前列腺癌miRNA表达谱及miR-96在氧化应激信号通路中的作用

Expression Profile of MicroRNAs in Prostate Carcinoma and Role of miR-96 in Regulation of Oxidative Stress

【目的】检测miRNA在前列腺癌与正常前列腺组织的表达差异,并探讨表达异常的miRNA在前列腺癌发病中的作用。【方法】前列腺癌和正常前列腺组织样品(100 mg),Trizol法提取RNA,应用聚合酶链反应(PCR)芯片检测miRNA的表达;同时以250μmol/L的过氧化氢(H2O2)刺激前列腺癌PC-3细胞4 h,继续正常培养12 h。检测不同时间点细胞内活性氧(ROS)水平,以实时荧光定量PCR(qRT-PCR)检测miR-96在各组细胞中的表达。【结果】与正常前列腺组织相比,前列腺癌组织中miR-144、miR-216a上调5.9倍,miR-96上调30.4倍,miR-488、miR-873下调到4.9%,差异有统计学意义(P<0.05)。PC-3细胞内ROS为RWPE-1细胞的5.2倍(P<0.05),miR-96在PC-3细胞中的表达为RWPE-1细胞的15.4倍(P<0.05)。H2O2刺激PC-3细胞1、4 h后,胞内ROS为对照组的4.3、6.4倍(P均<0.05),miR-96表达水平为对照组的10.2、18.9倍(P均<0.05);10、16 h组胞内ROS为对照组的2.5倍、1.2倍,miR-96表达水平为对照组的2.7、1.9倍(P均>0.05)。【结论】在前列腺癌组织中发现了若干有表达差异的miRNA;miR-96参与前列腺癌细胞的氧化应激信号途径,可能是防治前列腺癌的重要分子靶点。

[ Objective ] To investigate the expression profile of miRNAs in the prostate cancer and benign prostatic tissues, and to evaluate the role of differential miRNAs in the regulation of oxidative signaling of prostate cancer. [Methods] The total RNA was extracted from prostate cancer tissues and adjacent non-tumorous tissues （ 100 mg each） and the expression of miRNAs were detected by miRNA PCR Array （Denmark, Exiqon）. Prostate cancer cell line PC-3 was incubated with hydrogen peroxide （H202, 250 p^mol/L） for 4 hours, and then was cultured in normal condition for 12 hours. The intracellular reactive oxygen species （ROS） and the expression of miR-96 were detected at indicated time points. [ Results ] Compared to the normal prostate tissues, miR-144, and miR- 216a were up-regulated 5.9 times, and miR-96 was up-regulated 30.4 times in the prostate cancer tissues, while miR-488 and miR- 873 were down-regulated to 4.9%（P〈0.05）. The intracellular ROS of PC-3 cells was as 5.2 times as to that in the RWPE-I cells （P 〈 0.05）, and the expression of miR-96 in PC-3 was 15.4 times compared to that in the RWPE-1 cells （P 〈 0.05）. In the 1 h and 4 h group, H202 induced 4.3 and 6.4 times of ROS compared to that in the control （P 〈 0.05）, with 10.2 and 18.9 times of miR-96 expression compared to that in the control （P 〈 0.05 ）. In the l0 h and 16 h group, the intracellular ROS remained 2.5 and 1.2 times compared to the control （P 〉 0.05）, with 2.7 and 1.9 times of miR-96 expression compared to the control （P 〉 0.05）. [Conclusion] This study had identified several different expression miRNAs in prostate cancer, miR-96 plays an important role in the oxidative signaling of prostate cancer. It may be developed as an important molecular target for the treatment of prostate cancer.