慢性粒细胞白血病353例的染色体核型分析

染色体核型分析对于慢性粒细胞白血病 (慢粒 )的诊断、鉴别诊断以及疗效观察具有重要意义。我院 1995年以来收治的 35 3例慢粒的染色体核型资料作一回顾性总结 ,以探讨其细胞遗传学特点和意义。一、资料与方法1 对象 :35 3例慢粒病例为 1995年 1月～ 2 0 0 1年 2月的门诊或住院患者 ,其中男 2 38例 ,女 115例 ,男女比为 2 0 7∶1,年龄 12～ 78岁。临床分期 :慢性期 (ＣＰ) 310例 ,加速期 (ＡＰ) 3例 ,急变期 (ＢＣ) 4 0例 ,后者中有 9例初诊时为慢性期 ,分别于 2个月至 5年后发生急变。慢粒的诊断和分期参照文献 [1]。2 染色体分析 :染色…     

慢性粒细胞白血病经干扰素α治疗Ph~1染色体及M－BCR基因重排消失1例

慢性粒细胞白血病经干扰素α治疗Ｐｈ~１染色体及Ｍ－ＢＣＲ基因重排消失１例南通医学院附属医院葛秀清日本广岛大学原爆放射医学研究所镰田七男关键词白血病．慢性粒细胞性，干扰素，Ｐｈ~１染色体，Ｍ－ＢＣＲ基因报告１例慢性粒细胞白血病（慢粒）以干扰素（ＩＦＮ）α...     

实时定量PCR监测造血干细胞移植后BCR/ABL融合基因的表达

目的监测慢性粒细胞白血病（CML）患者异基因造血干细胞移植（allo-HSCT）后BCR／ABL融合基因表达水平的动态变化,从而判断移植效果,指导临床治疗。方法应用Taqman实时定量RT-PCR方法,以B-actin作为内参照,检测10例初发CML患者及25例接受allo-HSCT治疗的CML患者在移植前、后不同时段外周血中BCR／ABL mRNA表达水平。结果15例接受移植的患者移植前、移植后1、2个月的NBCR／ABL（％）中位数分别为：6．57（0．14～38．83）、0．10（0～1．71）、0（0～0．52）,3个月后全部为0。移植后早期检测BCR／ABL转录本较移植前逐渐下降,移植后1、2个月NBCR／ABL（％）均较移植前下降（x^2均为13．07,P〈O．01）;移植后2个月较1个月下降（x^2=8．10,P〈0．01）。其余10例患者从移植后3～43个月不同时段起连续检测NBCR/ABL（％）也全部为0。结论实时定量RT-PCR方法检测CML患者的BCR／ABL融合基因的表达,操作迅速,灵敏度、特异性好,用于移植后微小残留病的检测对评估预后并指导临床治疗具有重要意义。     

异基因造血干细胞移植治疗慢性粒细胞白血病的疗效及预后因素分析

目的 探讨异基因造血于细胞移植（allo—HSCT）治疗慢性粒细胞白血病（CML）的疗效及预后因素分析。方法选择104例CML患者,采用Bu＋cy、改良Bu＋Cy、TBI＋CY及非清髓方案预处理后行allo—HSCT治疗。结果除1例未植活外,其余均持久性植活。3年无病生存率（DFS）为74．5％,5年累积生存率（OS）为70％。CML慢性期移植、Ⅰ-Ⅱ度急性移植物抗宿主病（GVHD）患者3年DFS分别高于CML加速期／急变期移植、Ⅲ～Ⅳ度急性GVHD患者。多因素Cox回归分析显示,疾病状态、移植类型、急性GVHD的严重程度是异基因HSCT患者长期生存的独立影响因素。结论慢性期且有HLA相合同胞供者的CML患者行allo-HSCT可获得较高长期生存率。     

应用双色荧光原位杂交检测慢性粒细胞白血病衍生9号染色体缺失的研究

目的 研究慢性粒细胞白血病(chronic myelogenous leukemia,CML)患者衍生9号染色体[derivative chromo-some 9,der(9)]部分序列缺失情况,探讨双色荧光原位杂交(FISH)技术检测der(9)部分缺失的应用价值.方法 对2002年3月至2005年12月中国协和医科大学血液学研究所150例CML患者采用不加任何刺激剂的骨髓24 h短期培养法制备染色体,R显带进行核型分析.应用bcr-abl双色双融合DNA探针对骨髓间期细胞行荧光原位杂交,检测der(9)部分序列缺失.结果 124例CML患者进行了染色体核型分析,其中3例分析失败,121例患者中典型Ph染色体易位97例,变异易位24例,同时伴附加染色体异常19例.150例CML患者均进行了双色荧光原位杂交检测,其中27例患者der(9)部分缺失,发生率为18.00%.9例为典型Ph染色体易位,占典型Ph染色体易位患者的9.27%;12例为变异易位,占变异易位患者的50.00%.变异易位患者中的der(9)缺失发生率明显高于典型易位患者,差异有统计学意义(P<0.01).结论 在CML患者中der(9)部分缺失的发生率约为18.00%,应用双色双融合FISH技术能够简便、快速、灵敏的检测出CML患者der(9)缺失.     

Ph~1阴性的慢性粒细胞白血病的重新评价

ph~1是 CGL 的特征性异常。然而,人们发现有些病例虽然具有 CGL 的临床和血液学表现,却缺乏 Ph~1。前者通常称为 Ph~1阳性的 CGL[Ph~1(+)CGL],后者即所谓 Ph~1阴性的 CGL[Ph~1(-)CGL]。这一概念为 Kenis 等首先提出。旧文献中 Ph~(-)CGL 约占全部 CGL病例的10～36％。各家的报道不同,可能与所用的诊断标准不一致有关。近年来,随着深入的研究,人们越来越倾向于认为 Ph~1(-)CGL 是一组异质性疾病,其     

细胞原位RT—PCR检测BCR／ABL融合基因方法的建立与应用

目的：探讨一种新的检测BCR／ABL融合基因方法。方法：在试管内将固定和消化后的细胞直接进行原位逆转录并扩增,反应结束后在显微镜下原位检测。结果：BCR／ABL融合基因表达信号为深紫蓝色或棕色沉淀。36例慢性粒细胞性白血病（CML）阳性检出率达94．4％。其中5例Ph^-CML有3例检出BCR／ABL融合基因表达。急变期和加速期基因表达水平明显高于慢性期病例。结果：细胞原位RT－PCR方法简便快速,灵敏度高,特异性强。不仅可以定性检测BCR／ABL融合基因表达,而且可以量化基因表达水平及其变化。为CML的诊断鉴别诊断以及预测的进展提供了直观和精确的依据。     

Mechanisms of Transformation by the BCR/ABL Oncogene

The Philadelphia chromosome generates a chimeric oncogene in which the BCR and c-ABL genes are fused. The product of this oncogene, BCR/ABL, has elevated ABL tyrosine kinase activity, relocates to the cytoskeleton, and phosphorylates multiple cellular substrates. BCR/ABL transforms hematopoietic cells and exerts a wide variety of biological effects, including reduction in growth factor dependence, enhanced viability, and altered adhesion of chronic myelocytic leukemia (CML) cells. Elevated tyrosine kinase activity of BCR/ABL is critical for activating downstream signal transduction and for all aspects of transformation. This review will describe mechanisms of transformation by the BCR/ABL oncogene and opportunities for clinical intervention with specific signal transduction inhibitors such as STI-571 in CML.     

Apparent decrease and elimination of BCR/ABL mRNA-expressing residual cells in patients with chronic myelogenous leukemia after allogeneic bone marrow transplantation

Summary A modified two-step polymerase chain reaction (PCR) was used for the amplification of BCR/ABL mRNA in 16 patients with Philadelphia chromosomepositive (Ph+) chronic myelogenous leukemia (CML) following allogeneic bone marrow transplantation (BMT). At different intervals after BMT, patient cells were assessed for the presence of BCR/ABL mRNA by two subsequent rounds of PCR amplification; this procedure increased the sensitivity for the detection of one Ph+ cell in 10^4–5 to one cell in 10^5–6. Eight of 16 patients were negative by two-step PCR 1–39 months after BMT, suggesting an elimination of Ph-positive cells or a decrease below the threshold of detection. Although five patients showed negative results by the one-step PCR only, they were tested positive when nested primers were used, indicating a substantial decrease in the amount of BCR/ABL target mRNA compared with earlier pre- or post-transplant analyses. One patient who was still PCR positive 27 months after BMT became negative 12 months later. Persistence of BCR/ABL mRNA-expressing cells correlated with subsequent clinical relapse only when the transplantation was performed during blast crisis. All patients who underwent transplantation in chronic phase, including those with BCR rearrangement by PCR, are in clinical and hematological remission between 24 and 95 months after BMT. We conclude that aggressive chemotherapy combined with total body irradiation is unable to completely eradicate the malignant clone in all CML patients, and it might be speculated that other mechanisms (e.g., graft versus host reaction [GVHD] or graft versus leukemia effect [GVL]) may effectively eliminate residual leukemic cells.The studies were supported by grant Do 176/5-1 from theDeutsche Forschungsgemeinschaft     

Priapism complicating chronic granulocytic leukemia

Since 1952 we have seen nine patients with priapism leading to a diagnosis of chronic granulocytic leukemia (CGL) and a tenth patient who gave a history of priapism when CGL was diagnosed as a result of other symptoms. Seven patients had had one or more transient episodes of prolonged erection before the diagnosis of CGL was established. All ten had high blood leukocyte counts (mean 380 × 10⁹/liter, range 186-782) in comparison with other newly diagnosed patients. We estimate the incidence of this complication at 1%–2% of all male patients presenting with CGL. Treatment of patients in this series varied greatly. Five patients were treated mainly by local measures with or without cytotoxic drugs at conventional dosage, three were treated by sapheno-cavernous bypass operations and leukapheresis followed by cytotoxic drugs at high dosage, and two were treated initially by leukapheresis alone. In general, the prompt initiation of measures designed to reduce the leukocyte count seemed more valuable than the surgical procedures employed in these patients.     

Complex chromosomal abnormalities in a patient with lymphoid blast crisis of chronic myelogenous leukemia

A 46-year-old Chinese man underwent lymphoid blast crisis (Ia+, CALLA+, TdT+) after 5 years of chronic phase, Philadelphia-chromosome positive chronic myelogenous leukemia. Chromosome analysis revealed a hyperdiploid karyotype, including two Philadelphia chromosomes--55,XY,t(9;22) (q34;q11), +2, +5, +5, +6, +10, +18, +19, +21, +del(22)(qll----qter)--in the majority of the leukemic blasts. The constellation of a lymphoid blast crisis and complex chromosomal abnormalities usually associated with myeloid blast crisis as well as the clinical data are discussed.     

Blood outgrowth endothelial cells from chronic myeloid leukaemia patients are BCR/ABL1 negative

The existence of adult haemangioblasts with dual haematopoietic and endothelial developmental potential was confirmed after detection of Ph⁺ vascular endothelial cells in chronic myeloid leukaemia (CML) patients. Blood outgrowth endothelial cells (OECs) from CML patients were found not to harbour the Philadelphia translocation and were thus not clonally related to BRC/ABL1 ⁺ hematopoietic progenitors, but comprised a distinct subfraction of endothelial cells. Remarkably, the frequency of CML-derived OECs was 9-fold higher as compared to healthy donors ( n  = 19 and n  = 300, respectively; P  <   0·0001) and these cells showed increased proliferative potential, possibly reflecting the mobilisation of OEC progenitors by pro-angiogenic cytokines.     

p185(BCR/ABL) has a lower sensitivity than p210(BCR/ABL) to the allosteric inhibitor GNF-2 in Philadelphia chromosome-positive acute lymphatic leukemia

Background

The t(9;22) translocation leads to the formation of the chimeric breakpoint cluster region/c-abl oncogene 1 (BCR/ABL) fusion gene on der22, the Philadelphia chromosome. The p185^BCR/ABL or the p210^BCR/ABL fusion proteins are encoded as a result of the translocation, depending on whether a “minor” or “major” breakpoint occurs, respectively. Both p185^BCR/ABL and p210^BCR/ABL exhibit constitutively activated ABL kinase activity. Through fusion to BCR the ABL kinase in p185^BCR/ABL and p210^BCR/ABL “escapes” the auto-inhibition mechanisms of c-ABL, such as allosteric inhibition. A novel class of compounds including GNF-2 restores allosteric inhibition of the kinase activity and the transformation potential of BCR/ABL. Here we investigated whether there are differences between p185^BCR/ABL and p210^BCR/ABL regarding their sensitivity towards allosteric inhibition by GNF-2 in models of Philadelphia chromosome-positive acute lymphatic leukemia.

Design and Methods

We investigated the anti-proliferative activity of GNF-2 in different Philadelphia chromosome-positive acute lymphatic leukemia models, such as cell lines, patient-derived long-term cultures and factor-dependent lymphatic Ba/F3 cells expressing either p185^BCR/ABL or p210^BCR/ABL and their resistance mutants.

Results

The inhibitory effects of GNF-2 differed constantly between p185^BCR/ABL and p210^BCR/ABL expressing cells. In all three Philadelphia chromosome-positive acute lymphatic leukemia models, p210^BCR/ABL-transformed cells were more sensitive to GNF-2 than were p185BCR/ABL-positive cells. Similar results were obtained for p185^BCR/ABL and the p210^BCR/ABL harboring resistance mutations.

Conclusions

Our data provide the first evidence of a differential response of p185^BCR/ABL- and p210^BCR/ABL- transformed cells to allosteric inhibition by GNF-2, which is of importance for the treatment of patients with Philadelphia chromosome-positive acute lymphatic leukemia.