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Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells
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MiR-17-5p and MKL-1 modulate stem cell characteristics of gastric cancer cells

Zhou-Tong Dai1, Yuan Xiang2, Yuan-Yuan Duan1

  • 1Institute of Biology and Medicine, College of Life and Health Sciences, Wuhan University of Science and Technology, Hubei, 430081, P.R. China.

International Journal of Biological Sciences
|July 9, 2021
PubMed

Insights

This study reveals microRNA miR-17 and MKL-1 promote gastric cancer stem cell properties. Targeting these may offer new therapeutic strategies for gastric cancer.

Area of Science:

  • Molecular Oncology
  • Cancer Stem Cell Biology
  • Gastric Cancer Research

Background:

  • Targeting cancer stem cells (CSCs) holds significant therapeutic promise for cancer treatment.
  • The roles of microRNA (miRNA) and epithelial-mesenchymal transition (EMT) in regulating CSC characteristics are well-established.
  • The specific involvement of miR-17 and MKL-1 in gastric CSCs remained unexplored.

Purpose of the Study:

  • To investigate the mechanism of miR-17 and MKL-1 in gastric cancer stem cells.
  • To explore the potential of miR-17 and MKL-1 as therapeutic targets in gastric cancer.

Main Methods:

  • Utilized RT-PCR, Western Blot, flow cytometry, and immunofluorescence for cellular analysis.
  • Employed sphere formation assays, xenograft tumor models, and drug resistance tests in vitro and vivo.
  • Verified MKL-1 targets (CD44, EpCAM, miR-17-5p promoter) using luciferase assays and ChIP.

Main Results:

  • miR-17-5p and MKL-1 were confirmed to promote stem cell-like properties in gastric cancer.
  • MKL-1 was found to target the promoters of CD44, EpCAM, and miR-17-5p.
  • TCGA database analysis showed increased miR-17-5p and MKL-1 in gastric cancer, with higher MKL-1 correlating with reduced survival.

Conclusions:

  • Identified a novel regulatory pathway involving miR-17 and MKL-1 in gastric CSCs.
  • These findings provide new insights into potential early diagnosis and molecular therapies for gastric cancer.
  • MKL-1 promotes gastric cancer progression by targeting miR-17, CD44, and EpCAM.
Keywords:
MKL-1bioinformatics.gastric cancermiR-17-5pstem cells

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