Molecular Genetics Department, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Molecular Genetics Department, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Gene. 2019 Sep 5;712:143958. doi: 10.1016/j.gene.2019.143958. Epub 2019 Jul 3.
The Wnt signaling pathway has been identified for its function in carcinogenesis and embryonic development. It is known to play a vital role in the initiation and development of colorectal cancer (CRC). Therefore, it is of great importance for CRC research to illuminate the mechanisms which regulate Wnt pathway activity. Here, we intended to examine the effect of hsa-miR-942 (miR-942) on the Wnt signaling activity, cell cycle progression, and its expression in CRC tissues. RT-qPCR results indicated that miR-942 is significantly upregulated in colorectal cancer. Then, overexpression of miR-942 promoted, whereas its inhibition decreased the Wnt signaling activity, detected by RT-qPCR and Top/Fop flash assay. Inhibition of Wnt signaling by using PNU-74654 or IWP-2 small molecules indicated that miR-942 applies its effect to the β-catenin degradation complex level. Then, RT-qPCR and dual luciferase assay showed that miR-942 upregulated Wnt signaling through direct targeting of APC, which is a tumor suppressor in Wnt signaling pathway. Furthermore, the western blotting analysis indicated that β.catenin, as a main member of Wnt signaling pathway is upregulated following the overexpression of miR-942. Finally, miR-942 overexpression resulted in cell cycle progression in SW480 cells. Taken together, our findings established an oncogenic role for miR-942 in CRC and indicated that this miRNA might be a crucial target for CRC therapy.
Wnt 信号通路因其在致癌和胚胎发育中的功能而被确定。它被认为在结直肠癌(CRC)的发生和发展中起着至关重要的作用。因此,阐明调节 Wnt 通路活性的机制对 CRC 研究非常重要。在这里,我们旨在研究 hsa-miR-942(miR-942)对 Wnt 信号活性、细胞周期进程及其在 CRC 组织中的表达的影响。RT-qPCR 结果表明 miR-942 在结直肠癌中显著上调。然后,miR-942 的过表达促进,而其抑制则降低了 Wnt 信号活性,通过 RT-qPCR 和 Top/Fop flash 测定检测。使用 PNU-74654 或 IWP-2 小分子抑制 Wnt 信号表明 miR-942 将其作用应用于 β-catenin 降解复合物水平。然后,RT-qPCR 和双荧光素酶报告基因实验表明,miR-942 通过直接靶向 APC 上调 Wnt 信号,APC 是 Wnt 信号通路中的肿瘤抑制因子。此外,Western blot 分析表明,β-catenin 作为 Wnt 信号通路的主要成员,在 miR-942 过表达后上调。最后,miR-942 的过表达导致 SW480 细胞的细胞周期进程。总之,我们的研究结果确立了 miR-942 在 CRC 中的致癌作用,并表明该 miRNA 可能是 CRC 治疗的关键靶点。