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[Silencing of ICSBP/IRF8 expression in U226 cells and bone marrow mononuclear cells from patients with multiple
Gu Jia1, Yan-Ping Ma, Ling Zhang
1Department of Hematology, Shanxi Medical University Second Hospital, Taiyuan, Shanxi Province, China.
Abstract:
The objective of this study was to investigate expression of interferon regulatory factors (ICSBP/IRF8) and the potential role of DNA methylation in silencing ICSBP/IRF8 gene in multiple myeloma (MM) cell line U266 and bone marrow mononuclear cells from 10 MM patients (MM-BMMNC). The bone marrow mononuclear cells from 10 healthy persons (N-BMMNC) were collected and used as normal controls. Expression of ICSBP/IRF8 gene was detected by real-time fluorescence quantitative PCR (using 2(-ΔΔCT) to calculate); DNA methylation level of the ICSBP/IRF8 gene was measured using methylation-specific PCR (using the ratio of interest gene ICSBP/IRF8 and internal reference β-actin expression as results). The results showed that as compared with N-BMMNC the lower expression of ICSBP/IRF8 gene was found in U266 cells and MM-BMMNC, the hypermethylation of the CpG island in the ICSBP/IRF8 promoter was observed, there were significant differences between N-BMMNC and MM-BMMNC or U266 cells (P < 0.05). It is concluded that the expression of ICSBP/IRF8 gene can be silenced in the MM-BMMNC and U226 cells. As the hypermethylation of CpG island in ICSBP/IRF8 promoter is a frequent event in MM cells, the ICSBP/IRF8 gene silencing caused by DNA methylation may take part in the pathogenesis and development of MM.
Insights
Multiple myeloma cells show reduced interferon regulatory factor 8 (IRF8) gene expression. DNA hypermethylation in the IRF8 promoter likely silences this gene, contributing to multiple myeloma development.
Area of Science:
- Molecular Biology
- Cancer Genetics
- Epigenetics
Context:
- Multiple myeloma (MM) is a hematological malignancy characterized by uncontrolled plasma cell proliferation.
- Interferon regulatory factors (IRFs) play crucial roles in immune regulation and cellular differentiation.
- Aberrant gene expression and epigenetic modifications are implicated in MM pathogenesis.
Purpose:
- To investigate the expression levels of interferon regulatory factor 8 (IRF8) in multiple myeloma.
- To explore the role of DNA methylation in the silencing of the IRF8 gene in MM.
- To compare IRF8 expression and promoter methylation between MM cells and normal controls.
Summary:
- Lower expression of the IRF8 gene was observed in the U266 MM cell line and bone marrow mononuclear cells (MM-BMMNC) from MM patients compared to normal controls (N-BMMNC).
- Hypermethylation of the CpG island within the IRF8 promoter region was detected in U266 cells and MM-BMMNC.
- Significant differences in IRF8 expression and promoter methylation were found between MM cells and normal controls (P < 0.05).
Impact:
- The findings suggest that IRF8 gene silencing, mediated by DNA methylation, is a frequent event in MM.
- This epigenetic silencing of IRF8 may contribute to the pathogenesis and progression of multiple myeloma.
- Understanding IRF8 regulation in MM could reveal novel therapeutic targets for epigenetic-based treatments.
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