儿童非霍奇金淋巴瘤ZO-1基因甲基化状态分析及临床意义  

ZO-1 gene methylation status and its clinical significance in children with nonHodgkin lymphoma

作　　者：刁玉巧[1] 曲凡[1] 杨明娟[1] 孟建辉[1] 朱秀丽[1] 陈健[1] 

机构地区：[1]河北医科大学第四医院儿科,河北石家庄050011

出　　处：《中国当代儿科杂志》2014年第6期619-623,共5页Chinese Journal of Contemporary Pediatrics

基　　金：河北省科技支撑计划项目(12277753)

摘　　要：目的探讨紧密连接蛋白(ZO-1)基因启动子区甲基化状态在儿童非霍奇金淋巴瘤(NHL)Ⅳ期检测中的临床意义,以进一步寻找病因及早期诊断方法。方法将55例确诊为NHLⅣ期(T细胞型40例,B细胞型15例)患儿的骨髓标本作为病例组,20例非血液肿瘤患儿骨髓标本作为对照组。采用甲基化特异性PCR法(MS-PCR)检测病例组与对照组骨髓中ZO-1基因启动子区甲基化状态,并检测光密度积分值(IOD)。逆转录PCR法(RT-PCR)检测病例组与对照组ZO-1基因mRNA的表达情况。结果 MS-PCR检测结果显示病例组治疗前39例出现ZO-1基因甲基化条带,阳性率为70.9%(39/55),对照组20例均呈非甲基化状态;治疗过程中,行动态观察NHL患儿47例,其中T细胞型32例,B细胞型15例,治疗前ZO-1基因甲基化阳性率分别为72%和67%,两者差异无统计学意义(P>0.05)。T细胞型与B细胞型NHL患儿治疗前与化疗早期、中期比较差异均无统计学意义(P>0.05),而与化疗后期比较差异均有统计学意义(P<0.05)。RT-PCR检测结果显示病例组中ZO-1甲基化阳性病例均无mRNA表达,对照组患儿均表达ZO-1 mRNA。外周血白细胞总数与ZO-1基因IOD值无直线相关关系(r=0.093,P=0.575);骨髓中恶性肿瘤细胞数与ZO-1基因IOD值呈正相关关系(r=0.669,P<0.001)。结论 ZO-1基因在儿童NHL中呈特异性高甲基化状态,其程度与骨髓中恶性肿瘤细胞数呈正相关。ZO-1基因可能成为新的分子标志,为临床早期诊断、疗效判断、预后评价、微小残留检测提供新的指标。Objective To investigate the methylation status of zonula occludens-1（ZO-1） gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma（NHL） and to provide a basis for further etiological study and early diagnosis of this disease.Methods Fifty-five children with a confirmed diagnosis of stage IV NHL（40 cases of T-NHL and 15 cases of B-NHL） were selected as the case group,and 20 children with diseases other than hematologic malignancies were selected as the control group.Bone marrow samples were collected from these subjects.Methylation-specific PCR（MS-PCR） was applied to evaluate the methylation status of ZO-1 gene promoter,and the integrated optical density（IOD） was determined.RT-PCR was used to measure the mRNA expression of ZO-1.Results MS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39（70.9%） of 55 patients in the case group before treatment,while no ZO-1 gene promoter methylation was detected in the control group.With close tracking of 47 cases in the study group,consisting of 32 cases of T-NHL and 15 cases of B-NHL,the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%,respectively,（P＆gt;0.572）.The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy（P＆gt;0.05）,but they showed significant changes in methylation rate in the late phase of chemotherapy（P＆lt;0.05）.RT-PCR showed that the

关 键 词：ZO-1基因 甲基化 非霍奇金淋巴瘤 儿童 

分 类 号：R733.1[医药卫生—肿瘤]