甲基化修饰致PCDH10基因在多发性骨髓瘤中沉默  

Methylation induces protocadherin-10 silence in multiple myeloma

作　　者：李颖[1] 陈建斌[1] 宋君君[1] 杨泽松[2] 向廷秀[3] 黄宗干[1] 

机构地区：[1]重庆医科大学附属第一医院血液科,重庆400016 [2]重庆医科大学附属第一医院血液科实验室,重庆400016 [3]重庆医科大学附属第一医院分子肿瘤及表观遗传学实验室,重庆400016

出　　处：《第三军医大学学报》2012年第9期817-820,共4页Journal of Third Military Medical University

基　　金：重庆市卫生局2011年医学科研计划重点项目(2011-1-032);重庆市医学科研计划(2010-2-108)~~

摘　　要：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。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,and may be involved in the incidence of myeloma.

关 键 词：多发性骨髓瘤 PCDH10 DNA甲基化 

分 类 号：R394-33[医药卫生—医学遗传学]