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。     

一例伴t(14;14)(q11;q32)易位的B细胞急性淋巴细胞白血病的临床和分子细胞遗传学研究

目的 报告1例伴t(14;14)(q11;q32)易位的罕见B细胞急性淋巴细胞白血病(B-lineage acute lymphoblastie leukemia,B-ALL)病例,阐明其临床和分子细胞遗传学特征.方法 分析1例伴t(14;14)(q11;q32)易位B-ALL患者的临床资料;将患者骨髓细胞24h培养后按常规方法制备染色体标本,采用R显带技术进行核型分析;分别应用IGH双色断裂点分离探针、CEBPE双色断裂点分离探针、4号全染色体涂染探针和ALL组合探针进行荧光原位杂交(fluorescence in situ hybridization,FISH)分析.结果 常规细胞遗传学分析显示患者核型为47,XX,+4,t(14;14)(q11;q32)[20],FISH分析进一步证实了这种核型异常.IGH双色断裂点分离探针FISH分析表明t(14;14)(q11;q32)易位累及IGH基因,CEBPE双色断裂点分离探针FISH分析提示t(14;14)(q11;q32)易位中IGH的伙伴基因为CEBPE基因.结论 在B-ALL中t(14;4)(q11;q32)易位同时累及IGH和CEBPE基因为少见的再现性遗传学异常,该异常可定义B-ALL中一种新的亚型.伴有t(14;14)(q11;q32) IGH/CEBPE易位的B-ALL患者可能预后较好.     

慢粒患者经格列卫治疗后细胞遗传学改变:标准Ph易位t(9;22)(q34;q11)变为变异Ph易位t(21;22)(p11;q11)

目的通过对慢性粒细胞白血病(chronic myeloid leukemia,CML)患者经格列卫药物治疗后细胞遗传学改变的研究,探讨ABL-BCR的表达缺失与获得性格列卫耐药的关系。方法应用R显带技术对染色体进行核型分析,并选用BCR/ABL探针,通过双色荧光原位杂交技术进一步确认遗传学分析。结果患者经格列卫治疗后染色体核型由t(9;22)(q34;q11)变为t(21;22)(p11;q11)。双色荧光原位杂交证实此患者核型应为46,XY,t(9;22;21)(q34;q11;p11)。结论变异Ph易位中ABL-BCR的表达缺失与获得性格列卫耐药有关,荧光原位杂交技术在检测变异易位中起重要作用。     

5例伴有t(16;21)(p11;q22)急性白血病的临床和实验研究

目的：报告5例伴有t(16;21)(p11;q22)的急性白血病和其中1例的染色体涂染分析。方法：骨髓细胞24h培养后按常规方法制备染色体，采用R显带技术进行染色体核型分析，并以16号和21号整条染色体涂染探针对其中1例患者进行染色体涂染检测。结果：5例均显示t(16;21)(p11;q22),占15年来进行染色体检查的急性非淋巴细胞白血病患者总数的0．3％（5／1448）。5例均无白血病细胞吞噬其他血细胞现象。1例患者的染色体涂染分析证实了16号和21号染色体之间发生了相互易位。结论：t(16;21)是急性非淋巴细胞白血病中1种少见的非随机的染色体易位，代表了1种独特的白血病亚型。染色体涂染技术是比常规核型分析更为可靠的检测该易位的手段。     

伴有22三体的t(5;17)变异易位急性早幼粒细胞白血病

目的 探讨一例核型为47,XY,t(5;17),+22的少见急性早幼粒细胞白血病(acute promyelo-cytic leukemia,APL)的临床和实验特征.方法 在常规核型分析的基础上,应用荧光原位杂交(fluorescencein situ hybridization,FISH)和多重荧光原位杂交(multiplex fluorescence in situ hybridization,M-FISH)技术进一步检测该病例的细胞遗传学异常,并结合文献分析此类少见变异易位的临床特点.结果 FISH检测PML-RARa阴性,但77%的细胞显示存在17号RARa基因的重排或复制;BCR-ABL阴性,但74%的细胞显示有22号染色体的复制或重排.M-FISH明确RARa基因重排系5号与17号染色体易位所致,并证实了22号三体的存在.结论 变异型t(5;17)易位,形成NPM-RARa融合基因的急性早幼粒细胞白血病是APL中少见的类型.骨髓形态表现为奥氏小体缺如,核型中常伴有其它附加染色体异常,全反式维甲酸(all-trans retinoicacid,ATRA)联合化疗有效,但易复发,合并弥漫性血管内凝血及高白细胞者预后凶险.     

荧光原位杂交技术及染色体核型分析在罗伯逊易位型21-三体检测中的应用

目的 通过对孕妇羊水进行染色体核型、荧光原位杂交(fluorescence in situ hybridization,FISH)分析,探讨两者联合检测在诊断罗伯逊易位型21-三体中的应用价值.方法 为2010年1月至2011年12月进行产前诊断的孕妇抽取羊水,经体外细胞培养后进行G显带染色体核型分析.对发现的5例罗伯逊易位采用FISH检测间期细胞13、18、21及X/Y的染色体数目,并分析孕妇及其丈夫外周血染色体核型.结果 两个胎儿父母外周血染色体核型正常,其中一个胎儿羊水染色体核型为46,XY,rob(21;21)(q10;q10),FISH检测提示其为21-二体,另一个胎儿核型为46,XY,rob(14;21)(q10;q10),FISH检测证实其为21-三体.另外3个胎儿母亲外周血染色体核型分别为45,XX,rob(14;21)(q10;q10)、45,XX,rob(15;21)(q10;q10)、45,XX,rob(21; 22) (q10; q10),其羊水染色体核型分别为46,XX,rob(14;21)(q10;q10)、46,XY,rob(15;21) (q10;q10)、46,XX,rob(21;22)(q10;q10).FISH检测证实其均为21-三体.结论 染色体核型分析结合FISH检测有助于明确罗伯逊易位型21-三体的诊断,但FISH检测同源罗伯逊易位型21-三体征有一定局限性.     

47,XY,t(2;21) (q21;q22.3),+21一例

患儿 男,5个月.因感冒检查发现心脏杂音入院.入院心脏超声检查示先天性心脏病:动脉导管未闭、卵圆孔未闭、右室肥大、轻度肺高压.同时发现患儿眼裂小、眼角外侧上翘、眼间距宽等特殊面容.细胞遗传学检查:取外周血常规淋巴细胞培养、制片、G显带.用染色体分析系统AI染色体核型分析工作站的Cytovision分析系统,计数50个中期分裂相,完整分析20个分裂相,发现该患儿的染色体为罕见易位型21三体,核型为:47,XY,t(2;21)(2pter→2q21::21q22.3→21qter;21pter→21q22.3::2q21→2qter),+21(图1).其父母核型正常.     

染色体复杂重排的细胞遗传学检测及遗传咨询

目的 以4例染色体复杂重排新核型的确诊为例,探讨这类染色体异常的检测方法及遗传咨询。方法 应用常规G显带技术分析4例复杂易位患者的染色体核型,其中2例为智力低下患者,另2例来自有自然流产史的夫妇。2例智力低下患者应用FISH和CGH技术进一步分析并检测其父母核型。查询相关数据库检索4例核型的发生率。结果 4例患者的核型分别为46,XYqh+,t(1;12;2;10)(q25;q11;p14;p11),inv(1)(p22q25),46,XY,t(7;21;8)(p13;q22;p21),46,XX,t(3;7;10)(q28;p15;q22)和46,XY,t(2;16;5)(q33;p12;q33)。2例有智力低下患者经FISH和CGH检测未发现其他染色体的异常,未见染色体微小重复或缺失。4例核型均为国内外文献未曾报道的新核型。结论 染色体复杂重排的遗传学检测需要综合考虑多种核型分析方法的结果,染色体复杂重排的遗传咨询需要着重结合其重排类型和临床症状进行分析。     

急性淋巴细胞白血病的t(1;19)(q21;q13)易位

在Kaneko等(1982年)报道的许多急性淋巴细胞白血病(ALL)的患者中,一例非B,非T ALL的3岁女孩,在诊断和复发时都发现有相互易位的改变,t(1;19)(q21;q13)。在第三届白血病染色体专业会议的330例ALL患者中,她是唯一具有此种易位的病例。作者发现一例患有非B,非T高危ALL的22个月的男孩,也具有同样易位。诊断时,其核型为46,XY/47,XY,-F, 2C;复发时,核型为47,XY, 8,-19, t(1;(19)(q21;q13);inv(5)(q13q35)。该病例的t(1;19)易位,系第一号染色体部分三体所致。     

Characterization of the human myeloid leukemia-derived cell line GF-D8 by multiplex fluorescence in situ hybridization, subtelomeric probes, and comparative genomic hybridization.

The human myeloid leukemia cell line GF-D8 was established from the peripheral blood blasts of a patient with acute myeloid leukemia FAB subtype MI (AML-MI). The karyotype, which has not changed significantly over several years of culture, was described initially as 44,XY,-5,del(7q),inv(7q),add(8q),add(11q),del(12p),-15,-17,+mar. With the advent of multicolor fluorescence in situ hybridization (FISH) techniques, the prospect of accurately characterizing this complex karyotype became feasible. In the present study, we applied 24-color whole-chromosome painting and analyzed the results using a filter-based detection system and proprietary software for multiplex FISH (M-FISH). This resulted in the refinement of the karyotype and the identification of hitherto unsuspected chromosome rearrangements. M-FISH identified the origin of the add(8q) and add(11q) as well as the small marker chromosome. Both the del(7q) and del(12p) were redefined as unbalanced translocations and an apparently normal chromosome 11 was shown to be t(11;17). Importantly, the del(12p) was shown to be a der(12)t(7;12). Single-color whole-chromosome painting studies confirmed these findings, but also identified a cryptic t(Y;12) not seen in the original M-FISH analysis. We then carried out a FISH screening assay using a complete set of chromosome-specific subtelomeric probes. This allowed the identification of p and q subtelomeric regions involved in the translocations and indicated amplification of the 8q subtelomeric region. Comparative genomic hybridization (CGH) revealed a highly unbalanced karyotype, as deletions accompanied the majority of translocations, and identified the regions of amplification as 8q22.3-qter and 11q21-qter. Finally, conventional FISH with centromeric and unique sequence probes was necessary to elucidate all of the rearrangements.     

Molecular characterization of der(15)t(11;15) as a secondary cytogenetic abnormality in acute promyelocytic leukemia with cryptic PML-RAR alpha fusion on 17q

A case of acute promyelocytic leukemia (APL) with cryptic PML-RAR alpha fusion on 17q and add(15p) as a secondary abnormality was characterized using molecular cytogenetic techniques. Spectral karyotyping (SKY) showed that chromosome 11 material was added to 15p, forming a der(15)t(11;15), which was refined to der(15)t(11;15)(q13.2;p13) with information obtained by comparative genomic hybridization (CGH). Interstitial insertion of chromosome 15 material into chromosome 17q was found by fluorescence in situ hybridization (FISH) with whole chromosome painting (WCP) probes. This study illustrates the necessity of a combination of molecular cytogenetics to decipher complex karyotypic abnormalities and cryptic translocations in leukemia.     

Unbalanced cryptic 5p deletion/17p duplication identified by subtelomeric FISH in a family with a boy with chimerism and a balanced t(4;5)

The use of subtelomeric FISH probes has greatly supplemented conventional chromosome analysis in detecting cryptic anomalies in patients with mental retardation (MR), dysmorphic features, and congenital malformations. We report a 3-month-old boy who was diagnosed with ambiguous genitalia, dysmorphic features, and developmental delay. Standard chromosome studies on blood revealed a chimeric karyotype of 46,XY,t(4;5)(q31.1;q14)[46]/46,XX[4]. The boy had intra-abdominal gonads that were testicular in origin by biopsy. Multiple dysmorphic features, marked hypotonia, developmental delay, poor growth, and relative macrocephaly were noted on physical exam. His 2.5-year-old sister also presented with hypotonia, developmental delay, relative macrocephaly, and similar dysmorphic stigmata. In addition, she was diagnosed with several internal malformations. Her karyotype was 46,XX. Due to the striking phenotypic similarity, subtelomeric FISH studies were initiated in the siblings. In addition to the known balanced karyotypic abnormalities, the boy was found to have a derivative chromosome 5 with a 5pter deletion and a 17pter duplication. This cryptic abnormality was also detected in his sister. Chromosome analysis of the father revealed a subtle balanced t(5;17)(p15.31;p13.1) which was confirmed by subtelomeric FISH, whereas the mother's chromosome complement was normal. This familial constellation illustrates the usefulness of subtelomeric FISH in the diagnosis of cryptic chromosome abnormalities in patients for whom conventional karyotype does not disclose findings sufficient to explain the observed phenotypic anomalies.     

一例伴有der(Y)t(Y;1)异常的多发性骨髓瘤的临床和实验研究

目的 对1例伴有不平衡染色体易位der(Y)t(Y;1)的多发性骨髓瘤(multiple myeloma,MM)患者进行细胞遗传学、中期荧光原位杂交、免疫学及临床研究.方法 采用细胞遗传学G显带行中期染色体核型分析;用1号染色体涂抹探针、Y染色体异染色质区探针进行中期荧光原位杂交检测;免疫分型检测CD38、CD138、ZAP70等的表达及免疫电泳检测免疫球蛋白类型等.结果 细胞遗传学分析结果 发现患者具有高度复杂的异常克隆,其核型为:92,XXYY[3]/49,X,der(Y)t(Y;1)(q12;q21),t(11;14)(q13;q32),+18,+20,+21[47]/49,X,idem,del(13q22),ace[1]/98,XX,der(Y)t(Y;1)×2,+18,+18,+20,+20,+21,+21[10]/46,XY[19].中期荧光原位杂交结果 证实der(Y)t(Y;1)的G显带结果 ,为1q部分三体与Y染色体长臂的不平衡易位.其异常的单克隆免疫球蛋白为IgD,定量6.24 g/L;免疫分型结果 为表达CD38、CD138,不表达ZAP70,考虑为异常克隆浆细胞的表达.结论 Y染色体的结构异常在血液系统肿瘤中非常罕见,本文报道1例发生于多发性骨髓瘤中的伴der(Y)t(Y;1)的核型异常、实验室及临床特点.     

Interstitial deletion of chromosome 4, del(4)(q12q21.1), in a mentally retarded boy with a piebald trait,due to maternal insertion,ins(8;4)

A 17-year-old boy who was diagnosed with “Waardenburg syndrome” showed moderate growth and mental retardation. Chromosome analysis showed an apparent interstitial deletion 4q12q21.1. The mother had a direct insertion of the deleted segment into a chromosome 8. The rearrangement was confirmed to be nonreciprocal and an insertion by in situ hybridization using whole chromosome 4 and 8 painting probes. The mother's karyotype is 46,XX,ins(8;4)(q21.2; q12q21.1); that of the propositus is 46,XY, der(4)ins(8;4)(q21.2;q12q21.1)mat. This is the first report of an inherited proximal 4q deletion. Am. J. Med. Genet. 75:78–81, 1998. © 1998 Wiley-Liss, Inc.     

A new variant translocation 11;17 in a patient with acute promyelocytic leukemia together with t(7;12)

Bone marrow cells from the majority of patients with acute promyelocytic leukemia (APL) are characterized by t(15;17)(q22;q11-12). At least 12 variant translocations have both also reported, and in each case, either abnormal chromosome 15 or del(17q) or both were involved in complex rearrangements. We report a patient with APL showing two translocations without apparent involvement of chromosome 15 and without del(17q). The karyotype was 46,XY,t(7;12)(p15;p13),t(11;17)(q13;q12). Rearrangement involving t(11;17) is probably associated with APL, while t(7;12) appears to be therapy related.