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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
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MicroRNA‑628‑5p inhibits cell proliferation and induces apoptosis in colorectal cancer through downregulating CCND1

Fei Guo1, Jun Xue1

  • 1Department of General Surgery, The First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei 075061, P.R. China.

Molecular Medicine Reports
|February 5, 2020
PubMed

Insights

MicroRNA-628-5p is downregulated in colorectal cancer, inhibiting proliferation and inducing apoptosis. Its axis with cyclin D1 (CCND1) suggests miR-628-5p as a potential biomarker for colorectal cancer.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • MicroRNA (miR)-628-5p exhibits antitumor properties in various cancers.
  • The specific function of miR-628-5p in colorectal cancer (CRC) is not well-defined.
  • Understanding miR-628-5p's role is crucial for developing novel CRC therapeutic strategies.

Purpose of the Study:

  • To elucidate the role and underlying mechanism of miR-628-5p in colorectal cancer.
  • To investigate miR-628-5p's impact on CRC cell proliferation and apoptosis.
  • To identify potential therapeutic targets within the miR-628-5p pathway.

Main Methods:

  • Quantitative PCR (RT-qPCR) and western blotting to assess gene expression.
  • Colony formation assays and flow cytometry for proliferation and apoptosis analysis.
  • Bioinformatic prediction (TargetScan) and dual-luciferase reporter assays to identify miR-628-5p targets.

Main Results:

  • miR-628-5p expression was significantly downregulated in colorectal cancer tissues and cell lines.
  • Overexpression of miR-628-5p inhibited proliferation and induced apoptosis in HT-29 cells.
  • Cyclin D1 (CCND1) was identified as a direct target of miR-628-5p, promoting proliferation and suppressing apoptosis; these effects were reversed by miR-628-5p mimics.

Conclusions:

  • Colorectal cancer progression is potentially regulated by the miR-628-5p/CCND1 axis.
  • miR-628-5p demonstrates potential as a diagnostic and prognostic biomarker for colorectal cancer.
  • Targeting the miR-628-5p/CCND1 pathway may offer a novel therapeutic approach for colorectal cancer.
Keywords:
colorectal cancermicrorna-628-5pproliferationapoptosiscyclin d1

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