CAJ | 학술논문

目的探讨初诊急性髓细胞白血病(acute myeloid leukemia,AML)患者NPM1基因突变发生率及其与染色体核型和FAB亚型之间的关系,并分析NPM1基因的突变类型.方法选取2004至2010年中日友好医院血液科99例初诊AML患者.采集患者骨髓标本,采用聚合酶链反应(polymerase chain reaction,PCR)扩增基因组DNA,并采用变性聚丙烯酰胺凝胶电泳(polyacrylamide gel electrophoresis,PAGE)和毛细管电泳两种方法对AML患者NPM1基因突变进行检测.应用G显带方法对其中72例初诊AML患者进行细胞遗传学分析,同时对10例NPM1突变阳性患者进行直接测序分析.NPM1插入突变在各亚型患者中发生率的比较采用x2检验.变性PAGE和毛细管电泳两种方法检测NPM1基因突变发生率的比较采用McNemar检验.结果毛细管电泳法与变性PAGE法检测AML患者NPM1基因插入突变发生率分别为15％ (15/99)和11％ (11/99),差异无统计学意义(x2 =2.25,P＞0.05).NPM1插入突变在各亚型患者中发生率分别为:急性粒细胞白血病部分分化型(M2)(27％,8/30)、急性单核细胞白血病(M5)(32％,6/19)、红白血病(M6)(13％,1/8),差异无统计学意义(x2=1.06,P＞0.05),其余亚型未检测到NPM1插入突变.49例AML异常核型患者的NPM1插入突变发生率为4％ (2/49),23例正常核型患者的NPM1插入突变发生率为26％ (6/23),差异有统计学意义(x2=5.61,P＜0.05).10例NPM1基因插入突变均为A型突变(c.860_863 dupTCTG).突变导致NPM蛋白羧基末端读码框移,末尾7个氨基酸WQWRKSL被11个氨基酸CLAVEEVSLRK所代替.2例患者检测到内含子缺失突变,分别为IVS10-18_-15delCTTT和IVS10-17-15delTTT.结论 NPM1插入突变为AML患者常见基因改变,正常核型患者插入突变发生率高于异常核型患者.在NPM1基因内含子区发现2例缺失突变.
목적 탐토초진급성수세포백혈병(acute myeloid leukemia,AML)환자NPM1기인돌변발생솔급기여염색체핵형화FAB아형지간적관계,병분석NPM1기인적돌변류형.방법 선취2004지2010년중일우호의원혈액과99례초진AML환자.채집환자골수표본,채용취합매련반응(polymerase chain reaction,PCR)확증기인조DNA,병채용변성취병희선알응효전영(polyacrylamide gel electrophoresis,PAGE)화모세관전영량충방법대AML환자NPM1기인돌변진행검측.응용G현대방법대기중72례초진AML환자진행세포유전학분석,동시대10례NPM1돌변양성환자진행직접측서분석.NPM1삽입돌변재각아형환자중발생솔적비교채용x2검험.변성PAGE화모세관전영량충방법검측NPM1기인돌변발생솔적비교채용McNemar검험.결과 모세관전영법여변성PAGE법검측AML환자NPM1기인삽입돌변발생솔분별위15％ (15/99)화11％ (11/99),차이무통계학의의(x2 =2.25,P＞0.05).NPM1삽입돌변재각아형환자중발생솔분별위:급성립세포백혈병부분분화형(M2)(27％,8/30)、급성단핵세포백혈병(M5)(32％,6/19)、홍백혈병(M6)(13％,1/8),차이무통계학의의(x2=1.06,P＞0.05),기여아형미검측도NPM1삽입돌변.49례AML이상핵형환자적NPM1삽입돌변발생솔위4％ (2/49),23례정상핵형환자적NPM1삽입돌변발생솔위26％ (6/23),차이유통계학의의(x2=5.61,P＜0.05).10례NPM1기인삽입돌변균위A형돌변(c.860_863 dupTCTG).돌변도치NPM단백최기말단독마광이,말미7개안기산WQWRKSL피11개안기산CLAVEEVSLRK소대체.2례환자검측도내함자결실돌변,분별위IVS10-18_-15delCTTT화IVS10-17-15delTTT.결론 NPM1삽입돌변위AML환자상견기인개변,정상핵형환자삽입돌변발생솔고우이상핵형환자.재NPM1기인내함자구발현2례결실돌변.
Objective To analyze the frequency of NPM1 mutation in de novo acute myeloid leukemia (AML) patients and the relationship between NPM1 mutation and chromosome alterations,as well as FAB subgroups,and to analyze the mutation type.Methods A total of 99 de novo AML patients from 2004 to 2010 in China-Japan Friendship Hospital were studied.Genomic DNA was amplified by polymerase chain reaction (PCR),denaturing polyacrylamide gel electrophoresis (PAGE) and capillary electrophoresis were used to detect the mutation of NPM1 gene in 99 AML patients,and karyotyping was performed in 72 AML patients by G banding techniques.DNA sequences analysis of NPM1 mutation was performed on 10 patients.Chi-square test was used to compare the frequencies of NPM1 mutation among the different subgroups,and McNemar's test was used to compare the different rates between denaturing PAGE and capillary electrophoresis.Results The frequencies of NPM1 mutations were detected in 15％ (15/99) of AML patients with capillary electrophoresis and 11％ (11/99 ) with denaturing PAGE(x2 =2.25,P ＞0.05 ).The NPM1 was at different rates in M2(27％,8/30),M5(32％,6/19),M6( 13％,1/8),respectively (x2 =1.06,P ＞ 0.05 ),and not detected in the other subgroups.NPM1 mutation in patients with normal karyotype(26％ ) was more prevalent than patients with abnormal karyotype (4％) (x2 =5.61,P ＜ 0.05)All of the 10 patients were of A type ( c.860_863dupTCTG).The C-terminal portion of the NPM protein by replacing the last seven amino acids(WQWRKSL) with 11 residues (CLAVEEVSLRK).Two intronic deletions were novel,one case was IVS10-18_-15delCTTT,the other was IVS10-17_-15delTTT.Conclusions NPM1 mutations represents a common genetic abnormality in AML patients,and NPM1 mutation in patients with normal karyotype is higher than patients with abnormal karyotype.Two new intronic deletion mutations are identified.