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

Leila Darzi, Maryam Boshtam, Laleh Shariati, Shirin Kouhpayeh, Azam Gheibi, Mina Mirian, Ilnaz Rahimmanesh, Hossein Khanahmad, Mohammad Amin Tabatabaiefar

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.


Keywords


Cancer; ITGA4; Metastasis; miR-30a: Non-coding RNA

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References


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