Department of Colorectal Surgery, Xinhua Affiliated Hospital of Dalian University, Dalian 116021, China.
Department of Endocrinology, The Second Hospital of Dalian Medical University, Dalian116023, China.
Eur J Pharmacol. 2018 Apr 15;825:75-84. doi: 10.1016/j.ejphar.2018.02.019. Epub 2018 Feb 15.
Colon cancer is one of the deadliest cancers worldwide; abnormal microRNA expression is common during colon cancer development. The aim of the present study was to elucidate the role played by miR-185 in this context. We used quantitative real-time PCR (qRT-PCR) to measure miR-185 expression levels in colon cancer cell lines. The effects of miR-185 on colon cancer cell proliferation and invasion were assessed using the MTT, colony-forming, wound-healing, and transwell assays. A luciferase activity assay was used to confirm the target of miR-185. Our data showed that miR-185 was significantly down-regulated in colon cancer cells and colonic cancer tissues compared with NCM460 normal colonic epithelial cells and adjacent normal tissues. A functional analysis revealed that ectopic expression of miR-185 significantly inhibited colon cancer cell proliferation, colony formation, migration, and invasion. In addition, western blot, qRT-PCR, and luciferase assays confirmed in colon cancer cells that Wnt1 was a downstream target of miR-185, in turn suppressing β-catenin-mediated signaling. In conclusion, we found that miR-185 inhibits colon cancer cell proliferation and invasion by targeting Wnt1, and that it serves as a tumor suppressor, indicating that the modulation of miR-185 levels may potentially be therapeutic in colon cancer patients.
结肠癌是全球最致命的癌症之一；在结肠癌的发展过程中，异常的 microRNA 表达是常见的。本研究的目的是阐明 miR-185 在这方面所起的作用。我们使用定量实时 PCR (qRT-PCR) 测量结肠癌细胞系中 miR-185 的表达水平。使用 MTT、集落形成、划痕愈合和 Transwell 测定评估 miR-185 对结肠癌细胞增殖和侵袭的影响。通过荧光素酶活性测定来证实 miR-185 的靶标。我们的数据表明，与 NCM460 正常结肠上皮细胞和相邻正常组织相比，miR-185 在结肠癌细胞和结肠癌细胞组织中明显下调。功能分析表明，miR-185 的异位表达显著抑制结肠癌细胞的增殖、集落形成、迁移和侵袭。此外，Western blot、qRT-PCR 和荧光素酶测定在结肠癌细胞中证实 Wnt1 是 miR-185 的下游靶标，进而抑制 β-连环蛋白介导的信号转导。总之，我们发现 miR-185 通过靶向 Wnt1 抑制结肠癌细胞的增殖和侵袭，并且作为肿瘤抑制因子，表明调节 miR-185 水平可能对结肠癌患者具有治疗潜力。
