The Second Xiangya Hospital, Central South University, Changsha, China.
Clinical Medical College, Yangzhou University, Yangzhou, China.
Acta Biochim Biophys Sin (Shanghai). 2019 Jul 10;51(7):661-668. doi: 10.1093/abbs/gmz047.
The functional role of microRNA-23a in tumorigenesis has been investigated; however, the exact mechanism of microRNA-23a (miR-23a) in colorectal cancer development has not been fully explored. In the present study, we aimed to investigate the molecular functional role of miR-23a in colorectal carcinogenesis. Quantitative real-time polymerase chain reaction was conducted to investigate the expression level of miR-23a in tissue samples and cell lines (HCT116 and SW480). CCK-8, colony formation and Transwell assay were used to explore the role of miR-23a in cell proliferation and migration. Dual luciferase reporter assay was used to identify the direct binding of miR-23a with its target, MARK1. Western blot analysis was used to analyze the expression level of MARK1, as well as a confirmed miR-23a target gene, MTSS1, in miR-23a-mimic and miR-23a-inhibit groups. Rescue experiments were conducted by overexpression of MARK1 in miR-23a-mimic-transfected cell lines. The results showed that miR-23a was highly expressed in colorectal cancer tissue and cell lines. MiR-23a could promote proliferation and migration of colorectal cancer cell lines. MARK1 was a direct target of miR-23a and the expression level of MARK1 was down-regulated in miR-23a-mimic-transfected cell lines but up-regulated in miR-23a-inhibit-transfected cells. Overexpression of MARK1 could partly reverse the cancer-promoting function of miR-23a. Our results suggested that miR-23a promotes colorectal cancer cell proliferation and migration by mediating the expression of MARK1. MiR-23a may be a potential therapeutic target for colorectal cancer treatment.
microRNA-23a 在肿瘤发生中的功能作用已经得到了研究；然而，microRNA-23a（miR-23a）在结直肠癌发展中的确切机制尚未完全探索。在本研究中，我们旨在研究 miR-23a 在结直肠癌细胞发生中的分子功能作用。采用实时定量聚合酶链反应检测组织样本和细胞系（HCT116 和 SW480）中 miR-23a 的表达水平。CCK-8、集落形成和 Transwell 测定用于探索 miR-23a 在细胞增殖和迁移中的作用。双荧光素酶报告基因检测用于鉴定 miR-23a 与其靶基因 MARK1 之间的直接结合。Western blot 分析用于分析 miR-23a 模拟物和 miR-23a 抑制剂组中 MARK1 以及已确认的 miR-23a 靶基因 MTSS1 的表达水平。通过在 miR-23a 模拟物转染的细胞系中转染 MARK1 的过表达进行挽救实验。结果表明，miR-23a 在结直肠癌组织和细胞系中高表达。miR-23a 可促进结直肠癌细胞系的增殖和迁移。MARK1 是 miR-23a 的直接靶基因，在 miR-23a 模拟物转染的细胞系中 MARK1 的表达水平下调，而在 miR-23a 抑制剂转染的细胞中上调。MARK1 的过表达可部分逆转 miR-23a 的促癌功能。我们的结果表明，miR-23a 通过调节 MARK1 的表达促进结直肠癌细胞的增殖和迁移。miR-23a 可能是结直肠癌治疗的潜在治疗靶点。
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