H-ras oncogene point mutations in arthritic synovium.
H-ras oncogene point mutations in arthritic synovium.
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DOI：
10.1002/art.1780400913
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发表时间：
1997-09
影响因子：
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通讯作者：
Anne Roivainen;J. Jalava;Laura Pirilä;T. Yli-Jama;Hannu Tiusanen;Paavo Toivanen
中科院分区：
文献类型：
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作者：
Anne Roivainen;J. Jalava;Laura Pirilä;T. Yli-Jama;Hannu Tiusanen;Paavo Toivanen
目的比较类风湿关节炎（RA）患者与骨关节炎（OA）或其他关节病患者滑膜标本中ras原癌基因的突变激活情况。没有任何关节疾病迹象的尸体滑膜样本作为对照材料。方法采用聚合酶链反应（PCR）和PCR扩增产物自动测序相结合的方法，对H-、K-和N-ras原癌基因的密码子12、13和61周围区域进行分析。突变的确认是基于限制性片段长度多态性分析和/或寡核苷酸杂交。结果72例RA患者中有4例（6%），16例OA患者中有2例（13%），12例其他关节病患者中有1例（8%）携带H-ras原癌基因突变，并且在密码子13处GGT- >GAT （Gly- >Asp）突变为杂合。在H-ras基因中发现了一个意想不到的突变，在密码子14上观察到一个杂合的GTG- >ATG （Val- >Met）突变。这种突变的发生率在RA患者中为39%(72 / 28)，在OA患者中为94%(16 / 15)，在其他关节病患者中为42%（12 / 5）。所有携带H-ras密码子13突变的样本同时存在密码子14突变，即存在双突变。在从尸体中获得的一些滑膜标本（n = 8）中也检测到相同的点突变，包括一例双突变。所有标本均显示正常的K-和N-ras位点。结论通过密码子13和14的点突变激活原癌基因H-ras在RA、OA或其他关节病患者的滑膜组织中也存在，在一定程度上也存在于对照滑膜中，表明这种现象并非RA特异性的。在密码子14中，H-ras点突变在OA组织中发生率最高。这个密码子14突变的H-ras基因的可能意义需要澄清。
OBJECTIVE To examine mutational activation of ras proto-oncogenes in synovial tissue from patients with rheumatoid arthritis (RA) compared with synovial specimens from patients with osteoarthritis (OA) or other arthropathies. Synovial samples from cadavers, without any signs of joint disease, were used as control material. METHODS Using a combination of polymerase chain reaction (PCR) and automated sequencing of the amplified PCR product, regions around codons 12, 13, and 61 of the H-, K-, and N-ras proto-oncogenes were analyzed. Confirmation of mutations was based on restriction fragment length polymorphism analysis and/or oligonucleotide hybridization. RESULTS Four (6%) of 72 patients with RA, 2 (13%) of 16 with OA, and 1 (8%) of 12 with other arthropathies harbored mutant H-ras proto-oncogenes, and were heterozygous at codon 13 for the GGT-->GAT (Gly-->Asp) change. An unexpected mutation was found in the H-ras gene, in which a heterozygous GTG-->ATG (Val-->Met) mutation was observed over codon 14. The incidence for this mutation was 39% (28 of 72) in RA patients, 94% (15 of 16) in OA patients, and 42% (5 of 12) in patients with other arthropathies. All samples carrying the codon 13 mutation of H-ras were also codon 14-mutated, i.e., double mutations existed. Identical point mutations were also detected in a few synovial specimens obtained from cadavers (n = 8), including a single case of double mutation. All specimens showed normal K- and N-ras loci. CONCLUSION Activation of proto-oncogene H-ras by point mutation in codons 13 and 14 occurred in the synovial tissue of patients with RA, OA, or other arthropathies, as well as, to some extent, in the control synovia, indicating that the phenomenon is not specific for RA. In codon 14, incidence of the H-ras point mutation was highest in OA tissue. The possible significance of this codon 14-mutated H-ras gene needs to be clarified.