CAJ | 학술논문

目的　探讨SHP-1基因异常甲基化在骨髓增生异常综合征(MDS)发生、发展中的作用和相关机制.方法　将63例MDS患者根据IPSS积分系统分为较低危和较高危组.采用甲基化特异性PCR(MS-PCR)对MDS细胞系SKK-1细胞和MDS患者骨髓细胞进行SHP-1基因启动子甲基化状态的检测;用Western blot方法检测正常对照者和患者骨髓单个核细胞和SKK-1细胞磷酸化信号传导和转录激活因子3( p-STAT3)蛋白的表达.结果　SKK-1细胞存在SHP-1基因启动子部分甲基化,正常对照者骨髓细胞均为非甲基化,较高危组MDS患者SHP-1启动子甲基化率显著高于较低危组(63.3％对24.2％)(P＜0.05);按WHO分型中染色体核型和骨髓原始细胞比例分组,SHP-1启动子甲基化率在RAEB-Ⅱ、核型差组和骨髓原始细胞占0.11 ～0.19组显著增高(P＜ 0.05);SKK-1细胞中存在p-STAT3蛋白的表达,较高危组MDS患者p-STAT3蛋白的表达显著高于较低危组(66.7％对18.2％);相关分析显示SHP-1启动子甲基化的出现与p-STAT3蛋白的表达有相关性.结论　SHP-1基因启动子的异常甲基化在MDS的发病和疾病进展中发挥作用,其机制可能涉及STAT3信号通路的激活,检测MDS患者SHP-1启动子甲基化状态有助于对疾病预后的判断.
목적　탐토SHP-1기인이상갑기화재골수증생이상종합정(MDS)발생、발전중적작용화상관궤제.방법　장63례MDS환자근거IPSS적분계통분위교저위화교고위조.채용갑기화특이성PCR(MS-PCR)대MDS세포계SKK-1세포화MDS환자골수세포진행SHP-1기인계동자갑기화상태적검측;용Western blot방법검측정상대조자화환자골수단개핵세포화SKK-1세포린산화신호전도화전록격활인자3( p-STAT3)단백적표체.결과　SKK-1세포존재SHP-1기인계동자부분갑기화,정상대조자골수세포균위비갑기화,교고위조MDS환자SHP-1계동자갑기화솔현저고우교저위조(63.3％대24.2％)(P＜0.05);안WHO분형중염색체핵형화골수원시세포비례분조,SHP-1계동자갑기화솔재RAEB-Ⅱ、핵형차조화골수원시세포점0.11 ～0.19조현저증고(P＜ 0.05);SKK-1세포중존재p-STAT3단백적표체,교고위조MDS환자p-STAT3단백적표체현저고우교저위조(66.7％대18.2％);상관분석현시SHP-1계동자갑기화적출현여p-STAT3단백적표체유상관성.결론　SHP-1기인계동자적이상갑기화재MDS적발병화질병진전중발휘작용,기궤제가능섭급STAT3신호통로적격활,검측MDS환자SHP-1계동자갑기화상태유조우대질병예후적판단.
Objective To explore the role of SHP-1 promoter methylation on the pathogenesis and　progression in myelodysplastic syndromes (MDS) and its related mechanism.Methods 63 MDS patients　were divided into low-grade(LG) group and high-grade(HG) group according to IPSS score system.Bone　marrow samples were collected. Methylation specific-PCR (MSP) were used to detect the status of SHP-1　promoter methylation in bone marrow (BM) samples from different risk MDS patients and MDS cell line,SKK-1.Western blot was used to detect signal transduction and activator of transcription(STAT3) activation in SKK-1 cell line and MDS patients.Results No SHP-1 promoter methylation could be detected in healthy controls BM.Partially methylation was found in SKK-1 cell line.Methylation rate of SHP-1 gene promoter was found in BM of 24.2％ of low-grade MDS patients and 63.3％ of high-grade MDS patients,the difference between these two groups was statistically significant (P ＜ 0.05) ; Patients were divided into different groups according to WHO subtype,chromosomal karyotype and blast cells in bone marrow,methylation rates of SHP-1 were significantly higher in RAEB- Ⅱ,poor karotype group and samples with 0.11 - 0.19 blast cells ( P ＜0.05 ) ;The phosphorylation protein of STAT3 was detected in SKK-1 cell line.The expression of phosphorylation STAT3 was significantly higher in HG group than in LG group (66.7％ vs 18.2％ ) (P ＜0.05).There was a significant correlation between SHP-1 promoter methylation and STAT3 phosphorylation.Conclusion　Abnormal methylation of SHP-1 gene promoter might have tentative role in the pathogenesis and progression of　MDS,which may be involved in STAT3 activation.Detection of SHP-1 promoter methylation may be helpful　to evaluate the prognosis of MDS.