25例慢性粒细胞白血病患者bcr/abl融合基因结果分析

目的探讨逆转录聚合酶链反应(RT—PCR)在检测慢性粒细胞白血病(CML)bcr/abl融合基因方面的应用价值。方法对25例慢性粒细胞白血病患儿血标本提取细胞RNA,用RT—PCR方法检测bcr/abl融合基因。结果 25例慢性粒细胞白血病bcr/abl融合基因分析,阳性24例(96%);阴性1例(4%)。结论 RT—PCR检测bcr/abl融合基因敏感性好,稳定性高,对临床治疗和预后判断具有指导意义。     

骨髓增殖性疾病Bcr／Abl融合基因的表达及其意义

目的：为探讨几种骨髓增殖性疾病bcr／abl 融合基因出现的频率及临床意义．方法：采用逆转录多聚酶键反应（RT－PCR）技术,研究了慢性粒细胞白血病（CML）,真性红细胞增多症（PV）,原发性血小板增多症（ET）,原发性骨髓纤维化（MF）bcr／abl基因的变化．结果：48例慢性期CML,43例呈现bcr／abl基因阳性,占90％,14例加速,急变期CML,6例出现bcr／abl基因,阳性率为43％,与慢性CML相比P＜0.01;10例PV出现1例bcr／abl基因阳性,占10％,6例ET中未发出bcr／abl基因．2例MF中发现1例bcr／abl基因阳性．结论：bcr／abl基因检测对于CML的临床分型有重要意义,bcr／abl阴性的CML预后差,而bcr／abl基因的阳性有助于初诊时的CML急变与急性非淋巴细胞白血病（AML）M2的鉴别和CML与慢性粒单细地白血病（CMML）的鉴别．在其它三种骨髓增殖性疾病中MF可能与CML关系最为密切,而ET关系较远,少数PV可能转化为CML．     

慢性髓性白血病患者β-catenin的表达及其临床意义

目的:比较β-catenin在慢性髓性白血病(CML)各期中的表达情况,并分析与bcr/abl及伊马替尼细胞遗传学疗效的关系,为探讨CML疾病进展机制及寻找新的治疗靶点提供理论依据。方法:采用RT-PCR和Western blotting方法,检测99例CML患者骨髓单个核细胞中β-catenin mRNA和蛋白质水平的表达,分析与bcr/abl的相关性;94例CML患者服用伊马替尼1年后FISH检测bcr/abl融合基因,分析β-catenin与伊马替尼细胞遗传学疗效的关系。结果:CML急变期、加速期患者β-catenin的表达明显增高(P0.01),而慢性期与正常人无明显差别(P0.05);β-catenin与bcr/abl表达水平无明显相关性(r=0.314,P0.05);未达主要细胞遗传学缓解的CML患者β-catenin明显增高(P0.01)。结论:CML疾病进展阶段β-catenin的表达明显升高,β-catenin的表达与bcr/abl无明显相关,而与伊马替尼疗效有关,β-catenin可能参与CML疾病进展机制,可作为新的治疗靶点。     

慢性粒细胞白血病和原发性骨髓纤维化患者bcr/abl融合基因表达及临床意义

目的探讨bcr/abl融合基因的表达水平在慢性粒细胞白血病(CML)和原发性骨髓纤维化(MF)患者的诊断、疗效及预后观察中的意义.方法采用荧光定量逆转录多聚酶链反应(RT-PCR)技术,对18例初、复治CML、7例MF患者的骨髓或外周血单个核细胞进行了bcr/abl融合基因检测,并对其中2例行异基因骨髓移植(allo-BMT)患者进行了短期随防.结果 13例慢性期(CP)CML,12例不同程度表达bcr/abl融合基因,占92.3%,3例加速期(AP),急变期(BC)CML,2例表达bcr/abl融合基因,占66.7%,2例患者allo-BMT后一年内未检出 bcr/abl融合基因,三者之间有非常显著差异P＜0.01;8例初诊患者WBC数及bcr/abl融合基因表达水平明显高于常规治疗组;7例MF患者有1例表达bcr/abl基因.结论 bcr/abl基因是CML发病的分子基础,定量检测bcr/abl融合基因对于CML的诊断、临床分型、疗效观察及预后判断有重要意义;MF患者可能与CML关系密切,对MF患者应进行跟踪观察.     

慢性髓系白血病bcr-abl融合基因的克隆及其在COS-7细胞中的表达

目的克隆慢性髓系白血病(CML)融合基因bcr abl的cDNA序列 ,并在真核细胞中表达。方法以设计的两条引物扩增bcr abl融合位点两侧的cDNA ,测序后克隆至真核表达载体 pcDNA3.1 ,转染COS 7细胞。取常规培养的细胞进行RT PCR鉴定。结果①PCR扩增出490bp大小的cDNA ,其序列测定除第452位的碱基C突变为T外 ,其余序列均正确。②RT PCR法检测到转染细胞系中 ,有bcr ablmRNA的表达。结论成功地克隆了CML融合基因bcr abl融合位点两侧的490bp 序列并表达 ,为研制bcr abl基因疫苗治疗CML奠定了基础。     

66例血液病患者bcr/abl融合基因、Ph染色体结果分析

CML特异性费城染色体（Philadephia,Ph）是由t（9：22）（q34：11）易位形成。9号染色体原癌基因abl（Abelson protooncogene）易位至22号染色体的断裂点簇集区（breakpoint cluster region,bcr）,发生重排,产生bcr／abl融合基因,引起CML的始动突变。融合基因bcr／abl几乎见于所有的慢性粒细胞性自血病、25％-50％的急性B系淋巴细胞性白血病（ALL）和约5％的急性粒细胞性白血病中,本文统计了进行染色体和ber／abl融合基因检查的初诊血液病66例,对其骨髓细胞染色体核型及bcr／abl融合基因进行分析。     

bcr/abl融合基因与慢性粒细胞白血病急变

目的探讨bcr/abl融合基因的表达在CML急变中的意义。方法采用荧光定量逆转录多聚酶链反应(FQ-PCR)技术，对9例CML患者急变前后的血或骨髓进行bcr/abl检测。结果 CML患者从慢性期到急变期往往伴随着bcr/ablmRNA水平的明显升高。结论bcr/abl是CML发病的分子基础，定量检测bcr/abl融合基因对于CML急变的诊断，疗效观察及预后有重要意义。     

白血病bcr/abl融合基因的定量检测及临床意义

目的探讨慢性粒细胞性白血病(ehron ic myeloid leukem ia CML)ber/ab l融合基因检测及意义。方法采用荧光定量PCR(FQ-PCR)检测56例慢性粒细胞性白血病中ber/ab l融合基因的表达。结果对照组bcr/ab l基因表达均为阴性。56例慢性粒细胞性白血病病人bcr/ab l基因表达阳性率为87.5%(49/56),表达值范围3.0×107-7.9×107基因拷贝/L;33例CML患者Ph染色体分析结果中25例为Ph( )/bcr(十),2例为Ph(-)/bcr( ),4例Ph( )/bcr(-),1例Ph(-)/bcr(-);8例CML患者治疗前ber/ab l基因表达均值为(2.60±0.9)×107基因拷贝/L,治疗后为(1.78±0.6)×107基因拷贝/L,两者具有显著性差异(P<0.05=。结论bcr/ab l融合基因表达水平的定量检测及动态观察,对CML的临床诊断、疗效观察和预后具有重要意义。     

实时荧光定量PCR监测治疗前后BCR-ABL水平的变化

目的探讨实时荧光定量PCR(Quantitative Real-time PCR,RQ-PCR)方法在监测慢性粒细胞白血病(chronic myeloid leukemia,CML)患者治疗效果和预后判断的应用价值。方法应用实时荧光定量PCR方法分别检测CML患者采用异基因造血干细胞移植或伊马替尼治疗前、治疗3个月、治疗6个月、治疗1年时BCR-ABL融合基因的表达水平。结果治疗前,CML患者的BCR-ABL水平分布在164%~469%,造血干细胞移植组的患者的BCR-ABL水平降至0.1%~0.2%,伊马替尼组患者随着治疗BCR-ABL水平逐渐下降,在治疗一年后降至0.1%~0.3%。治疗前后患者的BCR-ABL水平差异有统计学意义(P0.05),造血干细胞移植组与伊马替尼组患者的BCR-ABL水平差异不显著(P0.05)。结论使用RQ-PCR方法监测BCR-ABL水平可以作为CML患者评价治疗效果和判断预后的指标。     

骨髓细胞染色体培养法对评价干扰素治疗慢粒白血病疗效的影响

慢性粒细胞白血病(CML)是一种克隆性疾病,绝大多数患者的白血病细胞有固定的细胞遗传学异常,表现为9号染色体和22号染色体长臂交互易位,即出现Ph染色体.Ph染色体在分子水平上形成bcr/abl融合基因[1].Ph染色体可以出现在CML的所有白血病细胞,故作为发病确诊和微小残留病的主要指标.α-干扰素(α-IFN)是目前CML的首选治疗方案,据认为可以达到细胞遗传学缓解即Ph染色体消失.我们观察了1994-1999年期间应用α-干扰素治疗慢性粒细胞白血病前后Ph+染色体的变化,并比较了应用直接法和短期培养法得到Ph的阴转率,并同时检测了8例患者的bcr/abl融合基因.     

Pathogenetic implication of fusion genes in acute promyelocytic leukemia and their diagnostic utility

Acute promyelocytic leukemia (APL) has been recognized as a discrete subset of hematopoietic malignancies constituting approximately 10% of acute myeloid leukemia cases. The hallmark reciprocal chromosomal translocation t(15;17) involving fusion between the retinoic acid receptor (RARα) gene and promyelocytic leukemia (PML) gene is a characteristic feature in APL which consequently results in the emergence of PML-RARα chimeric gene. This gene has been substantiated to be responsible for cellular transformation and is a prime target of all-trans-retinoic acid (ATRA) as well as arsenic-trioxide (ATO) therapy. Since this initial discovery, about 10 diverse translocation partner genes of RARα have been reported that result in variant APL forms strongly suggesting that disruption of RARα underlies its pathogenesis. The nature of the fusion partner has a significant bearing upon disease characteristics including sensitivity to retinoids and ATO and thereby underpins the need for rapid and accurate diagnosis and also demands a highly specific treatment approach. In this article we laid emphasis on the rearrangement of the RARα gene and its different fusion partners resulting in variant forms of APL, their implication in underlying molecular pathogenesis of APL and also the different diagnostic modalities that should be employed for their rapid and accurate diagnosis.     

t(15;17)的变异型插入易位ins(17;15)(q21;q14q22)的细胞遗传学和分子遗传学研究

目的　报道 1例 t(15 ;17)的变异型插入易位 ins(17;15 ) (q2 1;q14 q2 2 )病例及其染色体涂染、逆转录 - PCR的研究结果。方法　骨髓细胞经直接法或 2 4 h培养和外周血单采白血病细胞培养 6天后制备染色体标本 ,以 R显带技术进行核型分析 ;以 15号和 17号整条染色体涂染探针进行染色体涂染 ;以逆转录 -PCR技术检测 PML - RARα和 RARα- PML融合基因的转录本。结果　该患者骨髓细胞和外周血白血病细胞染色体 R显带核型分析结果均提示 15 q-和 17q ;涂染研究证实 17号染色体长臂插入一段 15号染色体来源的染色体片段 ;逆转录 - PCR检出 PML- RARα融合基因短型转录本 ,未检出 RARα- PML 融合基因的转录本 ,符合 ins(17;15 )所致的遗传学改变。结论　染色体涂染和逆转录 - PCR技术是明确急性早幼粒细胞白血病患者涉及 15和 17号染色体插入易位的可靠手段。     

Promyelocyte morphology. Differentiation of acute promyelocytic leukemia from benign myeloid proliferations

Bone marrow aspirates from patients with acute agranulocytosis or a marked left shift in myeloid maturation can mimic acute leukemia, particularly acute hypergranular promyelocytic leukemia. Bone marrow aspirates from 16 cases of apparent acute promyelocytic leukemia, 4 cases of acute agranulocytosis, and 1 case of a marked myeloid left shift were studied for the presence or absence of differentiating features. Normal or reactive promyelocytes were characterized by prominent paranuclear clear Golgi zones, whereas promyelocytes from true leukemic cases all had heavy azurophilic granules dispersed diffusely throughout the cytoplasm. Prominent Golgi zones in promyelocytes were associated only with benign myeloid conditions and were not observed in acute promyelocytic leukemia. The presence of prominent clear Golgi zones in promyelocytes is an important feature assisting in the distinction between leukemic and benign promyelocytes.     

Leukemias resembling acute promyelocytic leukemia, microgranular variant

Acute promyelocytic leukemia (APL) should be distinguished from other subtypes of acute myeloid leukemia (AML) because of the increased risk of disseminated intravascular coagulation (DIC) and its response to arsenic compounds and retinoids. Some cases of AML seem morphologically similar to the microgranular variant of APL (French-American-British [FAB] AML-M3v) but lack the t(15;17). We evaluated 8 cases of APL-like leukemias for subtle morphologic, cytochemical, immunophenotypic, and cytogenetic differences compared with 5 cases of promyelocytic leukemia/retinoic receptor alpha (PML/RARalpha)-positive APL (FAB AML-M3v). We also evaluated both groups for the presence of DIC. No differences among the groups were noted in blast size, chromatin pattern, nuclear morphologic features, intensity of myeloperoxidase staining, or presence of Auer rods. Immunophenotypes were similar; both types of cases lacked CD34 and HLA-DR and were CD13+ and CD33+. Two cases of APL-like leukemias also were CD56+. DIC was present in 2 patients with M3v. Our study shows that there are no definitive morphologic, cytochemical, or immunophenotypic findings that can distinguish these cases from PML/RARalpha-positive APL.