CAJ | 학술논문

目的研究B抗原减弱血液标本的血清学特性和抗原表达减弱的分子机制.方法选择2007年1月至2010年12月于本中心行常规ABO血型检测样本中,B抗原表达减弱的血液样本10份作为研究对象,纳入研究组；计算机随机选择5份正常B表现型的血液样本作为对照组.采用经典血清学技术检测红细胞ABO血型,聚合酶链反应(PCR)技术扩增ABO基因全部外显子序列和5’端调控(CBF/NF-Y增强子)序列,并进行测序分析,应用逆转录-PCR(RT-PCR)技术分析ABO cDNA转录剪接情况,采用重亚硫酸盐转化和克隆测序法分析ABO基因启动子区CpG岛甲基化水平.结果研究组标本的血清学表现为B抗原明显减弱,全部外显子编码序列和拼接位点碱基无任何突变.ABO基因增强子未发现异常,存在着ABO基因全长cDNA转录本,未发现新的转录剪接体.与对照组比较,研究组10份标本在启动子区CpG岛均呈现不同程度的甲基化,其中6个标本在nt-26序列位点表现半甲基化状态.结论 ABO基因启动子区CpG岛中的甲基化位点可能是引起B抗原弱表达的原因.
목적 연구B항원감약혈액표본적혈청학특성화항원표체감약적분자궤제.방법 선택2007년1월지2010년12월우본중심행상규ABO혈형검측양본중,B항원표체감약적혈액양본10빈작위연구대상,납입연구조；계산궤수궤선택5빈정상B표현형적혈액양본작위대조조.채용경전혈청학기술검측홍세포ABO혈형,취합매련반응(PCR)기술확증ABO기인전부외현자서렬화5’단조공(CBF/NF-Y증강자)서렬,병진행측서분석,응용역전록-PCR(RT-PCR)기술분석ABO cDNA전록전접정황,채용중아류산염전화화극륭측서법분석ABO기인계동자구CpG도갑기화수평.결과 연구조표본적혈청학표현위B항원명현감약,전부외현자편마서렬화병접위점감기무임하돌변.ABO기인증강자미발현이상,존재착ABO기인전장cDNA전록본,미발현신적전록전접체.여대조조비교,연구조10빈표본재계동자구CpG도균정현불동정도적갑기화,기중6개표본재nt-26서렬위점표현반갑기화상태.결론 ABO기인계동자구CpG도중적갑기화위점가능시인기B항원약표체적원인.
Objective To study the serological characteristics and molecular mechanism of blood samples with weaken B antigen.Methods From January 2007 to December 2010,a total of 10 blood samples with weak B antigen which were chosen from conventional ABO blood samples tested blood samples,were included in this study as study group.And a total of 5 blood samples with normal B antigen expression were selected as control group by computer randomly.All the samples were detected ABO blood group by standard serum technique.The whole code sequences and 5' regulatory region containing the CBF/NF-Y enhancer of ABO gene were amplified by polymerase chain reaction (PCR) respectively and direct sequencing.The alternative splicing isoforms of ABO cDNA were analyzed by reverse transcription-PCR (RT-PCR).The level of methylation of the CpG island in ABO gene promoter was further analyzed by bisulfite and cloning sequencing method.Results The serological characteristic of these samples showed that B antigen was obviously decreased.There was no mutation in the full-length coding sequences and splice receptor sites.The nucleotide characteristics of the 5'-UTR was consistent with common allele type and none abnormity was identified in the promoter,enhancer and regulatory sequence regions.The integrative cDNA transcript of ABO gene was obtained and no new splicing isoform was found.Compared with the normal B phenotype samples,10 samples of study group all showed different methylation levels on the promoter CpG sites of ABO gene and nt-26 residues of 6 samples were partially methylated.Conclusions The methylation in the CpG island of ABO gene promoter region may cause weak expression of the B antigen.