慢性粒细胞白血病的分子生物学特征与临床研究进展

Ｐｈ染色体是慢性粒细胞白血病（ＣＭＬ）的细胞遗传学标记，而ｂｃｒ／ａｂｌ基因重排则为ＣＭＬ的分子基础。ＢＣＲ、ｃ－ａｂｌ基因重排及ｂｃｒ／ａｂｌ嵌合基因的结构，转录和表达是ＣＭＬ寻病的关键环节。随着分子生物学技术的迅速发展，对ｂｃｒ／ａｂｌ重组基因的研究日益深入有可能最终阐明ＣＭＬ的发病机制，并在临床上对ＣＭＬ诊断、分型、鉴别、预后判断及治疗等均有极其重要的指导意义。目前已在尝试利用分子生物学技术     

慢性粒细胞白血病的分子生物学特征与临床研究进展

Ｐｈ染色体是慢性粒细胞白血病（ＣＭＬ）的细胞遗传学标记，而ｂｃｒ／ａｂｌ基因重排则为ＣＭＬ的分子基础。ＢＣＲ、ｃ－ａｂｌ基因重排及ｂｃｒ／ａｂｌ嵌合基因的结构，转录和表达是ＣＭＬ发病的关键环节．随着分子生物学技术的迅猛发展，对ｂｃｒ／ａｂｌ重组基因的研究日益深入有可能最终阐明ＣＭＬ的发病机制，并在临床上对ＣＭＬ诊断、分型、鉴别、预后判断及治疗等均有极其重要的指导意义．目前已在尝试利用分子生物学技术在基因水平根治ＣＭＬ，这项技术为治疗ＣＭＬ开辟了一条新途径。     

慢性粒细胞白血病急性变p53基因改变的初步探讨

慢性粒细胞白血病急性变ｐ５３基因改变的初步探讨陈敬春，刘树茂，费洪宝，龚维龙Ｐｈ染色体易位所致的ｂｃｒ／ａｂｌ融合基因的形成及活化是慢性粒细胞白血病（简称慢位，ＣＭＬ））发病的关键。ＣＭＬ在临床上要经过慢性期（ＣＰ）、加速期（ＡＰ）及急变期（ＢＣ），...     

双色荧光原位杂交检测慢性粒细胞白血病的bcr/abl融合基因

自１９８４年发现了慢性粒细胞白血病（ｃｈｒｏｎｉｃｍｙｅｌｏｉｄｌｅｕｋｅｍｉａ,ＣＭＬ）具有ｂｃｒ／ａｂｌ融合基因［１,２］,检测ｂｃｒ／ａｂｌ融合基因就成为诊断ＣＭＬ的关键。Ｔｋａｃｈｕｋ等［３］使用间接标记的荧光原位杂交（ｆｌｕｏ－ｒｅｓｃｅｎ...     

α—2b干扰素联合低剂量阿糖胞苷治疗慢性粒细胞白血病

α－２ｂ干扰素（ＩＦＮα－２ｂ）与低剂量阿糖胞苷（ＬＤ Ａｒａ－Ｃ）联合应用具有相加作用,可以减少或消除Ｐｈ染色体阳性细胞和ｂｃｒ／ａｂｌ融合基因,显提高慢性粒细胞白血病（ＣＭＬ）患的血液学和细胞遗传学缓解率,延长ＣＭＬ患的生存期,被认为是目前治疗ＣＭＬ最有前途的方案之一。     

bcr基因重排与白血病

Ｐｈ染色体为性粒细胞白血病的特异标记染色体，存在于９５％的ＣＭＬ患者中，近年来随着分子生物学的进展。已阐明Ｐｈ染色体所导致的ｂｃｒ／ａｂｌ基因重排是某些白血病发病的关键因素，本文复习了国内外有关文献，对该领域目前研究的进展做一简要综述。     

慢性粒细胞性白血病急变的相关分子研究进展

慢性粒细胞性白血病（ＣＭＬ）是由ｔ（９，２２）（ｑ３４，ｑ１１）易位引起的一种血液系统恶性疾病。该易位重排形成的ｂｃｒ／ａｂｌ融合基因及其所编码的Ｐ２１０蛋白在ＣＭＬ发生中的作用已被确认。近年研究表明ｂｃｒ／ａｂｌ融合基因、抑瘤基因（如ｐ５３，Ｒｂ，ｐ１６等）及其它分子（如ＥＶＩ１，ｃｍｙｃ等）均与ＣＭＬ急变有关。分析这些相关分子对ＣＭＬ急变的直接或间接影响，对ＣＭＬ的分子病理学及其治疗研究均具指导意义。     

bcr/abl及c-myb原癌基因反义寡核苷酸对K562细胞的作用研究

目的 为反义基因治疗慢粒白血病（ＣＭＬ）奠定实验基础,并进一步提示ＣＭＬ的发病机制。方法 设计并合成针对ｂｃｒ／ａｂｌ嵌合基因和ｃ－ｍｙｂ原癌基因的反义寡脱氧核苷酸（ａｓＯＤＮ）及其硫代衍生物,观察其对Ｋ５６２细胞的生长、ＤＮＡ合成及ｂｃｒ／ａｂｌ基因转录、表达以及细胞凋亡的影响。结果 针对ｂｃｒ／ａｂｌ基因及ｃ－ｍｙｂ癌基因的ａｃＯＤＮ可显著抑制Ｋ５６２细胞的生长、ＤＮＡ合成及ｂｃｒ／ａｂｌ基因     

Bcr/abl嵌合基因表达与慢粒白血病细胞积累

目的和方法 :ｂｃｒ/ａｂｌ嵌合基因是定位于人 9号染色体 9ｑ34上的ｃ -ａｂｌ基因和 2 2号染色体 2 2ｑ1 1上的ｂｃｒ基因因ｔ( 9:2 2 )易位 ,使相应断裂的基因发生融合所形成。它是ｐｈ染色体的分子基础 ,在慢粒白血病 (ＣＭＬ)发病中起重要作用。Ｂｃｒ/ａｂｌ嵌合基因编码具酪氨酸激酶活性的蛋白 (Ｐ2 1 0 )。这种蛋白与细胞信号传导蛋白相互作用 ,扰乱细胞信息传递过程 ,使ＣＭＬ细胞增殖和分化失控。为了探讨ｂｃｒ/ａｂｌ嵌合基因表达致慢粒白血病细胞积累的发病机制 ,设计并合成针对ｂｃｒ/ａｂｌ嵌合基因的反义寡脱氧核苷酸 (ａ …     

骨髓细胞染色体培养法对评价干扰素治疗慢性白血病疗效的影响

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 2 2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…     

Megakaryocytes carry the fused bcr-abl gene in chronic myeloid leukaemia: a fluorescence in situ hybridization analysis from bone marrow biopsies

Histological examination of bone marrow biopsies shows that about one-third of chronic myeloid leukaemia (CML) patients exhibit an increase of megakaryocytes. The megakaryocytic predominance may be so striking that differentiation from other chronic myeloproliferative disorders (CMPD) may be difficult in some CML patients. Megakaryocytes in CML are clonal as demonstrated by loss of glucose-6-phosphate dehydrogenase isoenzymes. The Ph translocation, fusing the abl and bcr genes on chromosomes 9 and 22, however, obviously occurs as a second step in tumour development. So far, the Ph translocation has not been assigned explicitly to megakaryocytes. The question is whether the megakaryocytic cell lineage could harbour the bcr/abl fusion in those CML cases with striking proliferation of megakaryocytes but lack this genetic defect in cases with normal or decreased megakaryocyte counts. We therefore performed triple-colour fluorescence in situ hybridization (FISH) for portions of the bcr and abl genes flanking the breakpoint in CML in paraffin sections of CML cases with normal and with increased numbers of megakaryocytes. This method allows identification of the bcr/abl fusion in single, morphologically intact cells, whereas conventional cytogenetics requires lysis and thus destruction of the cell. Among the 21 CML patients examined by FISH, 10 were informative for bcr and abl genes and displayed distinct hybridization signals within nuclei of bone marrow cells. Besides the granulopoietic cells, megakaryocytes of all those patients (4 without and 6 with varying grades of megakaryocytic increase) displayed bcr/abl fusion signals indiciative of a Ph translocation. The lack of hybridization signals in the remaining 11 cases indicates that this technique is not of value diagnostically and should be reserved for scientific questions. Positive controls consisted of conventional chromosome preparations from bone marrow aspirates demonstrating the Ph chromosome in all patients examined, and negative controls of paraffin sections of bone marrow biopsies from non-CML patients. These showed no fusion signals in bone marrow cells, including megakaryocytes, using FISH. Our results demonstrate clearly that not only the transforming event but also the Ph translocation leading to the bcr/abl fusion happens prior to the differentiation of the pluripotent stem cell into different myeloid lineages. The megakaryocytic proliferation evident in some CML cases is probably a consequence of the disease progress.     

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.     

Clinical implications of fluorescence in situ hybridization analysis in 13 chronic myeloid leukemia cases: Ph-negative and variant Ph-positive.

Thirteen chronic myeloid leukemia (CML) patients, 10 with variant Philadelphia (Ph) translocations and 3 Ph negative cases, were analyzed by fluorescence in situ hybridization (FISH) with the use of BCR and ABL cosmid probes and a chromosome 22 painting probe. In the variant Ph translocations, the BCR-ABL fusion gene was located on the Ph chromosome; in 1 CML Ph-negative patient, the BCR-ABL fusion gene was located on the Ph chromosome; and, in 2 patients, it was located on chromosome 9. The chromosome 22 painting probe was detected on the third-party chromosome of the variant translocation, and in none of the variant translocations was there any detectable signal on chromosome 9. In CML patients with clonal evolution of a simple Ph, a signal of the chromosome 22 painting probe was detected on the der(9) of the Ph translocation. It was concluded that the variant Ph translocations evolved simultaneously in a three-way rearrangement. The clinical parameters of the 13 patients were similar to those of a large group of CML patients with a simple Ph translocation. It is suggested that, to determine the prognosis of CML patients with a complex karyotype, FISH analysis with a chromosome 22 painting probe be performed.     

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．     

Fluorescent in situ hybridization diagnosis of extramedullary nodal blast crisis

The t(9;22)(q34;q11) translocation between bcr and abl genes plays a pivotal role in the pathogenesis and diagnosis of chronic myelogenous leukemia (CML). Fluorescence in situ hybridization (FISH) using specific DNA probes provides a useful and accurate way for the detection of bcr/abl fusion gene in single cell. Here, we report an unusual case of a patient with no prior hematologic disease who initially manifested lymphadenopathy. The lymph node findings were suspicious for T‐lineage lymphoblastic lymphoma, however, his blood and bone marrow at that time were in chronic phase of CML. This presented difficulty for accurate discrimination between CML blast crisis (BC) and non‐Hodgkin's lymphomas (NHLs). To discern where the extramedullary nodal malignancy originated from, we cytologically analyzed lymph node biopsies and bone marrow with FISH to detect bcr/abl fusion signals. Together with the morphology, immunohistochemistry, cytogenetics as well as molecular analysis, the patient was diagnosed as extramedullary T‐lymphoid BC of Ph+ CML. In conclusion, this case is unusual at three levels: first, extramedullary nodal BC as a presenting manifestation of CML is rare and the blasts are of precursor T lymphoblastic lineage, rather than the more common B‐cell lineage; second, this case suggests that extramedullary lymphoid nodal BC of CML can exist independently without the bone marrow developing into BC; and third, FISH analysis on the single neoplastic cell is an accurate way to confirm that the neoplasm is either extramedullary localized blasts of CML or genetically distinct neoplasm. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

Chromosomal in situ hybridization and Southern blot analyses using c-abl, c-sis, or bcr probe in chronic myelogenous leukemia cells with variant Philadelphia translocations

The Philadelphia (Ph) chromosome is a cytogenetic hallmark of chronic myelogenous leukemia (CML). Whereas the majority of Ph-positive CML patients show the standard Ph translocation involving chromosomes 9 and 22, t(9;22)(q34;q11), the minority of cases exhibit a variant type of Ph translocation involving these two and other chromosomes (complex type) or those involving #22 and chromosomes other than #9 (simple type). To get an insight into the nature of variant Ph translocations and the process of their formation, we examined the localization of the c-abl and c-sis oncogenes and the breakpoint cluster region (bcr) gene by chromosomal in situ hybridization in ten variant Ph translocations of CML including five simple and five complex ones as initially interpreted. In situ hybridization showed that c-abl localized to band 9q34 and c-sis localized to band 22q12-q13 were translocated on the Ph and on one of the rearranged chromosomes other than #9, respectively, in all the variant translocations examined. On the other hand, bcr localized to band 22q11 was translocated on various chromosomes but mostly on chromosome 9. Parallel Southern blot analyses on DNA from leukemic cells of five patients including two with simple translocations and three with complex ones revealed rearrangements of bcr with breakpoints occurring mostly in a 5' portion of 5.8-kb BamHI/BglII sequences, which are quite similar to those detected so far in CML cases with the standard Ph translocation. The present findings strongly suggest that variant Ph translocations of CML are all complex, and some of them are formed stepwisely from the standard translocation.     

Interphase fluorescence in situ hybridization studies for the detection of 9q34 deletions in chronic myelogenous leukemia: a practical approach to clinical diagnosis

Chronic myelogenous leukemia (CML) is characterized by the Philadelphia chromosome (Ph) in more than 90% of cases. Recent studies using fluorescence in situ hybridization (FISH) have shown that in a subset of patients with CML, deletions of 9q34 involving the argininosuccinate synthetase region occur at the time of the Philadelphia translocation and are associated with a poor prognosis. We performed interphase FISH studies in 152 cases of CML using a dual-color, dual-fusion probe system with a third probe directed at 9q34. Cytogenetic studies showed a simple (typical) Ph in 124/152 (82%), a cryptic Ph in 11/152 (7%), and a variant Ph chromosome with a complex translocation in 17/152 (11%) of cases. Interphase FISH studies showed single BCR/ABL fusion patterns in 48/152 (32%) of cases. Deletions of 9q34 were observed in 14% of all the cases and were present in 46% of cases with single BCR/ABL fusion pattern. All the 9q34 deletions occurred in cases with single BCR/ABL fusion signal. However, a single-fusion pattern is not specific for 9q34 deletions, and cases should be routinely screened for the presence of this prognostically significant abnormality by using a third probe directed specifically at 9q34.     

荧光原位杂交技术在诊断变异Ph易位及Ph(一)慢性髓细胞白血病中的应用

目的 探讨荧光原位杂交(fluorescence in situ hybridization,FISH)技术在诊断变异Ph易位及Ph(-)慢性髓细胞白血病(chronic myelocytic leukemia,CML)中的应用价值.方法 应用常规R显带方法,对9例伴有变异Ph易位和2例Ph(-)CML患者采用双色双融合bcr/abl探针进行FISH检测.结果 9例变异Ph易位CML患者异常核型除涉及9和22号染色体外,还涉及1、3、5、12、13、15、17、21号染色体,且部分类型为重现性异常,FISH结果均为阳性,信号特征为2R2G1Y;2例Ph(-)CML患者核型正常,FISH结果阳性,信号特征分别为1R1G2Y和1R1G1Y.结论 FISH技术对伴有变异Ph易位及Ph(-)CML患者的诊断更具有优势,可根据阳性细胞信号特征分析其异常核型,判断标记基因异常改变情况,是常规染色体显带分析的有益补充.