Diao Yu-Qiao, Qu Fan, Yang Ming-Juan, Meng Jian-Hui, Zhu Xiu-Li, Chen Jian
Zhongguo Dang Dai Er Ke Za Zhi. 2014 Jun;16(6):619-23.
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.
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.
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>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>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).
ZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.
探讨闭合蛋白-1（ZO-1）基因启动子甲基化状态及其在IV期非霍奇金淋巴瘤（NHL）患儿中的临床意义，为该疾病的进一步病因学研究及早期诊断提供依据。
选取确诊为IV期NHL的55例患儿（T-NHL 40例，B-NHL 15例）作为病例组，选取20例非血液系统恶性肿瘤患儿作为对照组。采集这些受试者的骨髓样本。采用甲基化特异性PCR（MS-PCR）评估ZO-1基因启动子的甲基化状态，并测定积分光密度（IOD）。采用逆转录聚合酶链反应（RT-PCR）检测ZO-1的mRNA表达。
MS-PCR显示，病例组55例患者中39例（70.9%）治疗前ZO-1基因启动子出现甲基化条带，而对照组未检测到ZO-1基因启动子甲基化。对研究组47例患者（T-NHL 32例，B-NHL 15例）进行密切随访，治疗前ZO-1基因启动子甲基化率分别为72%和67%，（P>0.572）。T-NHL和B-NHL病例在化疗早、中期甲基化率无明显变化（P>0.05），但在化疗后期甲基化率有明显变化（P<0.05）。RT-PCR显示，携带甲基化ZO-1基因的NHL病例无ZO-1的mRNA表达，而对照组所有患儿均有ZO-1的mRNA表达。外周血白细胞总数与ZO-1基因IOD之间无线性关系（r=0.093，P=0.575）；骨髓中恶性细胞数量与ZO-1基因IOD呈正相关（r=0.669，P<0.001）。
NHL患儿中ZO-1基因呈高甲基化状态，甲基化水平与骨髓中恶性细胞数量呈正相关。ZO-1可作为早期诊断、疗效评估、预后评价及微小残留病检测的新型分子标志物。
