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:运用特殊位点(LSI)9q34探针荧光原位杂交技术(FISH)研究慢性粒细胞白血病(CML)患者衍生9号染色体部分序列缺失的情况,并探讨临床意义。方法:对42例CML患者进行常规染色体核型分析,采用BCR/ABL双色双融合或ES探针对骨髓间期细胞进行FISH检测,同时运用9q34探针检测有无衍生9号染色体部分序列缺失。结果:42例染色体核型分析中,35例为典型的t(9;22)染色体易位,4例为复杂易位。42例患者均行FISH检测证实为BCR/ABL融合基因阳性。运用9q34探针检测所有42例患者中发现有6例伴有衍生der(9)部分序列缺失,发生率为14%,发生复杂易位的4例患者均未发现der(9)部分缺失。6例发生缺失的患者仅有1例处于慢性期,其中3例患者病程仅为1-2个月,初诊就已诊断为CML加速期或急变期。结论:在CML患者中der(9)部分缺失的发生率为14%,伴有der(9)部分缺失的患者多表现为慢性期短,建议所有BCR/ABL融合基因阳性的CML患者应常规使用9q34探针检测der(9)有无缺失。     

衍生9号染色体缺失对慢性髓细胞白血病的预后价值

目的研究慢性髓细胞白血病（CML）中衍生9号染色体[der（9）]缺失的发生率及预后意义。方法随机选取48例CML-急变期（BC）患者,以BCR-ABL双色双融合荧光原位杂交（FISH）技术检测其染色体标本。对于FISH技术检测到的间期细胞中只有单个融合信号的标本,则观察其中期细胞,以明确是否为der（9）缺失。对于der（9）缺失患者,以相同方法检测其初诊时的染色体标本,以明确der（9）缺失是否和Ph易位同时发生。患者慢性期以羟基脲治疗。结果48例CML-BC患者中有8例（16．7％）der（9）缺失,其初诊时的染色体标本der（9）也缺失。der（9）缺失组慢性期及总生存期分别平均为20．9和36．4个月,明显短于未缺失组（55．1和70．3个月）（P〈0．05）。CML-BC急淋变与急非淋变患者中der（9）缺失概率无统计学意义（P〉0．05）。结论FISH技术可有效检测der（9）缺失。der（9）缺失与Ph易位同时发生。具有der（9）缺失的CML疾病进展较快,预后较差。羟基脲不具有逆转CML中der（9）缺失的不良预后的作用。der（9）缺失不导致cML向某一特定类型转化。     

荧光原位杂交技术研究慢性粒细胞白血病隐匿型变异易位

目的:探讨荧光原位杂交(FISH)技术对确定慢性粒细胞白血病(CML)隐匿型变异易位的重要作用。方法:骨髓细胞短期培养,常规显带用G显带技术,FISH检测以染色体全长涂抹探针进行。结果:485例CML患者中4例经常规G显带技术未见典型Ph染色体,FISH检出具有隐匿型Ph染色体(0．9％)。结论:FISH技术较传统显带技术准确、快速、特异、敏感,能准确辨认变异易位的类型,给予准确的分子细胞遗传学诊断,在CML的诊断和疗效监测中具有重要意义。     

双色双融合间期荧光原位杂交探针检测慢性髓系白血病的微小残留病状态

目的 研究双色双融合探针间期荧光原位杂交技术（D-FISH）检测慢性髓系白血病（CML）微小残留病（MRD）的灵敏度,特异性及可重复性。方法 应用D-FISH探针对8例移植后或经供者淋巴细胞输注治疗获完全细胞遗传学缓解的CML患者骨髓内残留的微小肿瘤负荷水平进行检测,与应用单融合探针（S-FISH）进行了比较,同时就D-FISH与逆转录-聚合酶链反应（RT-PCR）结果的相关性进行了探讨。结果 D-FISH较S-FISH具有更高的灵敏度及特异性,可重复性好,正常对照组中D-FISH的正常分界值为0.570%,而S-FISH为5.686%。差异具有显著性;对8例移植后患者进行MRD检测S-FISH检测到3/8（37.5%)阳性,而D-FISH检测到5/8例（62.5%)阳性且结果与RT-PCR结果高度相关。结论 由于D-FISH具有较低的假阳性率与假阴性率,可应用于CML治疗后MRD的检测,但检测结果与临床参数的相关性有待进一步探讨。     

应用间期荧光原位杂交技术评估干扰素治疗慢性粒细胞白血病的疗效

目的　探讨间期荧光原位杂交技术 (FISH)检测慢性粒细胞白血病 (CML)干扰素 (IFN)治疗后体内残留Ph阳性细胞的可行性。方法　应用间期FISH对 7例未治疗CML患者和 17例IFN α 2b长期治疗CML患者检测体内Ph阳性细胞变化情况。结果　正常对照组假阳性率为 ( 3 87± 0 94 ) % ,确定正常值为小于 6 68% (x± 3s)。未治疗组Ph阳性细胞平均为 89 2 1%。干扰素治疗组Ph阳性细胞平均为 4 4 86% ,与未治疗组相比无显著性差异 ( P >0 0 5 )。细胞遗传学有效患者 (CGR 4例 ,PGR 2例 ,共 8个标本 )Ph阳性细胞平均为 2 6 3 0 % ,与未治疗组相比有显著性差异 (P <0 0 5 )。而且Ph阳性细胞的减少与INF用药总剂量具有相关性 (r =-0 666,P =0 0 0 2 )。结论　间期FISH检测比染色体核型、RT PCR技术更能准确地评价干扰素治疗后体内存有的白血病细胞的负荷量 ,评价干扰素治疗CML的疗效     

应用荧光原位杂交检测初治慢性淋巴细胞白血病患者的分子遗传学异常

目的探讨荧光原位杂交(FISH)技术检测慢性淋巴细胞白血病(CLL)患者分子遗传学异常的临床意义。方法应用FISH技术,采用4种特异性DNA探针检测88例初治CLL患者的分子遗传学异常,并结合患者临床资料,分析各分子遗传学异常与临床分期的关系。结果 88例CLL患者中,52例出现分子遗传学异常,总异常率为59.1%。RB1基因异常发生率最高,为30.7%,其后依次为CSP12(27.3%)、P53及ATM缺失发生率均约为8.0%。结论 FISH技术检测分子遗传学异常的敏感性和特异性较高。RB1基因异常是CLL最常见的分子遗传学异常。伴P53及ATM基因异常的的CLL分期较晚,预后较差。     

慢性粒细胞白血病BCR/ABL融合基因及复杂变异的染色体核型分析

目的探讨慢性粒细胞白血病(CML)患者BCR/ABL融合基因及其复杂变异的染色体核型变化及临床意义。方法在常规细胞遗传学(CC)方法检测基础上,运用分子生物学方法——荧光原位杂交(FISH)技术,采用多种位点特异性DNA探针(染色体全染、特殊位点、双色易位融合探针),对2002年9月至2007年2月中国医科大学附属第一临床学院血液科56例门诊及住院CML患者(慢性期51例,加速及急变期5例)进行染色体核型分析。结果56例CML患者有5例为细胞培养失败,均为慢性期患者,该5例细胞培养失败病例经FISH技术证实均出现Ph 染色体,余46例慢性期患者中有42例出现Ph 染色体,其中2例为双Ph 染色体;3例合并有复杂的染色体变化,包括染色体数目及结构的异常,分别为-2,-6,-19,-16,-20,-Y, 8以及t(2;4)(p16;p15)。CML慢性期共有47例患者出现Ph 染色体,总阳性率为92.17%。5例加速及急变期患者均出现Ph 染色体,其中3例患者合并有复杂的染色体核型变化:-Y,del(10),I(9)。结论染色体核型分析对CML的疾病分期、进展、治疗及预后有重要的临床意义。FISH技术在CML染色体核型分析上可作为重要技术补充手段弥补CC检查的不足。     

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     

Heterogeneous prognostic impact of derivative chromosome 9 deletions in chronic myelogenous leukemia

Derivative chromosome 9 deletions are seen in 10% to 15% of patients with chronic myelogenous leukemia and have been associated with a poor prognosis; however, no studies have been performed in the context of a randomized clinical trial. We developed a DNA-based deletion screen and investigated 339 chronic phase patients treated with interferon-alpha as first-line therapy in 3 controlled German studies with a median observation time of 7 years. Deletions were detected in pretreatment DNA of 59 of 339 (17%) patients. Of these, 21 spanned the ABL/BCR junction and 38 were centromeric (n = 20) or telomeric (n = 18) of the breakpoint. There was no significant difference in overall survival between deleted and nondeleted patients. Patients with breakpoint-spanning deletions had poorer survival compared with patients without deletions (4.7 versus 7.8 years; P = .003), but this was not significant when censored at allogeneic stem cell transplantation (n = 129) or imatinib (n = 62) treatment in the first chronic phase (P = .078). Unexpectedly, deletions that did not span the breakpoint were associated with improved survival compared with cases without deletions (P = .001). Multiple Cox regression analysis indicated that deletion status (P = .007), age (P = .018), and spleen enlargement (P < .001) were significant independent indicators of survival and confirmed that only deletions spanning the ABL/BCR breakpoint were associated with an adverse prognosis (P = .039).     

Imatinib mesylate therapy may overcome the poor prognostic significance of deletions of derivative chromosome 9 in patients with chronic myelogenous leukemia

Deletions of derivative chromosome 9 [der(9)] can be identified by fluorescence in situ hybridization (FISH) in 10% to 15% of patients with chronic myeloid leukemia (CML). Patients with der(9) deletions have been reported to have an adverse outcome when treated with chemotherapy, interferon, and possibly imatinib mesylate. We investigated the frequency and prognostic significance of der(9) deletions among 352 patients with CML treated with imatinib mesylate at our institution, in whom a deletion status of der(9) was determined. Thirty-three patients (9%; 95% CI 0.07, 0.13) (30 in chronic phase, 3 in accelerated phase) had der(9) deletions. The rates of major (82% vs 79%, P = 0.82) and complete cytogenetic response (76% vs 66%, P = .33) with imatinib mesylate therapy were similar in patients with and without der(9) deletions, respectively. After a median follow-up of 28 months, there was no difference in overall survival (P = .30) or response duration (P = .49) in patients with and without deletions. In a multivariate analysis, der(9) deletions had no significant impact on response, survival, or response duration. We conclude that treatment with imatinib mesylate overcomes the adverse prognostic significance of der(9) deletions in patients with CML.     

Double jeopardy from a single translocation: deletions of the derivative chromosome 9 in chronic myeloid leukemia

Chronic myeloid leukemia (CML) is characterized by formation of a BCR-ABL fusion gene, usually as a consequence of the Philadelphia (Ph) translocation between chromosomes 9 and 22. Recently the development of new fluorescence in-situ hybridization (FISH) techniques has allowed identification of unexpected deletions of the reciprocal translocation product, the derivative chromosome 9, in 10% to 15% of patients with CML. These deletions are large, span the translocation breakpoint, and occur at the same time as the Ph translocation. Such deletions therefore give rise to previously unsuspected molecular heterogeneity from the very beginning of this disease, and there is mounting evidence for similar deletions associated with other translocations. Several studies have demonstrated that CML patients who carry derivative chromosome 9 deletions exhibit a more rapid progression to blast crisis and a shorter survival. Deletion status is independent of, and more powerful than, the Sokal and Hasford/European prognostic scoring systems. The poor prognosis associated with deletions is seen in patients treated with hydroxyurea or interferon, and preliminary evidence suggests that patients with deletions may also have a worse outcome than nondeleted patients following stem cell transplantation or treatment with imatinib. Poor outcome cannot be attributed to loss of the reciprocal ABL-BCR fusion gene expression alone, and is likely to reflect loss of one or more critical genes within the deleted region. The molecular heterogeneity associated with the Philadelphia translocation provides a new paradigm with potential relevance to all malignancies associated with reciprocal chromosomal translocations and/or fusion gene formation.     

Size matters: the prognostic implications of large and small deletions of the derivative 9 chromosome in chronic myeloid leukemia

Deletions of the derivative 9 chromosome (der(9)) are associated with poor prognosis in chronic myeloid leukemia (CML). Several models have been proposed to account for this association. To distinguish between the various models we mapped the deletion in 69 Philadelphia-positive CML patients carrying a der(9) deletion and compared the size of the deletion with the patients' outcome. Our results demonstrate that patients with large deletions had a significantly worse survival than those with small deletions whereas the outcome for patients with small deletions was similar to that of patients lacking a deletion. These results support the tumor suppressor gene model for the pathogenesis of der(9) deletions, argue against alternative models and provide insight into candidate gene location.