Showing 7 results for Dna Methylation
Ghasemi A, Ghotaslou A, Mohammadi M, Ghaffari K, Abbasian S,
Volume 7, Issue 1 (11-2014)
Abstract
Background: In acute myeloblastic leukemia, a large number of tumor suppressor genes are silenced through DNA
methylation such as CDKN2B & p73. Wnt inhibitory factor 1 (WIF1) and Dickkopf-3 (DKK-1) are negative regulators of
Wnt signaling pathway. In the present study, we evaluated the methylation status of WIF1 and DKK-1 genes in acute
myeloblastic leukemia patients.
Patients and Methods: Blood samples were taken from 120 AML patients and 25 healthy control subjects. DNA was
isolated, treated with sodium bisulphite, and examined using methylation-specific polymerase chain reaction (MSP)
with primers specific for methylated and unmethylated sequences of the WIF1 and DKK-1 genes.
Results: The frequency of aberrant hypermethylation of WIF1 and DKK-1 genes in acute myeloblastic leukemia
patients were determined to be 35% (42/120) and 28.3% (34/120), respectively. In addition, for all subjects in control
group, methylation of WIF1 and DKK-1 genes were negative. Patients with M0 subtype of FAB-AML had the highest
incidence of hypermethylation of WIF1 (P = 0.003) and DKK-1 (P = 0.005) genes.
Conclusion: The present study showed that, like many solid tumors, WIF1 and DKK-1 genes methylation also occurs
in acute myeloblastic leukemia. The study of other antagonists of Wnt signaling pathways are recommended.
Key words: AML, Wnt inhibitory factor 1, dickkopf, DNA methylation.
Ali Ghasemi, Abbas Ghotaslou, Kazem Ghaffari, Mohsen Mohammadi,
Volume 7, Issue 5 (12-2015)
Abstract
Background: Several studies have examined the presence of DNA methylation of CpG islands in leukemia. Methylation of SOX17 and RUNX3 genes may play a role in leukemogenesis through silencing tumor suppressor genes. We investigated the methylation status of SOX17 and RUNX3 genes in patients with acute leukemia.
Methods: In this case-control study, peripheral blood samples from 100 AML and 100 ALL patients and 100 healthy controls were collected. Isolated DNA was treated with sodium bisulfite and methylation status was examined by methylation specific PCR (MS-PCR) with primers specific for methylated and unmethylated sequences of SOX17 and RUNX3 genes.
Results: The frequency of hypermethylation of SOX17 and RUNX3 genes were 36% and 28%I in patients with acute myeloid leukemia (AML), and 21% and 22% in patients with acute lymphoblastic leukemia (ALL), respectively. Aberrant methylation of these genes was found in all FAB classifications of AML and ALL. Hypermethylation of SOX17 (P=0.055) and RUNX3 (P=0.003) genes were associated with FAB-M0 and M1 subtypes of AML, respectively. Also, aberrant methylation of RUNX3 gene was associated with FAB-L1 subtype of ALL (P=0.053). There was not any significant association between hypermethylation of SOX17 and RUNX3 genes and clinical parameters of patients with leukemia including sex, age, WBC, and platelet counts.
Conclusion: Hypermethylation of SOX17 and RUNX3 genes was seen in patients with acute leukemia. Moreover, no significant association was observed between hypermethylation of SOX17 and RUNX3 and induction of remission.
Hamid Gholipour, Saeid Abroun, Mehrdad Noruzinia, Sasan Ghaffari, Amirhosein Maali, Mehdi Azad,
Volume 10, Issue 4 (12-2018)
Abstract
Background: Epigenetic modifications, such as methylation can occur in multiple myeloma. SMG1is an important gene involved in cell growth which defect in methylation of its promoter leads to reduction of cell apoptosis and uncontrolled proliferation. In this study, we identified the methylation status of the SMG1 gene promoter in patients with multiple myeloma.
Methods: Methylation status of SMG1 promoter in 9 patients with multiple myeloma and 4 healthy subjects as control was determined by Methylation-specific PCR (MSP) method.
Results: SMG1 promoter in all myeloma patients was hemi-methylated. Meanwhile, in healthy subjects, two cases were hemi-methylated and the other two were normal.
Conclusion: The results of this study indicated that the prevalence of SMG1 promoter methylation in patients with multiple myeloma was higher than general population which could be important in understanding the pathogenesis of the disease.
Kazem Ghaffari, Ali Ghasemi, Mohsen Mohammadi, Sadegh Abbasian,
Volume 13, Issue 1 (3-2021)
Abstract
Background: DNA methylation patterns are often changed in cancer cells. Many of the tumor inhibitor genes are silenced by methylation, such as CDKN2B, p73, and the suppressor of cytokine signaling in patients with acute myeloblastic leukemia (AML). Secreted frizzled-related protein -4 and -5 (SFRP4, 5) are negative regulators of the Wnt signaling pathway. We aimed to evaluate the methylation status of SFRP4 and SFRP5 genes in patients with AML.
Methods: Blood samples were isolated from 60 patients with AML and 30 healthy controls. DNA was exploited, treated with sodium bisulfite, and tested utilizing methylation-specific polymerase chain reaction with specific primers for methylated and unmethylated sequences of the SFRP4 and SFRP5 genes.
Results: The frequency of unfit hypermethylation of SFRP4 and SFRP5 genes in patients with AML was characterized to be 50% (30/60) and 40% (24/60), respectively. Moreover, for all the subjects in the control group, methylation of SFRP4 and SFRP5 genes was negative. The spread of SFRP4 and SFRP5 promoter methylation in patients with AML was higher than the control population.
Conclusion: Hypermethylation was seen in SFRP4 and SFRP5 genes in patients with AML.
Saeid Kaviani, Aryan Salahi-Niri, Mohammad Hossein Mohammadi, Mohsen Hamidpour, Shadi Esmaeili,
Volume 16, Issue 1 (3-2024)
Abstract
Background: Epigenetics is crucial in differentiating mesenchymal stem cells (MSCs) into adipocytes. Specifically, DNA methylation, an epigenetic modification, regulates the expression of genes involved in this process. The peroxisome proliferator-activated receptor gamma (PPARγ) gene is a critical player in adipocyte differentiation, with epigenetic changes affecting its expression.
Methods: We isolated mesenchymal stem cells (MSCs) from the human bone marrow. The isolated MSCs were expanded and cultured in a differentiation medium for two weeks. DNA extraction was performed on undifferentiated and differentiated adipocytes after the culturing process. The methylation status of the promoter region of the PPARγ gene was assessed using methylation-specific primers (M for methylated and U for unmethylated) in a methylation-specific PCR (MSP) assay. This analysis involved the treatment of DNA samples with sodium bisulfite to convert unmethylated cytosine to uracil, thereby enabling the differentiation between methylated and unmethylated regions of the gene.
Results: The successful differentiation of MSCs into adipocytes was confirmed by the accumulation of lipid droplets within the differentiated cells, as visualized by the oil Red O dye staining. This observation provides strong evidence of the commitment of MSCs towards the adipogenic lineage and their ability to undergo adipocyte differentiation. Surprisingly, the MSP analysis revealed no significant changes in the methylation pattern of this gene following differentiation. The PPARγ gene promoter region exhibited an unmethylated status in both undifferentiated and differentiated states.
Conclusion: Our study revealed that additional genetic or epigenetic mechanisms control the expression of PPARγ during the adipogenic differentiation of mesenchymal stem cells. These findings highlight the regulatory role of PPARγ in the differentiation pathway from mesenchymal stem cells to adipocytes.
Saeed Turkmen, Neda Karami Chermahini, Amirhosein Maali, Mohammad Reza Keramati, Mohammad Hossein Ahmadi, Mehdi Azad, Samaneh Borouman-Noughabi,
Volume 16, Issue 3 (9-2024)
Abstract
Background: Aberrant DNA methylation is a key epigenetic alteration observed in multiple cancers. Acute myeloid leukemia (AML), a prominent form of hematopoietic cancer, is characterized by abnormal proliferation and differentiation of myeloid progenitor cells. This study focuses on examining the methylation status of the CpG islands in the DNMT1 and CDX2 promoter regions and exploring their correlation with prognostic hematological laboratory parameters across three phases of AML: newly diagnosed, undergoing treatment, and in remission.
Material and methods: This follow-up case-control study recruited 11 new cases of confirmed AML admitted to Shariati Hospital in Tehran. All patients received AML treatment according to FDA protocol. The samples (peripheral blood) were collected before medication (new case phase), during medication (under treatment phase), and in the remission phase. Then, genomic DNA was extracted and treated with the bisulfite treatment method. Then, methylation-specific PCR (MSP) was conducted to amplify treated DNAs using two methylated and unmethylated primers related to their promoters' DNMT1 and CDX2 CpG- islands. All statistical analysis was performed using SPSS v.25.
Results: The results of the methylation pattern of DNMT1 gene promoter CpG islands in the present study show that the hemimethylated pattern of the DNMT1 gene promoter is predominant in control (100%), new case phase (90.9%), under treatment phase (72.7%), and remission phase (100%). In the case of the CDX2 gene, the unmethylated pattern is predominant in control (57.14%), new case phase (72.7%), under-treatment phase (90.9%), and remission phase (81.8%). These differences were not statistically significant. No methylated pattern was observed in the control group, and different phases of AML were used for DNMT1 and CDX2. Also, the methylation status of DNMT1 and CDX2 were not correlated with prognostic hematological laboratory parameters.
Conclusion: The methylation patterns of CDX2 and DNMT1 are not different in healthy individuals and AML patients, as well as in different phases of AML. Also, the methylation patterns of CDX2 and DNMT1 cannot help determine the prognosis of AML patients through changes in hematological laboratory parameters.
Seied Rasoul Razavi Babaheidari, Aryan Salahi-Niri, Mohammad Hossein Mohammadi, Mohsen Hamidpour, Shadi Esmaeili,
Volume 17, Issue 1 (3-2025)
Abstract
Background: Lipoprotein lipase (LPL) is a critical enzyme in lipid metabolism that hydrolyzes triglyceride-rich lipoproteins. While its role in mature adipose tissue is well understood, the epigenetic regulation of LPL during mesenchymal stem cell (MSC) differentiation into adipocytes remains poorly characterized. This study aimed to investigate the temporal expression pattern of LPL and its relationship with DNA methylation during adipogenic differentiation of human bone marrow-derived MSCs.
Methods: Human bone marrow MSCs were isolated and characterized using flow cytometry for specific surface markers (CD34, CD31, CD90, and CD105). Cells were differentiated into adipocytes over 14 days and osteoblasts over 21 days using specific differentiation media. Differentiation was confirmed through Oil Red O and Alizarin Red staining respectively. LPL gene expression was analyzed using both qualitative RT-PCR and quantitative real-time PCR at day 0 (undifferentiated) and day 14 (differentiated) timepoints. DNA methylation patterns were assessed using methylation-specific PCR (MSP) following bisulfite conversion, with collagen gene serving as an internal control.
Results: Flow cytometry confirmed MSC identity through positive expression of CD166, CD13, CD105, and CD44. Successful adipogenic differentiation was demonstrated by Oil Red O-positive lipid droplet accumulation, while osteogenic differentiation was confirmed by Alizarin Red S staining of calcium deposits. LPL gene expression was absent in undifferentiated MSCs but showed significant expression in differentiated adipocytes at day 14, coinciding with morphological changes and lipid accumulation.
Conclusion: This study demonstrates that LPL expression is epigenetically regulated during MSC differentiation into adipocytes, with significant changes in both gene expression and DNA methylation patterns. The temporal correlation between LPL expression, methylation status, and adipogenic differentiation suggests that LPL serves as a key molecular switch in this process.
