摘要
目的探讨紧密连接蛋白(zonula occluden protein-1,ZO-1)基因启动子区甲基化状态在儿童急性白血病(AL)骨髓检测中的临床意义。方法将48例确诊为AL(35例急性淋巴细胞白血病,13例急性髓细胞白血病)患儿的骨髓标本作为研究组,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(MS-PCR)检测研究组与对照组ZO-1基因启动子区甲基化状态。逆转录PCR(RT-PCR)检测研究组与对照组ZO-1基因mRNA的表达情况。结果 MS-PCR检测结果显示,治疗前研究组ZO-1基因甲基化阳性率为68.8%(33/48),对照组均呈非甲基化状态(P<0.05);动态观察31例急性淋巴细胞白血病ZO-1基因甲基化阳性率:治疗前64.5%(20/31),治疗第33天48.4%(15/31),治疗第12周29.0%(9/31)。治疗过程中复发2例,甲基化状态由部分甲基化条带转变为完全甲基化条带。5例疗程结束,停药前仍有1例可检出。治疗前与治疗第33天、治疗第12周比较差异均有统计学意义(P<0.05),治疗第33天与治疗第12周比较差异有统计学意义(P<0.05);急性淋巴细胞白血病68.6%(24/35),与急性髓细胞白血病69.2%(9/13)比较差异无统计学意义(P>0.05)。RT-PCR检测结果显示,研究组治疗前及动态观察中,ZO-1基因甲基化阳性病例均无mRNA表达,对照组全部表达。结论 ZO-1基因启动子区高甲基化状态在儿童急性白血病中有较高的特异性,与疾病发生、发展及转归密切相关。ZO-1基因有可能作为新的白血病分子标志,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 ZO-1 gene promoter in children acute leukemia（AL） and its significance. 【Methods】 48 cases that had diagnosed AL（35 cases of ALL, 13 cases of AML） were selected as study groups, 20 non-blood tumor cases were selected as control groups. Methylation specific PCR（MS- PCR） method was used to detect the bone marrow specimen in study group and control group. Transcription-Polymerase Chain Reaction（RT-PCR） method was used to detect the expression of methylated ZO-1 gene mRNA in study group and control group. 【Results】 MS-PCR showed that 68.8% AL were found ZO-1 gene promoter in bone marrow harboring aberrant methylation, no ZO-1 gene promoter methylation was detected in control group（P〈0.05）. 31 cases of ALL were dynamically observed： the methylation-positive rate before treatment was 64.5%, than the rate became 48.4% in 33 days and 29.0% in 12weeks respectively. 2 patients recurrence, the ZO-1 gene become complete methylation status from partial methylation.1 case was still could be checked out in 5 patients after treatment. There was significant difference between the Pre-treatment and in day 33 or in week 12（P〈0.05）. It was also significant difference between Day 33 and week 12（P〈0.05）; There was no significant difference between ALL（68.6%） and AML（69.2%）. RT-PCR showed that before chemotherapy treatment and dynamic observation, all of those that carried methylated ZO-1 gene had no expression of mRNA, by comparison to the controlled group which all had expression of mRNA. 【Conclusion】 ZO-1 gene shows specific hyper-methylationstatus in children AL, and it is closely related to the occurrence, development and outcome of the disease. It may be a new molecular marker, which may provide an index for early clinical diagnosis, determining efficacy, prognosis analysis and detection of minimal residual disease.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2014年第17期39-43,共5页
China Journal of Modern Medicine
基金
河北省科技支撑计划项目(No:12277753)
