Volume 22, Issue 1 (2018)
mjms 2018, 22(1): 27-34 |
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Kalhori M, Irani S, Soleimani M, Arefian E, Kouhkan F. Effect of miR-579 overexpression on the BAX and CDKN1A genes in the Glioblastoma cell line. mjms 2018; 22 (1) :27-34
URL: http://mjms.modares.ac.ir/article-30-16349-en.html
URL: http://mjms.modares.ac.ir/article-30-16349-en.html
1- Biology Department, Science & Research Branch, Islamic Azad University, Tehran, Iran
2- Hematology Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran ,soleim_m@modares.ac.ir
3- Microbiology Department, Biology Faculty, College of Science, University of Tehran, Tehran, Iran
4- Molecular Biology & Genetic Engineering Department, Stem Cell Technology Research Center, Tehran, Iran
2- Hematology Department, Medical Sciences Faculty, Tarbiat Modares University, Tehran, Iran ,
3- Microbiology Department, Biology Faculty, College of Science, University of Tehran, Tehran, Iran
4- Molecular Biology & Genetic Engineering Department, Stem Cell Technology Research Center, Tehran, Iran
Abstract: (8713 Views)
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-579 precursor carriers and investigate the effect of increased expression of miR-579 on the expression of BAX and CDKN1A 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-579 precursor sequence was cloned into the plasmid. The recombinant structure was transmitted to the cells of HEK293Twith and pMD2 plasmids. Viral particles were concentrated using Ultra Centrifuge. Viral titration was calculated by flow cytometry. The viral particles produced were transferred to the A-172 cell line. Finally, by using Real-Time PCR, changes in expression levels of miR-579 and BAX and CDKN1A genes were investigated.
Findings: The presence of miR-579 gene precursor in the plasmid was confirmed by colony PCR and sequencing methods. The study showed that the level of miR-579 expression in infected cells with the recombinant virus was found to be up-regulated compared to the control group. miR-579 increased the BAX gene expression by three times. But, there was no significant change in the expression of CDKN1A gene expression.
Conclusion: Increased expression of miR-579 in the A-172 cell line could increase the expression of BAX gene. However, the CDKN1A gene expression does not change significantly.
Materials and Methods: In this experimental study, in order to produce recombinant lentiviral vectors, a gene containing the miR-579 precursor sequence was cloned into the plasmid. The recombinant structure was transmitted to the cells of HEK293T
Findings: The presence of miR-579 gene precursor in the plasmid was confirmed by colony PCR and sequencing methods. The study showed that the level of miR-579 expression in infected cells with the recombinant virus was found to be up-regulated compared to the control group. miR-579 increased the BAX gene expression by three times. But, there was no significant change in the expression of CDKN1A gene expression.
Conclusion: Increased expression of miR-579 in the A-172 cell line could increase the expression of BAX gene. However, the CDKN1A gene expression does not change significantly.
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