中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2008年
7期
459-463
,共5页
陈小文%岳丽杰%李成荣%李长钢%石红松%张民
陳小文%嶽麗傑%李成榮%李長鋼%石紅鬆%張民
진소문%악려걸%리성영%리장강%석홍송%장민
基因,CDA%多态性,单核苷酸%阿糖胞苷%白血病%酵母表达系统
基因,CDA%多態性,單覈苷痠%阿糖胞苷%白血病%酵母錶達繫統
기인,CDA%다태성,단핵감산%아당포감%백혈병%효모표체계통
Gene,CDA%Polymorphism,single nucleotide%Cytarabine%Leukemia%Yeast expression system
目的 探讨人类胞苷脱氨酶(CDA)基因编码区单核苷酸多态性(cSNP)与儿童急性白血病(AL)患者阿糖胞苷(Ara-C)敏感性的关系.方法 采集87例AL患儿骨髓及199例对照儿童的外周血,制备cDNA,采用变性梯度凝胶电泳及DNA测序进行CDA基因cSNP分析.将人类CDA基因分别转入大肠杆菌和酵母细胞,利用高效液相色谱分析确定不同等位型CDA的催化活性;应用生长抑制实验检测酵母转化体对Ara-C处理的敏感性.结果 在对286例中国儿童进行CDA基因cSNP检测中发现了3个已知的cSNP,即79A/C(K27Q)、208G/A(A70T)和435T/C,其等位基因频率分别为12.1%、0.5%和76.2%.这些多态性与白血病的易感性无相关性.携带人类CDA70T的大肠杆菌转化体的胞苷及Ara-C脱氨活性分别较CDA70A降低53%和63%(P值均＜0.01),Ara-C对携带人类CDA70T的酵母转化体作用15 h的细胞半数抑制浓度(IC50)值[(735±31)μmol/L]也显著低于Wt转化体[(973±61)μmol/L](P＜0.05),降低25%.结论 确定了中国人CDA基因中3种cSNP以及各自的基因型分布和等位基因频率.证实了一个人类CDA208A基因型,其对Ara-C的敏感性高于CDA208G基因型,提供了一个对脱氧胞苷同系物潜在敏感的亚群.
目的 探討人類胞苷脫氨酶(CDA)基因編碼區單覈苷痠多態性(cSNP)與兒童急性白血病(AL)患者阿糖胞苷(Ara-C)敏感性的關繫.方法 採集87例AL患兒骨髓及199例對照兒童的外週血,製備cDNA,採用變性梯度凝膠電泳及DNA測序進行CDA基因cSNP分析.將人類CDA基因分彆轉入大腸桿菌和酵母細胞,利用高效液相色譜分析確定不同等位型CDA的催化活性;應用生長抑製實驗檢測酵母轉化體對Ara-C處理的敏感性.結果 在對286例中國兒童進行CDA基因cSNP檢測中髮現瞭3箇已知的cSNP,即79A/C(K27Q)、208G/A(A70T)和435T/C,其等位基因頻率分彆為12.1%、0.5%和76.2%.這些多態性與白血病的易感性無相關性.攜帶人類CDA70T的大腸桿菌轉化體的胞苷及Ara-C脫氨活性分彆較CDA70A降低53%和63%(P值均＜0.01),Ara-C對攜帶人類CDA70T的酵母轉化體作用15 h的細胞半數抑製濃度(IC50)值[(735±31)μmol/L]也顯著低于Wt轉化體[(973±61)μmol/L](P＜0.05),降低25%.結論 確定瞭中國人CDA基因中3種cSNP以及各自的基因型分佈和等位基因頻率.證實瞭一箇人類CDA208A基因型,其對Ara-C的敏感性高于CDA208G基因型,提供瞭一箇對脫氧胞苷同繫物潛在敏感的亞群.
목적 탐토인류포감탈안매(CDA)기인편마구단핵감산다태성(cSNP)여인동급성백혈병(AL)환자아당포감(Ara-C)민감성적관계.방법 채집87례AL환인골수급199례대조인동적외주혈,제비cDNA,채용변성제도응효전영급DNA측서진행CDA기인cSNP분석.장인류CDA기인분별전입대장간균화효모세포,이용고효액상색보분석학정불동등위형CDA적최화활성;응용생장억제실험검측효모전화체대Ara-C처리적민감성.결과 재대286례중국인동진행CDA기인cSNP검측중발현료3개이지적cSNP,즉79A/C(K27Q)、208G/A(A70T)화435T/C,기등위기인빈솔분별위12.1%、0.5%화76.2%.저사다태성여백혈병적역감성무상관성.휴대인류CDA70T적대장간균전화체적포감급Ara-C탈안활성분별교CDA70A강저53%화63%(P치균＜0.01),Ara-C대휴대인류CDA70T적효모전화체작용15 h적세포반수억제농도(IC50)치[(735±31)μmol/L]야현저저우Wt전화체[(973±61)μmol/L](P＜0.05),강저25%.결론 학정료중국인CDA기인중3충cSNP이급각자적기인형분포화등위기인빈솔.증실료일개인류CDA208A기인형,기대Ara-C적민감성고우CDA208G기인형,제공료일개대탈양포감동계물잠재민감적아군.
Objective To study the relationship between coding single-nucleotide polymorphisms(cSNPs)in the human cytidine deaminase(CDA)gene and cytosine arabinoside(Ara-C)sensitivity in childhood acute leukemia(AL).Methods cDNAs from 87 leukemia and 199 control blood samples were analyzed for the cSNPs in CDA by PCR-denaturing gradient gel electrophoresis(DGGE)and sequencing.Human CDA genes were transformed into E.coli and yeast,respectively.Catalytic activities of the allele CDA and variant CDAs were determined by HPLC assay.The Ara-C sensitivity of the yeast transformants Was messured by growth inhibition assays.Results Three known different polymorphisms,namely,79A/C(K27Q),208G/A(ATOT)and 435T/C(silent)were identified in the coding msion of CDA from an investigated Chinese population and displayed allelie frequencies of 12.1%,0.5%and 76.2%,respectively.No association with susceptibility to disease Was observed.Compared with that of CDA70A,the deamination activities for cytidine and Am-C substrates of the E.coli transformants carrying human CDA70T were decreased by 53%and 63%,respectively(P＜0.01),and the Ara-C IC50 value of the yeast transformants was also significantly decreased by 25%[(973±61)μmol/L to(735±31)μmol/L,P＜0.05].Conclusions The 3 known cSNPs and their allelic frequencies of CDA are identified in a Chinese childhood AL.The 208A genotype is shown to be more sensitive to Ara-C than 208G genotype.
