Analysis of p53 tumour suppressor gene somatic mutations in rheumatoid arthritis synovium
Analysis of p53 tumour suppressor gene somatic mutations in rheumatoid arthritis synovium
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DOI：
10.1093/rheumatology/39.3.262
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发表时间：
2000-03-01
期刊：
影响因子：
5.5
通讯作者：
Hayashi, K
中科院分区：
文献类型：
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作者：
Inazuka, M;Tahira, T;Hayashi, K
Objective.为探讨抑癌基因p53在类风湿关节炎（RA）滑膜增殖中的作用，我们检测了RA患者滑膜成纤维样B型滑膜细胞中p53基因的突变。滑膜成纤维细胞样B型滑膜细胞从9名日本RA患者的滑膜组织中制备。采用聚合酶链反应（PCR）产物后标记的突变检测方法对p53基因4-11外显子的cDNA区域进行突变筛查，并采用单链构象多态性条件进行自动毛细管电泳分析，然后对PCR产物的亚克隆进行直接测序。9例RA患者中4例（44.4%）检测到p53基因突变并可能伴有功能改变。在总共262个p53 cDNA亚克隆中，10个亚克隆携带18个p53突变，其中8个与氨基酸改变或蛋白质截短相关。在p53功能突变中，在3个亚克隆中的2名患者中鉴定出氨基酸残基245处Gly替换为Asp（G245 D）。G245 D是在不同RA个体中反复鉴定的第一个突变。G245 D也是人类肿瘤中相对常见的突变之一。在某些RA患者中，p53功能障碍可能在滑膜组织增殖中起作用。G245 D突变可能特别需要进一步研究，因为它是RA中第一个反复鉴定的p53突变，也是人类癌症中经常鉴定的突变之一。
Objective. In order to study the role of the p53 tumour suppressor gene in the proliferation of rheumatoid arthritis (RA) synovium, we analysed the mutation of p53 in the synovial fibroblast-like type B synoviocyte from RA patients.Methods. Synovial fibroblast-like type B synoviocytes were prepared from the synovial tissues from nine Japanese patients with RA. The p53 cDNA region from exons 4-11 was screened for mutations by the streamlined mutation detection method in which polymerase chain reaction (PCR) products are post-labelled and are analysed by automated capillary electrophoresis using single-strand conformation polymorphism conditions, followed by direct sequencing of the subclones of the PCR products.Results. p53 mutation with possible functional alteration was detected in four of the nine RA patients (44.4%). Of a total of 262 p53 cDNA subclones, 10 subclones were carrying 18 p53 mutations, eight of which were associated with amino acid alterations or protein truncation. Of the p53 functional mutations, a substitution of Gly at amino acid residue 245 to Asp (G245D) was identified in two patients in three subclones. G245D was the first mutation that was recurrently identified in different RA individuals. G245D is also one of the relatively common mutations in human cancers.Conclusions. In some patients with RA, dysfunction of p53 might play a role in the proliferation of the synovial tissue. G245D mutation might especially need further study as it is the first recurrently identified p53 mutation in RA and is also one of the frequently identified mutations in human cancers.