Effect of miR-۵۷۹ overexpression on the BAX and CDKN۱A genes in the Glioblastoma cell line

Aims: Glioblastoma multiforme is a type of brain cancers that do not respond well to treatment. The poor prognosis of this disease is due to the presence of radiation resistance and chemotherapy. The purpose of the present study was to produce miR-۵۷۹ precursor carriers and investigate the effect of increased expression of miR-۵۷۹ on the expression of BAX and CDKN۱A genes in the glioblastoma cell line. Materials and Methods: In this experimental study, in order to produce recombinant lentiviral vectors, a gene containing the miR-۵۷۹ precursor sequence was cloned into the plasmid. The recombinant structure was transmitted to the cells of HEK۲۹۳T with and pMD۲ plasmids. Viral particles were concentrated using Ultra Centrifuge. Viral titration was calculated by flow cytometry. The viral particles produced were transferred to the A-۱۷۲ cell line. Finally, by using Real-Time PCR, changes in expression levels of miR-۵۷۹ and BAX and CDKN۱A genes were investigated. Findings: The presence of miR-۵۷۹ gene precursor in the plasmid was confirmed by colony PCR and sequencing methods. The study showed that the level of miR-۵۷۹ expression in infected cells with the recombinant virus was found to be up-regulated compared to the control group. miR-۵۷۹ increased the BAX gene expression by three times. But, there was no significant change in the expression of CDKN۱A gene expression. Conclusion: Increased expression of miR-۵۷۹ in the A-۱۷۲ cell line could increase the expression of BAX gene. However, the CDKN۱A gene expression does not change significantly.

کلیدواژه ها:

miR-۵۷۹ ، Glioblastoma ، CDKN۱A ، BAX ، miR-۵۷۹ ، گلیوبلاستوما ، BAX ، CDKN۱A