获得性重型再生障碍性贫血患者穿孔素基因突变检测  

Perforin Gene Mutations in Patients with Acquired Severe Aplastic Anemia

作　　者：张剑[1] 付蓉[1] 王君[1] 李丽娟[1] 宋嘉[1] 瞿文[1] 王化泉[1] 邢莉民[1] 刘鸿[1] 吴玉红[1] 关晶[1] 王国锦[1] 王晓明[1] 梁勇[1] 阮二宝[1] 刘惠[1] 邵宗鸿[1] 

机构地区：[1]天津医科大学总医院血液肿瘤科,天津300052

出　　处：《中国实验血液学杂志》2011年第2期431-434,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金(编号30971286;30971205);天津市应用基础及前沿技术研究计划(编号08jcybjc07800;09jcybjc11200);天津市科技计划项目(编号)07zcgysf00600;高等学校博士学科专项科研基金(编号200800620004)

摘　　要：本研究旨在探讨穿孔素(perforin)基因PRF1突变是否为获得性重型再生障碍性贫血(SAA)发病的遗传易感性基础。用聚合酶链反应(PCR)方法扩增31例SAA患者及15名正常对照者外周血单个核细胞基因组DNAPRF1外显子2(exon2)及外显子3(exon3);用双脱氧终止法测序,与GenBank报道序列核对寻找突变位点;发现突变序列克隆入M13噬菌体载体中,对所得2条染色体相应序列分别测序,以确定不同突变在染色体上的分布。结果表明:在SAA患者,发现了2处基因突变即822位C>T纯合子突变(无义突变)及907位G>A杂合子突变(Met303Val);rs885822单核苷酸多态性(SNP)位点分布病例组与对照组比较有统计学差异(p<0.05)。结论:穿孔素基因突变可能是部分SAA患者的遗传易感因素。This study was aimed to explore whether the perforin gene 1（PRF1） mutation is the basis of genetic susceptibility to pathogenesis of acquired severe aplastic anemia（SAA）.DNA exon2 and exon3 of PRF1 gene in peripheral blood mononuclear cells in 31 SAA patients and 15 normal controls were amplified by PCR;the sequencing was performed by using ABI pRISM 373OXL sequencer;the mutation loci were sought through checking sequences with GenBank-reparted sequences;after the mutation sequences were found,those were cloned into M13 phage vector,then the corresponding sequences of gained 2 chromosomes were sequenced respectively to determine the distribution of different mutations on chromosomes.The results showed that（1） one homozygous mutation（822 CT,synonymous mutation） and one heterozygous mutation（907 GA,methionine 303 valine）were found in PRF1 coding region of 2 SAA patients.These mutations were not detected in normal controls.（2） 1 SNP（rs885822） in the coding region was detected in SAA patients and controls,and the heterozygosity rate between the 2 groups was different（p〈0.05）.It is concluded that perforin gene mutation may be one risk factor in the aberrant proliferation and activation of cytotoxic T cells in pathogenesis of a part of patients with aplastic anemia.

关 键 词：再生障碍性贫血 获得性重型再生障碍性贫血 穿孔素 基因突变 

分 类 号：R556.5[医药卫生—血液循环系统疾病]