急性粒-单核细胞白血病的细胞遗传学异常研究

背景与目的：2001年WHO分型特别提出4种伴再现性遗传学异常的急性髓细胞白血病（AML），其中inv（16）（p13：q22）与急性粒一单核细胞白血病（M4）密切相关，是预后好的标志。本研究旨在探讨M4的细胞遗传学特征。方法：采用直接法及短期培养法制备骨髓细胞染色体，并以R显带技术对89例M4患者进行核型分析，并应用间期荧光原位杂交（I-FISH）技术对其中伴有＋22异常的患者进行inv（16）检测。结果．89例M4患者中，异常染色体检出率为40．4％（36／89），共12种主要异常核型，其中5种为特异性染色体异常，见于25例患者，占核型异常患者的69．4％。单纯＋8（10例）为最常见的数目异常；结构异常最多见的是inv（16）（5例）；t（8；21）者3例；伴t（9；22）者有1例；其中5例inv（16）及3例＋22均只见于M4Eo患者。3例＋22患者FISH检测inv（16）均阳性。结论：细胞遗传学研究对于急性粒一单核细胞白血病的诊断具有重要价值，但是细胞遗传学对inv（16）检测阳性低，对怀疑病例及所有＋22异常的患者，应尽可能进行FISH检测。     

急性粒-单核细胞白血病的免疫表型及细胞遗传学特征

背景与目的:白血病中某些抗原的表达与细胞遗传学的改变密切相关。本研究旨在探讨急性粒-单核细胞白血病(acutemyelomonocyticleukemia,M4)的免疫表型特征,并分析其与细胞遗传学的关系,为其诊断及治疗提供依据。方法:采用一组系列相关单克隆抗体和三色流式细胞术对81例M4患者进行免疫表型分析,并分析有核型资料的73例患者的核型异常;同时应用双色间期荧光原位杂交(fluorescenceinsituhybridization,FISH)技术对有染色体标本的35例患者进行inv(16)检测,并分析inv(16)阳性患者的免疫表型特征。结果:81例M4患者中48例(59.3%)表达干细胞标志CD34;髓系标志以CD33(84.0%)表达最高,其次为CD13(81.5%)和CD14(24.7%);分别有23例(28.4%)和10例(12.3%)患者表达T系和B系相关抗原。11例M4Eo患者中CD13均阳性,10例CD33阳性,7例CD34阳性,5例CD2阳性。FISH检测发现14例患者inv(16)阳性,其中CD13阳性13例,CD33阳性11例,CD34阳性8例,CD14阳性5例,CD7阳性3例,CD2阳性3例。73例M4患者中,异常染色体检出率为41.1%(30/73),共检出11种主要异常核型。CD2和CD34在染色体异常患者中表达明显增高,而CD14表达减低。CD2在M4Eo中表达增高,但其表达与inv(16)无显著性相关。结论:急性粒-单核细胞白血病以髓系抗原表达为主,CD2和CD34在染色体异常患者中表达明显增高;CD2在M4Eo中表达增高。Inv(16)在M4中占40%,各抗原表达与inv(16)无显著相关性。     

Ph-bcr-abl+血液病的间期荧光原位杂交检测

 目的 探讨间期荧光原位杂交（FISH）技术在检测Ph- bcr-abl+血液病患者中的作用。方法 应用bcr-abl双色探针双融合荧光原位杂交（D-FISH）技术对7例常规细胞遗传学（CC）检测Ph-或不肯定而巢式反转录-聚合酶链反应（RT-PCR）检测bcr-abl+的血液病患者进行DNA水平的bcr-abl融合基因检测。结果 例2 FISH检测第一次bcr－abl+细胞占13.5 %,第二次bcr-abl+细胞占11.5 %。例4和例6分别检出1 %的bcr-abl+细胞,略高于正常分界值。例7检出0.5 %的bcr-abl+细胞,其余患者未检出bcr-abl+细胞。结论 Ph- bcr-abl+血液病患者DNA水平存在低水平的bcr-abl+细胞;FISH有助于Ph-或不确定以及bcr或abl断裂点不在通常位点的bcr-abl+细胞的检出。     

联合应用细胞遗传学、巢式RT-PCR和FISH技术监测慢性髓细胞白血病干扰素治疗中肿瘤负荷

 目的　探讨常规细胞遗传学（CC）、巢式反转录-聚合酶链反应（RT-PCR）及双色双融合荧光原位杂交（D-FISH）三种技术监测慢性髓细胞白血病（CML）患者干扰素（INF）治疗过程中肿瘤负荷的灵敏度和特异性。方法　联合应用CC、巢式RT-PCR 和D-FISH三种技术对5例INF治疗中CML患者的肿瘤负荷水平进行检测。结果　对5例CML患者INF治疗前后的24份骨髓标本检测显示,23份标本检出不同比率的Ph染色体,所有标本RT-PCR结果均为阳性。病例2治疗后75个月Ph- bcr-abl+标本经D-FISH检测,仍有4.5 %的bcr-abl+细胞;病例1治疗后22、26个月的2份标本,病例5治疗后12、16个月的2份标本及病例4治疗后6、10个月的2份标本经CC检测Ph染色体阳性率相同,进一步行D-FISH检测,分别检出47.5 %和39.5 %、74.0 %和60.5 %、99.0 %和99.5 %的bcr-abl+细胞。结论　CC可作为监测CML患者治疗过程中肿瘤负荷水平的基本手段。在CC检测结果无法准确判断体内肿瘤负荷动态变化时以及治疗患者体内肿瘤负荷降低到CC不能检出而RT-PCR仍为阳性时,需借助FISH来准确检测体内肿瘤负荷,监测其动态变化。FISH检测bcr-abl转阴的患者需靠灵敏度更高的RT-PCR监测。     

三体22在预测inv(16)急性髓细胞白血病中的重要意义

目的：探讨三体22在诊断inv(16)急性髓细胞白血病(AML)中的意义。方法：分别采用Spectrum Green标记的22号染色体着丝粒DNA探针(CEP22)1和以红、绿荧光素直接标记的MYHll基因断裂点分开的两侧序列作为探针。对1例伴inv(16)／ 22克隆异常的AML及1例单纯 22克隆异常的AML进行单色间期FISH和D—FISH检测．并与细胞形态学及常规细胞遗传学(CC)相比较。结果：2例经采用CEP22探针的FISH技术证明确实存在三体22克隆性异常，而D-FISH均证实inv(16)阳性。其中1例的少数间期核(1．6％)同时伴有16p13的微小缺失。结论：由于 22是一种染色体数目异常，相对来说易于辨认，可以看做是预报inv(16)AML的重要信号，具有潜在的诊断inv(16)AML的价值。     

急性髓系白血病16号染色体倒位的研究进展

 inv(16)是急性髓系白血病（AML）M4Eo的特征性的异常核型,是预后较好的一个标志。inv(16)的结果是16q22上的CBFb基因和 16p13上的MYH11基因重排,产生CBFb-MYH11融合基因,可能在白血病的发病机制中起重要作用。本文就inv(16) AML的临床、血液学特点及分子生物学特征等方面的研究进展作一综述。     

霍奇金淋巴瘤治疗后继发急性髓系白血病伴染色体核型inv(16)一例并文献复习

目的 提高对治疗相关白血病的认识.方法 报道1例霍奇金淋巴瘤(HL)治疗后继发急性髓系白血病(AML)伴inv(16)患者的诊治过程,并结合文献复习治疗相关白血病的发病机制、治疗及预后.结果 该例36岁女性患者在确诊HL并接受规范淋巴瘤化疗后2年确诊为治疗相关白血病.染色体核型分析显示inv(16),且聚合酶链反应(PCR)结果提示CBFβ/MYH11融合基因阳性.经3个疗程化疗后,患者CBFβ/MYH11基因仍为阳性.该患者行半相合外周血造血干细胞移植后取得分子生物学缓解.患者一般情况良好,现无白血病生存时间1.5年.结论 治疗相关白血病治疗难度大,但其具有异质性.细胞遗传学及分子生物学对于治疗相关白血病预后有重要的提示意义.目前异基因造血干细胞移植为唯一可以治愈治疗相关性AML的有效方式.     

CD_4抗原在急性髓系白血病细胞上的表达及其意义

目的 :探讨 CD4抗原在急性髓系白血病细胞上的表达及意义。方法 :对 82例急性髓系白血病患者进行免疫表型、细胞遗传学分析。结果 :CD4在 AML患者中表达率为 40 .2 % ,M5 中阳性率最高 92 .9% ,M4次之为5 5 % ,CD+4 AML 高表达 HL A- DR,CD3 8、CD3 3 、CD1 5 、CD1 4 、TB系列抗原阴性。CD+4 AML 中可见 11q2 3、inv(16 )、t(9;2 2 ) ,未见特异性染色体异常。但伴 11q2 3和 inv(16 )异常的 AML频繁表达 CD4,阳性率分别为 86 .3%、6 0 .2 %。CD+4和 CD- 4 AML 在年龄、性别、肝脾肿大、CNS- L、DIC、1疗程 CR率等临床特征方面无明显不同。结论 :CD+4 AML 是一组起源较早的具有单核细胞特征的髓系白血病 ,CD4的表达对 AML- M4、M5 尤其是 M5 亚型的鉴别诊断有重要价值     

t(8;21)急性髓系白血病患者MICM分型及预后的回顾性分析

 目的　分析t(8;21)急性髓系白血病（AML）患者的细胞形态学、免疫表型、遗传学、分子生物学（MICM）分型及临床治疗疗效。方法　运用瑞特染色法、FAB细胞形态分类标准、流式细胞术（FCM）直接免疫荧光标记技术、遗传学染色体吉姆萨显带技术及RT-PCR技术对70例确认有t(8;21)与AML1-ETO融合基因双阳性的AML患者及70例正常染色体核型的AML患者进行分析和比较。结果　70例t(8;21)AML患者中M1 1例,M2 64例,M4 3例,无法分型的急性白血病（AL）2例;免疫表型分析发现CD13、CD33、CD34、CD117高表达,40 %表达CD19,11 %表达CD15,10 %表达CD11b,7 %表达CD7;遗传学显示50 %的t(8;21) AML患者有附加染色体异常,主要为性染色体丢失、9q-及超二倍体;RT-PCR检测AML1-ETO融合基因100 %阳性。CD+19 t(8;21) AML患者完全缓解（CR）率72 %,CD+19伴CD+7 t(8;21)AML患者CR率为0,正常核型CR率31 %。结论　t(8;21) AML患者主要在M2中集中出现,附加染色体异常较多见。CD19表达较高,而CD7表达极低,CD34、CD117高表达,这些抗原的表达可能与核型密切相关。CD+19 是预后良好的指标,但同时出现CD+7,则预后不良。     

伴3'RARα亚显微缺失的急性早幼粒细胞白血病一例

 目的　报道1例伴RARα 3'-末端（3'RARα）亚显微缺失的M3r亚型急性早幼粒细胞白血病（APL）病例及其形态学、细胞遗传学、分子遗传学和分子生物学的研究结果。方法　骨髓细胞直接法和短期培养法制备染色体,应用反带技术进行核型分析;分别用CEP X／Y α-卫星DNA探针、LSI PML-RARα 双色双融合探针和LSI RARα 双色断裂点分离探针进行荧光原位杂交（FISH）分析;实时定量反转录聚合酶链反应（RT-PCR）方法检测PML-RARα 融合基因转录本;多重巢式RT-PCR技术检测急性白血病29种染色体畸变所形成的融合基因,包括PML-RARα,PLZF-RARα和NPM-RARα融合基因转录本。结果　反带分析显示核型为45,X,-Y[6]／46,XY[8],CEP X／Y探针FISH进一步证实了Y染色体丢失;RARα双色断裂点分离探针FISH分析显示1个RARα等位基因的整个3'-末端缺失;通过细胞遗传学、FISH以及RT-PCR等方法检测,排除PML-RARα、PLZF-RARα、NPM-RARα、NuMA-RARα和STAT5b-RARα重排。结论　识别APL中一种新的RARα 基因重排类型（3'RARα 亚显微缺失而无X-RARα 融合）,RARα 双色断裂点分离探针FISH分析是明确该异常的有效手段,其分子学结果有待进一步阐明。     

Abnormalities of chromosome 16q in myeloid malignancy: 14 new cases and a review of the literature.

Fourteen patients with abnormalities of chromosome 16q, 13 with acute myelogenous leukaemia (AML), and one with refractory anaemia with excess of blasts (RAEB), are described. Seven patients had inv(16)(p13q22), two had del(16)(q22), and five had other abnormalities of 16q. Six of the seven patients with inv(16) had AML M4Eo and, following treatment with adriamycin, cytosine arabinoside, and 6-thioguanine, all achieved complete remission (CR). Neither patient with del(16)(q22) had typical M4Eo morphology at diagnosis; CR was achieved in one and one had resistant leukaemia. Patients with other abnormalities of 16q had blasts of diverse morphology and, although morphologically abnormal eosinophils were seen in three patients, this was not as marked as in the patients with inv(16). CR was achieved in two of the four patients with other abnormalities of 16q but duration of remission was short in both cases. These results suggest that most patients with del(16)(q22) and other abnormalities of 16q22 do not have typical AML M4Eo. Such patients tend to have a worse prognosis, and are more likely to have complex karyotypes typical of secondary leukaemia.     

间期荧光原位杂交检测血液病染色体三体8

目的 探讨间期荧光原位杂交(FISH)技术在检测血液病染色体三体8中的价值.方法 用荧光素直接标记的8号染色体着丝粒探针对77例血液病患者进行间期FISH检测,并与常规细胞遗传学方法(CC)进行比较分析.结果 FISH法三体8的检出率较传统吉姆萨显带高,或在吉姆萨显带无法分析时提供结果.结论 FISH检测三体8的敏感性高于常规核型分析,在小克隆检测方面有其优越性.     

Clonal chromosomal abnormalities in the stem cell compartment of patients with acute myeloid leukemia in morphological complete remission.

Acute myeloid leukemia arises from the clonal expansion of a malignant transformed progenitor cell. Despite intensive chemotherapy, final disease eradication is achieved by a small proportion of cases only and 50-70% of adults with AML will ultimately relapse and die from their disease. Hence residual disease below the level of morphological detectability must be assumed in clinical and morphological complete remission. CD34+/CD38- and CD34+/CD38+ subpopulations of seven patients in morphological complete remission were isolated by FACS (purity >98%) and were analyzed by conventional cytogenetics or FISH for chromosomal aberrations. In five of seven patients, clonal chromosomal abnormalities were detected in the CD34+/CD38+ subpopulation and in one patient with AML M2 (add (2)(q37)) in the most immature CD34+/CD38- stem cell compartment. One patient with AML M4Eo (inv(16),+8), showed a normal karyotype by conventional cytogenetic analysis, whereas four of 15 metaphases of the sorted CD34+/CD38+ subpopulation revealed the inversion 16. These observations underline that leukemic cells can survive intensive chemotherapy in the niche of the stem cell compartment. In some patients the sensitivity for the detection of persistent leukemic cells seems to be higher in FACS-sorted subpopulations than conventional cytogenetic analysis of the unseparated bone marrow. Immunophenotyping revealed minimal residual disease in four of the patients. Functional analysis has to be performed to investigate the leukemogenic potential of these residual cells.     

Chromosome change at 16q22 in nonlymphocytic leukemia: clinical implication on leukemia patients with inv(16) versus del(16)

Nineteen patients with inv(16)(p13q22) or del(16) in myeloid leukemia are described. Eight showed inv(16)(p13q22), including one with de novo acute myeloid leukemia (AML-M2) and seven with de novo acute myelomonocytic leukemia (AMML-M4). Additional chromosome changes were detected in five of the cases; the most common change was trisomy 22. All but one of the de novo M2 and M4 leukemia patients with inv(16)(p13q22) showed initial bone marrow eosinophilia (greater than 5%) with basophilic granules. The remaining 11 showed deletion of the long arm of a chromosome no. 16 [del(16)(q22 or q23)]. Eight of the 11 were diagnosed as having chronic myelomonocytic leukemia, three transformed into an acute phase with M4 morphology; none of them gained complete remission. Two of the remaining three patients with del(16) were diagnosed as having M4 leukemia without marrow eosinophilia. The remaining one was a case of M4 leukemia following a myelodysplastic syndrome. The findings indicate that del(16) might be related to chronic myelomonocytic leukemia or leukemia with a prior history of myelodysplastic syndrome without evidence of marrow eosinophilia. On the other hand, inv(16)(p13q22) is highly associated with de novo AML especially AMML-M4 with bone marrow eosinophilia and a favorable prognosis.     

Individual patient data-based meta-analysis of patients aged 16 to 60 years with core binding factor acute myeloid leukemia: a survey of the German Acute Myeloid Leukemia Intergroup.

PURPOSE: To evaluate prognostic factors for relapse-free survival (RFS) and overall survival (OS) and to assess the impact of different postremission therapies in adult patients with core binding factor (CBF) acute myeloid leukemias (AML). PATIENTS AND METHODS: Individual patient data-based meta-analysis was performed on 392 adults (median age, 42 years; range, 16 to 60 years) with CBF AML (t(8;21), n = 191; inv(16), n = 201) treated between 1993 and 2002 in prospective German AML treatment trials. RESULTS: RFS was 60% and 58% and OS was 65% and 74% in the t(8;21) and inv(16) groups after 3 years, respectively. For postremission therapy, intention-to-treat analysis revealed no difference between intensive chemotherapy and autologous transplantation in the t(8;21) group and between chemotherapy, autologous, and allogeneic transplantation in the inv(16) group. In the t(8;21) group, significant prognostic variables for longer RFS and OS were lower WBC and higher platelet counts; loss of the Y chromosome in male patients was prognostic for shorter OS. In the inv(16) group, trisomy 22 was a significant prognostic variable for longer RFS. For patients who experienced relapse, second complete remission rate was significantly lower in patients with t(8;21), resulting in a significantly inferior survival duration after relapse compared with patients with inv(16). CONCLUSION: We provide novel prognostic factors for CBF AML and show that patients with t(8;21) who experience relapse have an inferior survival duration.