Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, KY, 40292, USA.
James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, USA.
BMC Cancer. 2018 Apr 13;18(1):421. doi: 10.1186/s12885-018-4258-0.
Dysregulation of microRNA (miRNA) expression is associated with hallmarks of aggressive tumor phenotypes, e.g., enhanced cell growth, proliferation, invasion, and anchorage independent growth in prostate cancer (PCa).
Serum-based miRNA profiling involved 15 men diagnosed with non-metastatic (stage I, III) and metastatic (stage IV) PCa and five age-matched disease-free men using miRNA arrays with select targets confirmed by quantitative real-time PCR (qRT-PCR). The effect of miR-186-5p inhibition or ectopic expression on cellular behavior of PCa cells (i.e., PC-3, MDA-PCa-2b, and LNCaP) involved the use bromodeoxyuridine (BrdU) incorporation, invasion, and colony formation assays. Assessment of the impact of miR-186-5p inhibition or overexpression on selected targets entailed microarray analysis, qRT-PCR, and/or western blots. Statistical evaluation used the modified t-test and ANOVA analysis.
MiR-186-5p was upregulated in serum from PCa patients and metastatic PCa cell lines (i.e., PC-3, MDA-PCa-2b, LNCaP) compared to serum from disease-free individuals or a normal prostate epithelial cell line (RWPE1), respectively. Inhibition of miR-186-5p reduced cell proliferation, invasion, and anchorage-independent growth of PC-3 and/or MDA-PCa-2b PCa cells. AKAP12, a tumor suppressor target of miR-186-5p, was upregulated in PC-3 and MDA-PCa-2b cells transfected with a miR-186-5p inhibitor. Conversely, ectopic miR-186-5p expression in HEK 293 T cells decreased AKAP12 expression by 30%. Both pAKT and β-catenin levels were down-regulated in miR-186-5p inhibited PCa cells.
Our findings suggest miR-186-5p plays an oncogenic role in PCa. Inhibition of miR-186-5p reduced PCa cell proliferation and invasion as well as increased AKAP12 expression. Future studies should explore whether miR-186-5p may serve as a candidate prognostic indicator and a therapeutic target for the treatment of aggressive prostate cancer.
微小 RNA（miRNA）表达失调与侵袭性肿瘤表型的标志有关，例如前列腺癌（PCa）中增强的细胞生长、增殖、侵袭和锚定非依赖性生长。
使用 miRNA 阵列对 15 名诊断为非转移性（I 期、III 期）和转移性（IV 期）PCa 以及 5 名年龄匹配的无疾病男性进行基于血清的 miRNA 分析，并使用定量实时 PCR（qRT-PCR）确认了选定的靶标。miR-186-5p 抑制或异位表达对 PCa 细胞（即 PC-3、MDA-PCa-2b 和 LNCaP）的细胞行为的影响涉及溴脱氧尿苷（BrdU）掺入、侵袭和集落形成测定。评估 miR-186-5p 抑制或过表达对选定靶标的影响需要进行微阵列分析、qRT-PCR 和/或 Western blot。使用修改后的 t 检验和 ANOVA 分析进行统计评估。
与无疾病个体或正常前列腺上皮细胞系（RWPE1）的血清相比，miR-186-5p 在 PCa 患者和转移性 PCa 细胞系（即 PC-3、MDA-PCa-2b、LNCaP）的血清中上调。miR-186-5p 抑制降低了 PC-3 和/或 MDA-PCa-2b PCa 细胞的增殖、侵袭和锚定非依赖性生长。AKAP12 是 miR-186-5p 的肿瘤抑制靶标，在转染 miR-186-5p 抑制剂的 PC-3 和 MDA-PCa-2b 细胞中上调。相反，在 HEK 293 T 细胞中外源表达 miR-186-5p 使 AKAP12 表达降低了 30%。miR-186-5p 抑制的 PCa 细胞中 pAKT 和 β-连环蛋白水平下调。
我们的研究结果表明 miR-186-5p 在 PCa 中发挥致癌作用。miR-186-5p 抑制降低了 PCa 细胞的增殖和侵袭，并增加了 AKAP12 的表达。未来的研究应探讨 miR-186-5p 是否可作为侵袭性前列腺癌的候选预后指标和治疗靶标。
