Department of Urology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China.
Eur Rev Med Pharmacol Sci. 2018 Jun;22(12):3734-3741. doi: 10.26355/eurrev_201806_15253.
Extra-cellular signal regulated kinase (ERK)/mitogen activated protein kinase (MAPK) signaling pathway is widely involved in cell proliferation, apoptosis, and drug resistance. MAPK kinase 1 (MEK1) is the upstream protein kinase of ERK that can activate ERK/MAPK signaling pathway. microRNA 203 (MiR-203) down-regulation is found to be associated with prostate cancer pathogenesis. Bioinformatics analysis showed the complementary targeted relationship between miR-203 and the 3'-UTR of MEK1 mRNA. This study explored the role of miR-203 in regulating prostate cancer cell proliferation, apoptosis, and ADM resistance through affecting MEK1 expression.
Dual luciferase assay confirmed the targeted relationship between miR-203 and MEK1. MiR-203, MEK1, p-ERK1/2, and B cell lymphoma 2 (Bcl-2) expressions were compared in normal prostate epithelial cells PrEC, prostate cancer cells PC-3M, and drug resistance cells PC-3M/ADM. PC-3M, PC-3M/ADM cell apoptosis and proliferation were detected by using flow cytometry under ADM treatment at IC50 concentration of PC-3M cells. PC-3M cells were cultured in vitro and divided into four groups, including microRNA-normal control (miR-NC), miR-203 mimic, small interfere NC (si-NC), and si-MEK1.
MiR-203 targeted and inhibited MEK1 expression. MiR-203 levels and cell apoptosis were significantly lower, while MEK1, p-ERK1/2, Bcl-2, and cell proliferation were significantly higher in PC-3M/ADM cells compared to the PC-3M cells. MiR-203 mimic and/or si-MEK1 transfection significantly reduced MEK1, p-ERK1/2, and Bcl-2 levels, attenuated cell proliferation, induced cell apoptosis, and decreased drug resistance.
MiR-203 elevation suppressed prostate cancer PC-3M cell proliferation, promoted apoptosis, and weakened ADM resistance through targeted inhibiting MEK1 expression to alleviate ERK/MAPK signaling pathway and Bcl-2 expression.
细胞外信号调节激酶（ERK）/丝裂原活化蛋白激酶（MAPK）信号通路广泛参与细胞增殖、凋亡和耐药。MAPK 激酶 1（MEK1）是 ERK 的上游蛋白激酶，可激活 ERK/MAPK 信号通路。研究发现微小 RNA 203（miR-203）下调与前列腺癌发病机制有关。生物信息学分析显示 miR-203 与 MEK1mRNA3'-UTR 之间存在互补的靶向关系。本研究通过影响 MEK1 表达，探讨 miR-203 调节前列腺癌细胞增殖、凋亡和 ADM 耐药的作用。
双荧光素酶报告基因实验证实 miR-203 与 MEK1 之间存在靶向关系。比较正常前列腺上皮细胞 PrEC、前列腺癌细胞 PC-3M 和耐药细胞 PC-3M/ADM 中 miR-203、MEK1、磷酸化 ERK1/2（p-ERK1/2）和 B 细胞淋巴瘤 2（Bcl-2）的表达。在 PC-3M 细胞的 IC50 浓度下，用 ADM 处理后通过流式细胞术检测 PC-3M/ADM 细胞的凋亡和增殖。体外培养 PC-3M 细胞，分为 miR-NC、miR-203 模拟物、si-NC 和 si-MEK1 四组。
miR-203 靶向并抑制 MEK1 的表达。与 PC-3M 细胞相比，PC-3M/ADM 细胞中的 miR-203 水平和细胞凋亡明显降低，而 MEK1、p-ERK1/2、Bcl-2 和细胞增殖明显升高。miR-203 模拟物和/或 si-MEK1 转染显著降低了 MEK1、p-ERK1/2 和 Bcl-2 的水平，减弱了细胞增殖，诱导了细胞凋亡，并降低了药物耐药性。
miR-203 上调通过靶向抑制 MEK1 的表达，抑制 ERK/MAPK 信号通路和 Bcl-2 的表达，抑制前列腺癌 PC-3M 细胞增殖，促进凋亡，减弱 ADM 耐药性。
