The Silencing Effect of Mir-30A on Itga4 Gene Expression in Vitro: An Approach for Gene Therapy Publisher



Darzi L¹ ; Boshtam M² ; Shariati L² ; Kouhpayeh S³ ; Gheibi A⁴ ; Mirian M⁵ ; Rahimmanesh I¹ ; Khanahmad H^{1, 6} ; Tabatabaiefar MA^{1, 6} Show Affiliations
Source: Research in Pharmaceutical Sciences Published:2017

Abstract

Integrins are adhesion molecules which play crucial roles in cell-cell and cell-extracellular matrix interactions. Very late antigen-4 or 4β1 and lymphocyte Peyer's patch adhesion molecule-1 or 4β7, are key factors in the invasion of tumor cells and metastasis. Based on the previous reports, integrin 4 (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.