双层迁移测定法评估细胞迁移。通过碘化丙啶（PI）和荧光素异硫氰酸酯（FITC）缀合的 Annexin V 染色检查细胞凋亡。此外，使用定量实时聚合酶链反应（qRT-PCR）和 Western blot 分析分别检测信使 RNA（mRNA）和蛋白质水平。TPL 显著降低了 BPH-1 细胞活力和迁移，而 miR-218 的表达则显著增强了细胞凋亡（P<0.05 或 P<0.01）。然而，与阴性对照组相比，下调 miR-218 增加了细胞活力和迁移，同时降低了细胞凋亡（P<0.05 或 P<0.01）。此外，细胞周期相关蛋白和细胞凋亡相关蛋白的表达也与 NC 相反。此外，我们发现 miR-218 负调控 survivin 的表达（P<0.01），抑制 survivin 显著增强细胞凋亡（P<0.01）。此外，结果表明 TPL 可以使哺乳动物雷帕霉素靶蛋白（mTOR）途径失活，而抑制 miR-218 可减轻这种作用。TPL 通过上调 miR-218 抑制 BPH-1 细胞活力和迁移，诱导细胞凋亡，并使 mTOR 信号通路失活。这项研究为进一步研究雷公藤内酯醇作为治疗人类 BPH 的潜在药物提供了证据。
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