Department of General Surgery, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei 075061, P.R. China.
Mol Med Rep. 2020 Mar;21(3):1481-1490. doi: 10.3892/mmr.2020.10945. Epub 2020 Jan 16.
MicroRNA (miR)‑628‑5p serves as an antitumor gene in a variety of cancers; however, the role of miR‑628‑5p in colorectal cancer remains largely unclear. The purpose of this study was to investigate the role and mechanism of miR‑628‑5p in colorectal cancer. Reverse transcription‑quantitative PCR (RT‑qPCR), colony formation assays and flow cytometric analysis were used to determine the expression levels of miR‑628‑5p in colorectal cancer tissues and cell lines, and the proliferative ability of colorectal cancer cells. TargetScan version 7.2 and dual‑luciferase reporter assay were performed to predict and confirm miR‑628‑5p target genes. The expression levels of cyclin D1 (CCND1) and related genes were determined using RT‑qPCR or/and western blotting analysis. miR‑628‑5p mimics and CCND1 plasmids were used to overexpress miR‑628‑5p and CCND1; it was demonstrated that the expression levels of miR‑628‑5p were significantly downregulated in colorectal cancer tissues and cell lines. miR‑628‑5p mimic‑transfected cells inhibited the proliferation and induced apoptosis of HT‑29 cells. CCND1, a downstream effector of miR‑628‑5p, promoted the proliferation and suppressed apoptosis of HT‑29 cells, and the effects were reversed by miR‑628‑5p mimics. In conclusion, the present study suggested that colorectal cancer progression may be regulated through the miR‑628‑5p/CCND1 axis, and miR‑628‑5p could be used as a potential diagnostic and prognostic biomarker for colorectal cancer.
微小 RNA（miR）-628-5p 在多种癌症中作为抑癌基因发挥作用；然而，miR-628-5p 在结直肠癌中的作用在很大程度上仍不清楚。本研究旨在探讨 miR-628-5p 在结直肠癌中的作用和机制。采用逆转录定量 PCR（RT-qPCR）、集落形成实验和流式细胞术分析检测结直肠癌组织和细胞系中 miR-628-5p 的表达水平以及结直肠癌细胞的增殖能力。利用 TargetScan 版本 7.2 和双荧光素酶报告基因检测预测和验证 miR-628-5p 的靶基因。采用 RT-qPCR 或/和 Western blot 分析检测细胞周期蛋白 D1（CCND1）和相关基因的表达水平。使用 miR-628-5p 模拟物和 CCND1 质粒过表达 miR-628-5p 和 CCND1；结果表明，miR-628-5p 在结直肠癌组织和细胞系中的表达水平明显下调。miR-628-5p 模拟物转染的细胞抑制 HT-29 细胞的增殖并诱导其凋亡。CCND1 是 miR-628-5p 的下游效应因子，促进 HT-29 细胞的增殖并抑制其凋亡，而 miR-628-5p 模拟物则逆转了这些效应。综上所述，本研究表明结直肠癌的进展可能通过 miR-628-5p/CCND1 轴进行调节，miR-628-5p 可作为结直肠癌潜在的诊断和预后生物标志物。
