Micro-RNA-186-5p 抑制可减弱转移性前列腺癌细胞的增殖、锚定非依赖性生长和侵袭。

BACKGROUND: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

背景：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。

方法：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。

结果：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。

结论：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。