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Characterization of Functionally Associated miRNAs in Glioblastoma and their Engineering into Artificial Clusters for Gene Therapy
Published on: October 4, 2019
miR-491 regulates glioma cells proliferation by targeting TRIM28 in vitro
Zengxin Qi1,2, Shengyong Cai1,2, Jiajun Cai1,2
1Department of Neurosurgery, Huashan Hospital, Fudan University, 12 Wulumuqi Middle Road, Shanghai, 200040, China.
Background:
MicroRNAs are significantly involved in tumorigenesis and progression of glioma. However, the critical part they play in glioma have not been fully elaborated. miR-491 and Tripartite motif containing 28 (TRIM28) are reported to aberrantly express in glioblastoma multiforme (GBM). Here, we detected miR-491 and TRIM28 expression and function in glioma cells.
Methods:
We analyzed miR-491 expressions in 20 primary human GBM tissues and 6 control brain tissues by qRT-PCR assays and searched for The Cancer Genome Atlas (TCGA) database. Then we predicted possible mRNA target of miR-491 by TargetScan/MicroRNA and confirmed it via luciferase reporter assays. Knock-down of miR-491 and transfection of pLenti-TRIM28 were performed in U251 and U87 cells. Proliferation ability was examined by MTT and clone formation assays.
Results:
miR-491 expression was obviously reduced in GBM cells and tissues. There was a positive correlation between the down-regulation of miR-491 and poor prognosis. Spearman's correlation analysis demonstrated that miR-491 expression was negatively correlated with TRIM28 protein level. Possible mRNA binding sites of miR-491 predicted by TargetScan/MicroRNA were proved by luciferase assays. Clone formation and MTT assays indicated that up-regulation of miR-491 inhibited the proliferation of glioma cells.
Conclusions:
miR-491 regulates glioma cells proliferation in vitro by targeting TRIM28.
Insights
MicroRNA 491 (miR-491) is downregulated in glioma and inhibits tumor cell proliferation by targeting Tripartite motif containing 28 (TRIM28). This finding offers new insights into glioma progression and potential therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- MicroRNAs play a crucial role in tumorigenesis and glioma progression.
- Aberrant expression of miR-491 and Tripartite motif containing 28 (TRIM28) is observed in glioblastoma multiforme (GBM).
- The precise function of miR-491 in glioma requires further elucidation.
Purpose of the Study:
- To investigate the expression and functional role of miR-491 in glioma.
- To determine the relationship between miR-491 and TRIM28 in glioma cells.
Main Methods:
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays were used to analyze miR-491 expression in GBM tissues and control brain tissues.
- The Cancer Genome Atlas (TCGA) database was searched for relevant expression data.
- Bioinformatic tools (TargetScan/MicroRNA) predicted miR-491 targets, which were validated using luciferase reporter assays.
- Functional assays, including MTT and clone formation assays, were performed after miR-491 knockdown and TRIM28 transfection in U251 and U87 glioma cell lines.
Main Results:
- miR-491 expression was significantly reduced in GBM tissues and cells compared to controls.
- Down-regulation of miR-491 correlated positively with poor prognosis in glioma patients.
- A negative correlation was observed between miR-491 expression and TRIM28 protein levels.
- Upregulation of miR-491 suppressed glioma cell proliferation in vitro.
Conclusions:
- miR-491 acts as a tumor suppressor in glioma by targeting TRIM28.
- miR-491 regulates glioma cell proliferation in vitro.
- These findings highlight the potential of targeting miR-491 for glioma therapy.
