CAJ | 학술논문

器官移植%器官移植基础实验%ABO血型%正反定型不符%ABO基因%双重复合糖基转移酶%启动子%序列测定%多态性%CpG岛%基因甲基化%表观遗传器官移植%器官移植基礎實驗%ABO血型%正反定型不符%ABO基因%雙重複閤糖基轉移酶%啟動子%序列測定%多態性%CpG島%基因甲基化%錶觀遺傳
기관이식%기관이식기출실험%ABO혈형%정반정형불부%ABO기인%쌍중복합당기전이매%계동자%서렬측정%다태성%CpG도%기인갑기화%표관유전
organ transplantation%basic experiments of organ transplantation%ABO blood type%ABO blood grouping discrepancies%ABO gene%glycosyltransferase with dual donor specificity%promoter%sequence analysis%polymorphism%CpG island%gene methylation%epigenetics

　　背景：在ABO血型抗原的研究中，绝大多数样本是相同的ABO基因表达出正常的相同的ABH抗原。但有一定数量的样本表现出具有相同分子遗传背景但表达出来的抗原强度却随着家系\个体不同而有所差异，说明了ABO血型中复杂的表达调控机制。分析一类罕见的双重复合型标本的ABO血型血清学与基因背景情况，深入表观遗传学的研究，有利于部分揭示ABO基因表达机制。目的：探讨双重复合型ABO糖基转移酶表达相关的ABO基因启动子CpG岛甲基化水平与ABH抗原表达的关系。方法：6例经血型血清学定为CisAB或B(A)型的标本，进行ABO基因编码区全长序列和启动子序列的测定，采用重亚硫酸盐处理法检测ABO基因启动子CpG岛甲基化程度。结果与结论：6例双重复合 AB 型的标本中，在 B101等位基因基础上存在 nt803C > G 突变的2个CisAB05/B(A)06等位基因，在ABO启动子CpG岛区域两者在nt-33(30%)、nt+27(50%)、nt+49(50%)具有甲基化差异；在A101等位基因序列的基础上存在nt803C > G突变的2个CisAB01等位基因，在ABO启动子CpG岛区域两者在nt-26(10%)位置有甲基化差异；在B101等位基因基础上存在nt640A > G突变的2个B(A)04等位基因ABO启动子CpG岛，在nt-33(10%)、nt+16(50%)、nt+57(60%)、nt+59(60%)、nt+68(60%)和nt+74(60%)位有甲基化差异。全部的6例标本ABO基因启动子区域DNA序列无任何突变异常。结果提示在相同的ABO遗传基因背景下，ABO基因启动子CpG岛区域某些位点甲基化可能影响ABH抗原在红细胞膜表面的表达。
　　배경：재ABO혈형항원적연구중，절대다수양본시상동적ABO기인표체출정상적상동적ABH항원。단유일정수량적양본표현출구유상동분자유전배경단표체출래적항원강도각수착가계\개체불동이유소차이，설명료ABO혈형중복잡적표체조공궤제。분석일류한견적쌍중복합형표본적ABO혈형혈청학여기인배경정황，심입표관유전학적연구，유리우부분게시ABO기인표체궤제。목적：탐토쌍중복합형ABO당기전이매표체상관적ABO기인계동자CpG도갑기화수평여ABH항원표체적관계。방법：6례경혈형혈청학정위CisAB혹B(A)형적표본，진행ABO기인편마구전장서렬화계동자서렬적측정，채용중아류산염처리법검측ABO기인계동자CpG도갑기화정도。결과여결론：6례쌍중복합 AB 형적표본중，재 B101등위기인기출상존재 nt803C > G 돌변적2개CisAB05/B(A)06등위기인，재ABO계동자CpG도구역량자재nt-33(30%)、nt+27(50%)、nt+49(50%)구유갑기화차이；재A101등위기인서렬적기출상존재nt803C > G돌변적2개CisAB01등위기인，재ABO계동자CpG도구역량자재nt-26(10%)위치유갑기화차이；재B101등위기인기출상존재nt640A > G돌변적2개B(A)04등위기인ABO계동자CpG도，재nt-33(10%)、nt+16(50%)、nt+57(60%)、nt+59(60%)、nt+68(60%)화nt+74(60%)위유갑기화차이。전부적6례표본ABO기인계동자구역DNA서렬무임하돌변이상。결과제시재상동적ABO유전기인배경하，ABO기인계동자CpG도구역모사위점갑기화가능영향ABH항원재홍세포막표면적표체。
BACKGROUND:During the research of ABO blood type antigen, the overwhelming majority samples of same ABO gene express a normal and same ABH antigen. But a certain amount samples with the same ABO genetic background show different antigen intensity expression as for different family or individuals. The ABO blood type has complex expression regulation mechanism. Analysis of ABO blood group serology and genetic background of these rare bi-specific AB phenotype specimens, and further studying on epigenetics may partly revealed ABO gene expression mechanism. OBJECTIVE:To study methylation of CpG island and explore the relationship between ABO gene promoter coding glycosyltransferase with dual donor specificity and ABH antigen expression. METHODS:Six samples detected as CisAB or B(A) phenotype were studied in this paper. The whole code sequences and promoter sequence of ABO gene were amplified respectively. The level of CpG methylation in promoter of ABO gene was further detected with bisulfite treatment method. RESULTS AND CONCLUSION:Among the six bi-specific AB phenotype samples, two previously-identified CisAB05/B(A)06 al eles with nt803C>G on the basis of B101 al ele sequence could be seen, and three additional methylated sites nt-33(30%), nt+27(50%) and nt+49(50%) were found between the two regions of CpG island in promoter of ABO gene. Two CisAB01 al eles with nt803C>G mutation on the basis of A101 sequence were found at nt-26C(10%). Other two B(A)04 al eles contained nt640A>G mutation on the basis of B101 sequence were found in the whole code sequences regions, and six additional methylated sites nt-33(10%), nt+16(50%), nt+57(60%), nt+59(60%), nt+68(60%) and nt+74(60%) were found between the two samples. No abnormity was identified in the promoter region of ABO gene. Our results indicated that the differential methylation levels in the CpG island of ABO gene promoter region may affect ABH antigens expression on the red cel membrane even if the samples had the same ABO genetic background.