摘要
【目的】探讨JAK/STAT信号转导途径负调控子SHP-1基因启动子区域CpG岛异常甲基化在B细胞淋巴瘤中的意义。【方法】收集存档石蜡包埋组织标本61例(52例B细胞非霍奇金淋巴瘤标本,9例良性增生淋巴结标本),健康人外周血单个核细胞DNA标本15例,用甲基化特异性PCR(methylation-specific PCR,MSP)和非甲基化特异性PCR(unmethylation-specific PCR,un-MSP)检测SHP-1启动子区域CpG岛甲基化状态,MSP、un-MSP和RT-PCR方法分别检测接受或未接受去甲基化处理的Burkitt淋巴瘤细胞系Raji的甲基化状态及mRNA的表达,MTT法检测接受去甲基化干预后细胞生长受抑情况。【结果】SHP-1基因启动子区域在弥漫性大B细胞淋巴瘤和滤泡性淋巴瘤甲基化频率分别为94%及97%,对照组9例淋巴结良性增生标本和15例正常人外周血单个核细胞标本中SHP-1基因启动子区域甲基化频率为0。经去甲基化干预后,Raji细胞SHP-1基因启动子区域呈去甲基化状态,基因恢复表达,细胞生长受到抑制。【结论】SHP-1基因启动子区域启动子区域CpG岛在B细胞淋巴瘤中存在高度甲基化,由其所致的SHP-1基因沉默可能是B细胞淋巴瘤发生的一个重要因素,SHP-1基因的甲基化可作为一个良好的分子诊断标记及可能的治疗靶点。
[Objective] To investigate the methylation state of CpG island in the SHP-1 gene promoter which is one of the negative regulators of JAK/STAT signaling pathway and to evaluate its significance in B-cell lymphoma. [Methods] Sixty-one paraffin specimens including 52 specimens of B-cell non-Hodgkin lymphoma and 9 specimens of benign lymphnodes proliferation and 15 specimens of blood mononuclear cells from health individuals were studied, methylation state of CpG island in SHP-1 gene in the specimens and Raji cell line were detected by methylation specific PCR（MSP） and unmethylation-specific PCR（un-MSP）. The expression level of SHP-1 in Raji cells with or without 5-aza-2＇ -deoxyoytidine treatment were detected by reverse transcriptase PCR （RT-PCR）.The inhibitory ratio of Raji cells were measured by MTT assay. [Results] The frequency of methylation in SHP-1 gene promoter in large B-cell lymphoma（DLBCL） and follicular lymphoma（FL） were 94% and 97%,respectively.In control group, however,9 specimens of benign lymphnodes proliferation and 15 specimens of blood mononuclear cells from health individuals showed no methylation in CpG island of SHP-1 gene promotor （P〈 0.0001 ）. The expression of SHP-1 was recovered and the cell growth was inhibited when the cell exposed to the demethylating reagent. [Conclusion] SHP-1 gene silencling with aberrant CpG methylation is probably one of the critical events to the onset of B-cell lymphoma as well as important implications for the diagnostic markers and the target of gene therapy.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2007年第1期92-96,共5页
Journal of Sun Yat-Sen University:Medical Sciences
