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Targeting effect of microRNA on CD133 and its impact analysis on proliferation and invasion of glioma cells
1Department of Neurosurgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Abstract:
MiR-200b, a member of the microRNA-200 family, has been identified to be capable of suppressing glioma cell growth through targeting CREB1 or CD133. However, whether miR-200b affects the biological behavior (proliferation, invasion, and migration) of glioma cells is poorly understood. The aim of this study was to evaluate the effect of miR-200b on the biological behavior of glioma cells in vitro. MiRNA-200b mimics, miRNA-200b inhibitor, and mimic control were transfected into conventionally cultured glioma U251 cells, followed by measuring the expression of miR-200b and CD133 in transfected cells by RT-PCR; effect of miR-200b on CD133 mRNA 3'-UTR luciferase activity by luciferase reporter assay; proliferation activity of transfected U251 cells by MTT method; and changes in U251 cell invasion and migration by Transwell method after transfection. Compared to that in the miRNA-200b inhibitor, mimic control, and blank control groups, miRNA-200b expression was significantly increased and CD133 mRNA expression was significantly decreased in the mimic miRNA-200b group in a time-dependent manner (P < 0.05). Meanwhile, dual luciferase reporter assay showed that miR-200b could inhibit CD133 activity through binding to the 3'-UTR of CD133 mRNA (P < 0.05). Furthermore, the proliferation activity and invasion and migration abilities of U251 cells transfected with miRNA-200b mimic were significantly decreased (P < 0.05). In conclusion, overexpression of miR-200b inhibited the proliferation, invasion, and migration of glioma cells possibly through targeting CD133.
Insights
MicroRNA-200b (miR-200b) overexpression suppresses glioma cell proliferation, invasion, and migration. This occurs possibly by targeting CD133, a key factor in glioma progression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Glioma is a primary brain tumor with poor prognosis.
- MicroRNAs (miRNAs) are key regulators of gene expression.
- The role of miR-200b in glioma biological behavior is not fully understood.
Purpose of the Study:
- To investigate the effect of miR-200b on glioma cell proliferation, invasion, and migration.
- To explore the potential targeting of CD133 by miR-200b in glioma cells.
Main Methods:
- Transfection of glioma U251 cells with miR-200b mimics or inhibitors.
- Quantitative reverse transcription PCR (RT-PCR) to measure miR-200b and CD133 expression.
- Luciferase reporter assay to assess miR-200b targeting of CD133 mRNA.
- MTT assay for cell proliferation.
- Transwell assays for cell invasion and migration.
Main Results:
- miR-200b mimic transfection significantly increased miR-200b expression and decreased CD133 mRNA expression.
- miR-200b directly inhibited CD133 mRNA 3'-UTR luciferase activity.
- Overexpression of miR-200b significantly reduced glioma cell proliferation, invasion, and migration.
Conclusions:
- miR-200b plays an inhibitory role in glioma cell biological behavior.
- Targeting CD133 is a potential mechanism for miR-200b's anti-glioma effects.
- miR-200b represents a potential therapeutic target for glioma treatment.