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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
MALAT1 Expression Is Deregulated in miR-34a Knockout Cell Lines
Andrea Corsi1, Tonia De Simone1, Angela Valentino1
1Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Strada le Grazie 8, 37134 Verona, Italy.
Abstract:
Background/Objectives: Non-coding microRNA-34a (miR-34a) regulates the expression of key factors involved in several cellular processes, such as differentiation, apoptosis, proliferation, cell cycle, and senescence. Deregulation of the expression of these factors is implicated in the onset and progression of several human diseases, including cancer, neurodegenerative disorders, and pathologies associated with viral infections and inflammation. Despite numerous studies, the molecular mechanisms regulated by miR-34a remain to be fully understood. The present study aimed to generate miR-34a knockout cell lines to identify novel genes potentially regulated by its expression. Methods: We employed the CRISPR-Cas9 gene editing system to knock out the hsa-miR-34a gene in HeLa and 293T cell lines, two widely used models for studying molecular and cellular mechanisms. We compared proliferation rates and gene expression profiles via RNA-seq and qPCR analyses between the wild-type and miR-34a KO cell lines. Results: Knockout of miR-34a resulted in a decreased proliferation rate in both cell lines. Noteworthy, the ablation of miR-34a resulted in increased expression of the long non-coding RNA MALAT1. Additionally, miR-34a-5p silencing in the A375 melanoma cell line led to MALAT1 overexpression. Conclusions: Our findings support the role of the miR-34a/MALAT1 axis in regulating proliferation processes.
Insights
MicroRNA-34a (miR-34a) knockout cells showed reduced proliferation and increased MALAT1 expression, revealing a novel miR-34a/MALAT1 axis that regulates cell growth.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- MicroRNA-34a (miR-34a) is a non-coding microRNA crucial for regulating cellular processes like differentiation, apoptosis, and proliferation.
- Dysregulation of miR-34a is linked to various diseases, including cancer and inflammatory conditions.
- The precise molecular mechanisms governed by miR-34a are not fully elucidated.
Purpose of the Study:
- To generate miR-34a knockout cell lines to identify novel genes regulated by miR-34a.
- To investigate the functional impact of miR-34a ablation on cellular processes.
Main Methods:
- Utilized CRISPR-Cas9 gene editing to create miR-34a knockout HeLa and 293T cell lines.
- Assessed proliferation rates and analyzed gene expression profiles using RNA-seq and qPCR.
- Validated findings by examining MALAT1 expression following miR-34a-5p silencing in melanoma cells.
Main Results:
- miR-34a knockout led to a significant decrease in cell proliferation in both cell lines.
- Ablation of miR-34a resulted in the upregulation of the long non-coding RNA MALAT1.
- miR-34a-5p silencing in melanoma cells also caused MALAT1 overexpression.
Conclusions:
- The study identified a novel regulatory axis involving miR-34a and MALAT1.
- This miR-34a/MALAT1 axis plays a role in the regulation of cellular proliferation processes.

