| Objective:Multiple myeloma (MM) is presently an incurable malignant plasma cell tumor. The objective of this study was to investigate expression of interferon regulatory factor (ICSBP/IRF8) and the potential role of DNA methylation in silencing ICSBP/IRF8genes in MM cell line U266and enriched MM cells from10patients.Methods:Using the human MM cell line U266and enriched MM cells from10MM patients. Enriched normal bone marrow mononuclear cells from10healthy persons were collected and used as normal controls. Expression of ICSBP/IRF8genes was investigated by real-time fluorescence quantitative PCR (fetch the CT value, using2-AACT to calculate); Expression of EZH2was fetched by WESTERN-BLOT(using the ratio of purpose protein EZH2and internal reference β-actin expression as the final results).DNA methylation of the interferon consensus sequence-binding protein (ICSBP/IRF8) gene was measured using methylation-sensitive PCR(using the ratio of purpose gene ICSBP/IRF8and internal reference β-actin expression as the final results).Results:Our preliminary in vitro result showed that the MM cell line U266and the MM-BMMNC from10MM patients were found lacking of ICSBP/IRF8gene expression,, but rich in EZH2,associated with hypermethylation of the CpG island in the ICSBP/IRF8promoter, which differences are statistically significant (p<0.05).Conclusion:Expression of the ICSBP/IRF8gene is silenced in the MM-BMMNC and hypermethylation of the CpG island in the ICSBP/IRF8promoter is a frequent event in MM-BMMNC. The ICSBP/IRF8gene silencing caused by DNA methylation may probably take part in the pathogenesis and development of multiple myeloma. |
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