Predicted miRNAs suppressed cell proliferation and migration via FAK/VASP axis; Systems biology approach

Abstract

Introduction: Many studies have reported that some genes in cellular proliferation and migration pathways relate to CRC pathogenesis. The aim of this study was to predict bioinformatically important gene-related miRNAs and investigate experimentally the changes of protein values and gene expression levels, and evaluate the cell proliferation and migration in miRNA/PEI-transfected HT-29 CRC cells. Methods: The gene and miRNA networks were predicted using bioinformatics data and tools. The gene-related miRNAs were transfected using PEI particles into HT-29 CRC cells. The gene expression levels and protein values were measured by RT-qPCR and western techniques, respectively. The cell viability was evaluated by MTT technique. The cell migration was measured by scratch assay. The cell miR/PEI transfection was confirmed by fluorescence microscope, flow cytometry, and electronic microscope. Results: The FAK, VASP, miR-520a and miR-20a were predicted on the gene-miRNA networks. The FAK gene expression levels (P = 0.014) and protein values (P = 0.0001) decreased significantly in miR-520a-3p/PEI-transfected cells. Furthermore, the VASP gene expression levels (P = 0.01) and protein values (P = 0.0001) decreased significantly by miR-20a-5p. The data also showed that the cell migration reduced significantly in the transfected cells with miR-20a-5p (P < 0.0001) and miR-520a-3p (P < 0.0001). Conclusion: The results confirmed the roles of FAK and VASP genes in the cellular proliferation and migration and showed that miR-20a-5p and miR-520a-3p suppress these events in HT-29 cells.

Keywords

• CRC
• FAK
• VASP
• miR-520a-3p
• miR-20a-5p
• Migration