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Published on: January 22, 2021
miR-194 Inhibits the Proliferation of SW620 Colon Cancer Stem Cells Through Downregulation of SSH2 Expression
Bo Sun1, Yan-Tian Fang1, Dan-Juan Jin2
1Department of Gastric Cancer, Fudan University Shanghai Cancer Center, Shanghai, People's Republic of China.
Purpose:
Colorectal cancer (CRC) stem cells are tumorigenic, capable of self-renewal, and resistant to therapy. Although the expression pattern and functions of micro RNA (miR)-194 in CRC cells have been widely investigated, little is known about its role in CRC stem cells. Therefore, the aim of this study was to investigate the potential role of miR-194 in CRC stem cells.
Materials And Methods:
CRC stem cells were isolated from the SW620 colon cancer cell line using microbeads. The expression levels of miR-194 and slingshot 2 (SSH2) in CRC stem cells were detected by RT-PCR and Western blot. A luciferase reporter assay was performed to confirm that miR-194 directly targets SSH2. Proliferation of CRC stem cells was examined by colony formation and MTT assays. Apoptosis in CRC stem cells was detected by cell cycle and apoptosis assays. The role of miR-194 in tumor growth was determined in vivo.
Results:
Cells positive for CD44 and CD133 accounted for approximately 88.7% of the isolated population after microbead isolation. We reveal for the first time that miR-194 expression is decreased in CRC stem cells. Specifically, miR-194 is involved in inhibiting the proliferation of CRC stem cells and promoting CRC stem cell apoptosis by directly targeting SSH2. Furthermore, overexpression of miR-194 resulted in blocking the G1/S transition, the induction of cellular apoptotic process, thereby suppressing the malignant behaviors of CRC stem cells.
Conclusion:
This study represents a novel characterization of miR-194 function in CRC stem cells, which may aid in the development of promising therapeutic strategies targeting CRC.
Insights
MicroRNA-194 (miR-194) is decreased in colorectal cancer (CRC) stem cells. This study shows miR-194 inhibits CRC stem cell proliferation and promotes apoptosis by targeting SSH2, offering potential therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Colorectal cancer (CRC) stem cells possess tumorigenic, self-renewal, and therapy-resistant properties.
- While microRNA-194 (miR-194) is studied in CRC cells, its role in CRC stem cells remains largely unknown.
Purpose of the Study:
- To investigate the function of miR-194 in colorectal cancer stem cells.
Main Methods:
- Isolation of CRC stem cells from the SW620 cell line.
- Quantification of miR-194 and SSH2 expression using RT-PCR and Western blot.
- Luciferase reporter assay to confirm direct targeting of SSH2 by miR-194.
- Assessment of cell proliferation, apoptosis, and in vivo tumor growth.
Main Results:
- miR-194 expression is significantly decreased in colorectal cancer stem cells.
- miR-194 directly targets SSH2, inhibiting stem cell proliferation and inducing apoptosis.
- Overexpression of miR-194 blocks G1/S transition, promotes apoptosis, and suppresses malignant behaviors.
Conclusions:
- This study elucidates a novel function of miR-194 in colorectal cancer stem cells.
- miR-194's role in suppressing malignant behaviors suggests its potential as a therapeutic target for CRC.
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