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Updated: Apr 29, 2026
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MiR-335 functions as a tumor suppressor in pancreatic cancer by targeting OCT4
Ling Gao1, Yijin Yang, Haiyan Xu
1Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, China.
Abstract:
Octamer-binding transcription factor 4 (OCT4) was closely related to pancreatic cancer progression, but its regulation in pancreatic cancer by microRNA (miRNA) is not fully clear. OCT4-positive and OCT4-negative pancreatic cells were isolated by flow cytometry, and it was found that OCT4-positive cells are enriched in transplanted pancreatic cancer cells compared with the primary ones and showed increasing proliferation and sphere formation. The data of miRNA array assay showed that miR-335 in OCT4-positive pancreatic cancer cells was lower than that in the negative ones. The results were confirmed in pancreatic cancer tissue and cell lines. Through expression analysis, it was found that miR-335 was underexpressed in OCT4(+) pancreatic cancer cells purified from primary tumors. Enforced expression of miR-335 in OCT4(+) pancreatic cancer cells inhibited clonogenic expansion and tumor development. miR-335 re-expression in OCT4(+) pancreatic cancer cells was blocked. Systemically delivered miR-335 inhibited pancreatic cancer metastasis and extended animal survival. Of significance, OCT4 was identified and validated as a direct and functional target of miR-335. Taken together, our results provide evidence that miR-335 might inhibit progression and stem cell properties of pancreatic cancer targeting OCT4.
Insights
MicroRNA-335 (miR-335) suppresses pancreatic cancer progression by targeting Octamer-binding transcription factor 4 (OCT4). Lower miR-335 levels correlate with increased tumor growth and metastasis, highlighting its therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- Octamer-binding transcription factor 4 (OCT4) is implicated in pancreatic cancer progression.
- The specific regulatory role of microRNAs (miRNAs) in OCT4-mediated pancreatic cancer is not fully understood.
Purpose of the Study:
- To investigate the relationship between miR-335 and OCT4 in pancreatic cancer.
- To determine if miR-335 can inhibit pancreatic cancer progression and stem cell properties.
Main Methods:
- Isolation of OCT4-positive and OCT4-negative pancreatic cancer cells using flow cytometry.
- miRNA array assay to compare miR-335 expression levels.
- Enforced expression of miR-335 in cancer cells and systemic delivery in vivo.
- Validation of OCT4 as a direct target of miR-335.
Main Results:
- OCT4-positive cells exhibited increased proliferation and sphere formation.
- miR-335 was significantly underexpressed in OCT4-positive pancreatic cancer cells and tissues.
- Enforced miR-335 expression inhibited tumor development, clonogenic expansion, and metastasis.
- OCT4 was confirmed as a direct functional target of miR-335.
Conclusions:
- miR-335 acts as a tumor suppressor in pancreatic cancer.
- Targeting OCT4 with miR-335 can inhibit pancreatic cancer progression and stem cell characteristics.
- miR-335 holds potential as a therapeutic agent for pancreatic cancer.
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