Li Ying,Chen Jianbin,Song Junjun,Yang Zesong,Xiang Tingxiu,Huang Zonggan.Methylation induces protocadherin-10 silence in multiple myeloma[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(09):817-820.
Li Ying,Chen Jianbin,Song Junjun,Yang Zesong,Xiang Tingxiu,Huang Zonggan.Methylation induces protocadherin-10 silence in multiple myeloma[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(09):817-820.DOI:
Methylation induces protocadherin-10 silence in multiple myeloma
目的 研究PCDH10基因在多发性骨髓瘤(multiple myeloma,MM)患者及细胞系KM3、RPMI-8226中的甲基化状态,分析甲基化对其沉默的影响。 方法 常规骨髓穿刺术获取MM患者骨髓样本,培养骨髓瘤细胞系KM3、RPMI-8226,以增生性骨髓象骨髓样本作为对照。RT-PCR检测其中PCDH10基因表达水平,甲基化特异性PCR(methylation-sensitive PCR,MSP)法检测PCDH10基因甲基化状态,通过5-氮杂-2′-脱氧胞苷和组蛋白去乙酰化酶抑制剂[5-aza-2′-deoxycytidine (Aza) and trichostatin A (TSA),A+T]处理MM细胞系,检测药物处理前后PCDH10甲基化状态及表达水平。 结果 PCDH10基因在正常人群中表达,但在MM患者骨髓及细胞系中广泛低表达,且77.27%(34/44)的MM患者PCDH10呈甲基化状态。PCDH10基因可经A+T试验逆转甲基化状态并恢复表达。 结论 DNA甲基化可能在多发性骨髓瘤的PCDH10基因沉默中发挥作用,从而参与骨髓瘤的发生。
Abstract
Objective To determine the methylation status of protocadherin-10 (PCDH10) in multiple myeloma (MM) patients and MM cell lines KM3 and RPMI-8226
and investigate the effect of its methylation on PCDH10 silence. Methods A total of 44 MM tissue samples from January to December 2010 hospitalized in our department were collected. Bone marrow tissues of 9 age- and sex-matched patients with mild anemia obtained by conventional bone marrow biopsy served as control. The expression of PCDH10 gene was detected by RT-PCR in above tissue samples
and multiple myeloma cell lines KM3 and RPMI-8226. Methylation-sensitive PCR (MSP) was used to test the methylation of PCDH10 CpG islands. Then
after the KM3 and RPMI-8226 cells were treated with 5-aza-2′-deoxycytidine and trichostatin A (Aza and TSA
A+T)
the changes of PCDH10 were studied by RT-PCR and MSP. Results PCDH10 was expressed well in normal population while extensive mildly expressed in MM patients and cell lines. Among the 44 patients (23 females
21 males)
77.27% (34/44) whom presented methylation of PCDH10. Furthermore
A+T restored PCDH10 expression in myeloma cell lines. Conclusion DNA methylation may play a key role in the silence of PCDH10 gene in multiple myeloma