恶性血液病EB病毒RPMS1基因多态性分析及意义

SIGNIFICANCE OF EPSTEIN-BARR VIRUS RPMS1 GENE POLYMORPHISM IN PATIENTS WITH HEMATOLOGICAL MALIGNANCY

目的了解青岛地区EB病毒(EBV)阳性恶性血液病病人RPMS1基因变异特征,并探讨RPMS1变异与恶性血液病发生发展的关系。方法选取青岛地区恶性血液病病人345例和健康成人93例为研究对象,采用实时荧光定量PCR测定EBV DNA拷贝数;采用巢式PCR和DNA测序技术,检测EBV阳性恶性血液病病人和健康成人外周血RPMS1序列,根据其特征性突变和系统进化树对基因变异进行分类,并与同一地区健康人群RPMS1变异类型的分布进行比较。结果有218例EBV阳性标本完成测序分析,共发现3种RPMS1亚型,即RPMS1-A、RPMS1-B和RPMS1-C。RPMS1-A型为主要亚型,在172例标本中检测到,其基因序列高度保守,除个别散在突变外基本与标准株B95-8一致;RPMS1-C相对较少,以D51N突变为特征;RPMS1-B变异型最少,分别以S65N突变和S98G突变为特征。结论青岛地区EBV阳性恶性血液病和健康人群RPMS1基因型均以RPMS1-A型为主,而RPMS1-B和RPMS1-C变异型的地域性分布及其与恶性血液病的确切关系尚不清楚。

Objective To investigate the characteristics of RPMS1gene variation in patients with Epstein-Barr virus(EBV)-positive hematological malignancy in Qingdao,China and the association of RPMS1variation with the development and progression of hematological malignancy.Methods A total of 345patients with hematological malignancy and 93healthy adults in Qingdao were selected as subjects.Quantitative real-time PCR was used to measure EBV DNA copy number,and then nested PCR and DNA sequencing were used to determine the sequence of RPMS1in peripheral blood of the patients with EBV-positive hematological malignancy and healthy adults.The gene variations were classified according to their characteristic mutations and phylogenetic trees,and the distribution of RPMS1variants was compared between the patients with EBV-positive hematological malignancy and healthy adults.Results Sequencing was performed for 218EBV-positive specimens,and three subtypes of the RPMS1gene were found,i.e.,RPMS1-A,RPMS1-B,and RPMS1-C.RPMS1-A was a major subtype of RPMS1and was detected in 172specimens;its gene sequence was highly conserved and was basically consistent with the standard strain B95-8except for a few sporadic mutations.RPMS1-C was less common than RPMS1-A and was characterized by D51N mutation.RPMS1-B was the least common subtype and was characterized by S65N mutation and S98G mutation.Conclusion RPMS1-A is the major subtype of RPMS1in patients with EBV-positive hematological malignancy and health adults in Qingdao,and further studies are needed to clarify the regional distribution of RPMS1-B and RPMS蜏S1-C variants and their association with hematological malignancy.