Apoptosis Induced by Prednisolone Occurs without Altering the Promoter Methylation of BAX and BCL-2 Genes in Acute Lymphoblastic Leukemia Cells CCRF-CEM

Saiedeh Ganbarjeddi,Reza Mousavi Ardehaie,Majid Zamani,Amin Ghasemi,Maryam Hemmatzadeh,Milad Zadi Heydarabad,Sajjad Esmaeili,Shahin Mohammadi,Sina Baharaghdam,Ako Azimi

doi:10.31557/apjcp.2020.21.2.523

Abstract

Objective:one of the main mechanisms in which cancer cells are resistant to chemotherapy drugs and therapeutic strategies is resistance to apoptosis due to these anticancer factors. Regulating the expression of genes through epigenetics, especially regulation through methylation, is one of the key aspects of regulating gene expression and the function of genes, which is also regulated by the pathways regulating the pathway of apoptosis. The epigenetic regulatory phenomenon in cancer cells can undergo a change in regulation and induces resistance to apoptosis against chemotherapy and anticancer factors. The purpose of the present scrutiny was defined to probe the effect of subtoxic prednisolone dose on the level of promoter methylation and gene expression of BAX and BCL2 in the CCRF-CEM cells. Methods:The treated cells by prednisolone, cultured in RPMI 1640 medium in standard condition. Alteration in promoter DNA methylation was analyzed by use of methylation specific-PCR (MSP) technique after the defined intervened time of Prednisolone treatment with a subtoxic dose. Results:Prednisolone can induce apoptosis via alteration in BAX and BCL2 genes, based on our previous scrutiny. This essay shows no varies in the Pattern of DNA methylation of examined genes; however, prednisolone changes the expression of examined genes. Conclusion:Lack of alteration through prednisolone treatment in DNA methylation template of BAX and BCL2 genes make this possible that Prednisolone affects apoptotic gene expression via different pathways, which need more research to be done about it.

Highlights

Acute Lymphoblastic Leukemia (ALL) is the most common type of childhood leukemia, which is generally recognized in marrow and blood through an acute attack and rapid aggregation of immature leukemia cells with a common peak at the age of 2-5 years (Holleman et al, 2004; Azimi et al, 2016)
Regulating the expression of genes through epigenetics, especially regulation through methylation, is one of the key aspects of regulating gene expression and the function of genes, which is regulated by the pathways regulating the pathway of apoptosis
In order to probe the changes on the promoter methylation profile of target genes by prednisolone, the methylation specific-PCR (MSP) technique was applied after 24 and 48 h incubation and the outcomes were evaluated in comparison with untreated cells

Summary

Introduction

Acute Lymphoblastic Leukemia (ALL) is the most common type of childhood leukemia, which is generally recognized in marrow and blood through an acute attack and rapid aggregation of immature leukemia cells (blast) with a common peak at the age of 2-5 years (Holleman et al, 2004; Azimi et al, 2016). One of the principal pathways which make malignant cells resistant to anticancer therapies and therapeutic strategies is resistance to apoptosis. BCL2 family members act as a critical regulator of mitochondrial pathway of apoptosis which potentially consider as target in leukemia therapy. DNA methylation of promotor in GpC island consider as common mechanism for inhibiting the gene expression in cancerous cells (Vahdani et al, 2019; Hervouet et al, 2013). This resistance to cancer cell apoptosis is usually one of the scientists’ major

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