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Updated: Dec 20, 2025
Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
Hsa-miR-3658 down-regulates OCT4 gene expression followed by suppressing SW480 cell proliferation and migration
Fahimeh Hosseini1, Bahram M Soltani1, Hossein Baharvand2,3
1Genetics Department, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Abstract:
The pluripotency factor, OCT4 gene is a stemness marker that is involved in the tumorigenicity of different cancer types and knowing about molecular mechanisms of its regulation is crucially important. To date, a few microRNAs (miRNAs) are known to be regulators of OCT4 gene expression. Looking for the novel miRNAs which are capable of regulating OCT4 gene expression, our bioinformatics analysis introduced hsa-miR-3658 (miR-3658) as a bona fide candidate. Then, RT-qPCR results indicated that miR-3658 expression is decreased in colorectal cancer (CRC) tumor tissues, compared with normal pairs. Furthermore, RT-qPCR and western blot analysis showed that the OCT4 gene has been down-regulated following the miR-3658 overexpression. Consistently, dual-luciferase assay supported the direct interaction of miR-3658 with the 3'-UTR sequence of OCT4 gene. Unlike in HCT116 cells, overexpression of miR-3658 in SW480 cells brought about growth inhibition, cell cycle arrest and reduced cell migration, detected by flow cytometry, and scratch test assay. Overall, these findings demonstrated that miR-3658 as a tumor suppressor miRNA exerts its effect against OCT4 gene expression, and it has the potential of being used as a prognostic marker and therapeutic target against colorectal cancer.
Insights
A novel microRNA, miR-3658, acts as a tumor suppressor by down-regulating OCT4 expression in colorectal cancer. This finding highlights miR-3658 as a potential prognostic marker and therapeutic target for CRC.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- OCT4 is a pluripotency factor and stemness marker implicated in cancer tumorigenicity.
- Understanding OCT4 gene regulation is crucial for cancer research.
- MicroRNAs (miRNAs) are known regulators of gene expression, including OCT4.
Purpose of the Study:
- To identify novel miRNAs regulating OCT4 gene expression.
- To investigate the role of hsa-miR-3658 (miR-3658) in colorectal cancer (CRC).
Main Methods:
- Bioinformatics analysis to identify candidate miRNAs.
- RT-qPCR to assess miR-3658 and OCT4 expression.
- Western blot to confirm OCT4 protein levels.
- Dual-luciferase assay to validate direct interaction.
- Cell-based assays (flow cytometry, scratch test) to evaluate functional effects.
Main Results:
- Bioinformatics identified miR-3658 as a potential regulator of OCT4.
- miR-3658 expression was decreased in CRC tissues.
- miR-3658 overexpression led to OCT4 down-regulation.
- Direct interaction between miR-3658 and OCT4 3'-UTR was confirmed.
- miR-3658 overexpression inhibited cell growth, induced cell cycle arrest, and reduced migration in SW480 cells.
Conclusions:
- miR-3658 functions as a tumor suppressor miRNA in colorectal cancer.
- miR-3658 exerts its tumor-suppressive effects by targeting OCT4.
- miR-3658 holds potential as a prognostic biomarker and therapeutic target for CRC.
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