Predicted miRNAs suppressed cell proliferation and migration via FAK/VASP axis; Systems biology approach
Asghar Mohammadi, Ghasem Ghasempour, Abbas Sahebghadam Lotfi, Mohammad Najafi
Abstract
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. 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. 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). 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.