The silencing effect of miR-30a on ITGA4 gene expression in vitro: an approach for gene therapy

Abstract

Integrins are adhesion molecules which play crucial roles in cell-cell and cell-extracellular matrix interactions. Very late antigen-4 or α4β1 and lymphocyte Peyer’s patch adhesion molecule-1 or α4β7, are key factors in the invasion of tumor cells and metastasis. Based on the previous reports, integrin α4 (ITGA4) is overexpressed in some immune disorders and cancers. Thus, inhibition of ITGA4 could be a therapeutic strategy. In the present study, miR-30a was selected in order to suppress ITGA4 expression. The ITGA4 3' UTR was amplified, cloned in the Z2827-M67-(ITGA4) plasmid and named as Z2827-M67/3'UTR. HeLa cells were divided into five groups; (1) untreated without any transfection, (2) mock with Z2827-M67/3'UTR transfection and X-tremeGENE reagent, (3) negative control with Z2827-M67/3'UTR transfection alone, (4) test with miR-30a mimic and Z2827-M67/3'UTR transfection and (5) scramble with miR-30a scramble and Z2827-M67/3'UTR transfection. The MTT assay was performed to evaluate cell survival and cytotoxicity in each group. Real-time RT-PCR was applied for the ITGA4 expression analysis. The findings of this study showed that miR-30a downregulated ITGA4 expression and had no effect on the cell survival. Due to the silencing effect of miR-30a on the ITGA4 gene expression, this agent could be considered as a potential tool for cancer and immune disorders therapy.

Construction and characterization of human embryonic kidney-(HEK)-293T cell overexpressing truncated α4 integrin

“…Integrins exist as two non-covalently bound α and β subunits, which pair to form heterodimers( 4 5 ). Integrins can bind to extracellular matrix (ECM) glycoproteins including collagens, fibronectins, laminins, and cellular receptors such as VCAM-1 and the intercellular cell adhesion molecule (ICAM) family( 5 6 ).…”

Screening of differentially expressed immune-related genes from spleen of broilers fed with probiotic Bacillus cereus PAS38 based on suppression subtractive hybridization

“…They are important cell surface receptors. They mediate adhesion of cells to extracellular matrices and adhesion of cell to cell, participate in cell proliferation, differentiation, adhesion, migration and other processes, and play an important role in the growth and development, immune response and other physiological processes of the body [54, 55]. Kim et al [56] found that the down-regulation of ITGA4 in duodenum of chickens infected with Eimeria .…”

Chimeric antigen receptor-T cells immunotherapy for targeting breast cancer

“…Some important advances in genomic techniques have led to recognize possible biomarkers involved in breast cancer development, progression, and invasion ( 1 2 ). Currently, personalized approaches towards the prediction and management of different subtypes of breast cancer have been developed based on omics data ( 3 4 5 ). Despite significant progress in treatment strategies, certain breast tumors remain resistant to the current treatments due to their heterogeneous nature.…”