Cytogenetic characterization of diffuse large cell lymphoma using multi-color fluorescence in situ hybridization

We have employed multi-color fluorescence in situ hybridization (M-FISH) to characterize the cytogenetic changes in 20 diffuse large B-cell lymphomas (DLBCL), that contained complex and partially characterized karyotypes. The M-FISH analysis helped to delineate 94% of the unidentified abnormalities and assisted in redefining some unidentified/misidentified karyotypic changes. Recurrent breakpoints observed in approximately 20% cases included 14q32, 3p21, 3q27, 22q12, 1q25, and 18q21 (in decreasing order), and 1p22, 1q21, 4q31, 6q21, and 8q24 (in four cases each). Numerical gain of chromosomes 7, 9, 12, and X and loss of chromosomes 1, 4, 6, 17, and Y, were noted in approximately 20% of cases. The minimum deleted regions encompassed 6q21-q25, 1p22-p36, 1q32-q44, 2p23-p25, 4q31-q35, 13p13-q14, and 17p11-p13. Two cases presented with a sole structural abnormality, and one contained a der(17)t(9;17)(p21;p13), which has not been reported earlier as a sole abnormality in DLBCL. Upon completely characterizing the karyotypes, we observed with interesting that in 55% of the cases, more than one BCL gene bearing regions was involved in translocations. In the remaining 45%, where only one or none of the BCL gene regions was involved in a rearrangement, we observed the loss of chromosomes 6 and/or 17 or partial deletions of 6q and/or 17p or gain of 7 and/or 12. Our findings suggest that, although BCL2 and BCL6 are most often implicated in DLBCL, the possibility of the disruptions of BCL3, BCL8, BCL9, and BCL10 as a "primary event" in DLBCL cannot be ruled out. Most often, a combination of events may be necessary for the genesis of DLBCL or progression of follicular lymphoma to DLBCL. Overall, M-FISH has enhanced our ability to provide a comprehensive karyotypic analysis, and has helped in defining the importance of BCL3, BCL8, BCL9, and BCL10 carrying breakpoints in DLBCL.     

Analysis of follicular lymphoma by dual-color fluorescence in situ hybridization

Follicular lymphoma (FL) is divided into two groups: one with the t(14;18)(q32;q21) and the other without this translocation but with other chromosomal abnormalities including t(3q27) break. The majority of FLs in Western countries have the former chromosomal changes with characteristic clinical features and low histological grades. The goal of this study was to investigate the characteristics of Korean FLs with regard to the underlying molecular defects. Sixty-one cases of FL were evaluated from two centers in Korea by immunostaining for CD10, bcl-2, and bcl-6 proteins. Fluorescence in situ hybridization was performed to detect the t(14;18) and t(3q27) break. Cases with FL grade 3 accounted for 57% of all 61 cases. The t(14;18) was detected in 43.9% of the cases studied, and the frequency was 80, 50, 34.8, and 9.1% from grade 1 through grade 3b. The t(3q27) was detected in 16.1% of cases. Cases without a t(14;18) were mainly histological grade 3 (P < 0.001) and had a tendency to have the t(3q27) and bcl-2 amplification. The incidence of t(14;18)-positive low-grade FL was found to be lower in Korea than in Western countries. The increased frequency of t(14;18)-negative grade 3 FL attributes to the lower incidence of t(14;18) in Korean FL.     

Cytogenetics and fluorescence in situ hybridization studies of diffuse large B-cell lymphoma in children and young adults

Diffuse large B-cell lymphoma (DLBCL), the most common subtype of adult non-Hodgkin lymphoma (NHL), is infrequently seen in adolescents and is rare in children. Due to the infrequency of the disease, single institution-based cytogenetic and fluorescence in situ hybridization (FISH) studies of pediatric DLBCL have not been reported so far and, hence, the possible differences in pediatric and adult DLBCL have not been evaluated. We performed cytogenetic and FISH analyses of 7 pediatric and 5 young adult DLBCL cases referred to the University of Nebraska Medical Center. Karyotypic studies revealed numeric and structural chromosome abnormalities in all cases. Loss of chromosomes 2, 3, 4, 6, 12, 15, 16, and 17, and gain of 12, 18, and X were observed in more than 20% of the cases (#10878;3 cases). Sex chromosome abnormalities and cytogenetically unidentifiable chromosomes and/or segments were observed in 80% (10/12) of the cases. Recurrent breakpoints (observed in 3 or more cases) included 14q32 (IGH) and 17p13 (TP53), which clustered in the young adult group. The breakpoints 7q36, 9p24, 13q34, and 16q24 were noted in two cases each. We performed interphase FISH studies to verify the possible rearrangements of the breakpoints that are frequently implicated in adult DLBCL. Our results confirmed that the pediatric cases did not show rearrangements of 3q27 (BCL6), 14q32 (IGH), 18q21 (BCL2), 8q24 (CMYC), and 17p13 (TP53), except for one case with IGH;BCL2 dual fusion [t(14;18)(q32;q21)] and one with a 17p13 (TP53) deletion. Although 3q27 was noted to be rearranged by conventional cytogenetics in two young adult DLBCL cases, FISH investigations verified that BCL6 was not disrupted. The t(8;14)(q24;q32) with rearranged CMYC ascertained by FISH, was observed in a single young adult DLBCL case. These results highlight a distinctly different representation of cytogenetic abnormalities in pediatric versus adult DLBCL.     

Focal follicular features in tonsillar diffuse large B-cell lymphomas: follicular lymphoma with diffuse areas or follicular colonization

Focal follicular features in diffuse large B-cell lymphomas (DLBCLs) are bound to raise the question of follicular lymphoma (FL) with diffuse areas, because the diagnosis of FL is based on the presence of follicular areas, even though focal. We report 7 cases of primary tonsillar DLBCLs with focal follicular features that presented with morphologic, immunohistochemical, and biological features distinct from those of FL. Histologically, these tumors were characterized by involvement of pericryptal follicles with adjacent dominant diffuse areas. Monomorphous large tumor cells were evenly spaced with abundant, often clear cytoplasm, and blastoid nuclei often with a delicate nuclear membrane. Importantly, residual germinal centers (GCs) were present in the form of either an intrafollicular GC remnant or an isolated GC in the midst of diffuse tumor. An extrafollicular and/or parafollicular growth pattern was also observed. Bcl-6 staining revealed a predominantly sporadic occurrence of Bcl-6(+) cells, comprising <50% of tumor cells, and none displayed diffusely dense collections (>75%) of Bcl-6(+) tumor cells characteristic of the GC or FL. Staining for CD10 was negative in 6 cases. Five of 7 patients were younger than 60, the median age of other patients with primary tonsillar DLBCL. No extratonsillar involvement was seen at 18 months after diagnosis. After chemotherapy or radiotherapy, complete remission was achieved with ease in all patients, but 2 patients who were treated with chemotherapy alone relapsed at 24 and 30 months. In conclusion, tonsillar DLBCL includes a small (10%) but distinct subgroup that warrants distinction from FL with predominant diffuse areas or de novo DLBCL. It appears that the focal follicular features in tonsillar DLBCL likely represent follicular colonization of marginal zone B-cell lymphoma, probably high-grade, if the possibility of FL is excluded.     

Turner综合征患儿标记染色体的来源研究

目的 了解Turner综合征患儿标记染色体的来源，以指导遗传咨询及治疗。方法 在染色体核型分析的基础上，对32例Turner综合征患者进行回顾性分析。对3例含有标记染色体的患儿进一步用荧光原位杂交技术研究标记染色体的来源。结果 3例含有标记染色体的Turner综合征患儿中，确定1例患儿的标记染色体来源于Y染色体，含有性别决定基因；1例来源于X染色体；另外1例未能确定其来源，该标记染色体可能来源于性染色体的其他片段或其他端着丝粒染色体。结论 Turner综合征患者的标记染色体大多来源于性染色体（X染色体、Y染色体），也可能来源于其他端着丝粒染色体。有必要同时应用X染色体和Y染色体特异性探针对Turner综合征患者进行标记染色体的荧光原位杂交分析，以明确标记染色体的来源。     

Semi-quantitative fluorescence in situ hybridization analysis indicates that the myc protein is consistently stabilized both before and after transformation of low-grade follicular center to high-grade diffuse large cell lymphoma.

We have investigated the expression of the MYC gene at both the mRNA and protein levels to determine how these parameters are related in lymphoma cells and in nonmalignant lymphoid cells. To do this we have adopted a multicolor fluorescence in situ hybridization methodology, which has allowed us to investigate the expression of different genes at the same time in the same cell. We have made use of the digital imaging capabilities of a charge-coupled device camera system to quantify the hybridization signals for the MYC gene and, by comparing these to the expression of a control gene (glyceraldehyde-3-phosphate dehydrogenase; GAPDH), have obtained relative quantitations of MYC mRNA and protein levels. In this study we have compared cells both within and outside the germinal centers in control tissues (reactive lymph nodes and tonsils) and in low-grade follicular center lymphomas, as well as cells in high-grade diffuse large cell lymphomas. The MYC/GAPDH mRNA hybridization signal ratios were calculated and found to be higher in cell populations containing a majority of malignant cells (p < 0.04). However, when the myc/GAPDH protein hybridization signal ratios were calculated, these were significantly higher in malignant cells from all lymphomas than the ratios observed in the nonmalignant cells (p < 0.0005). These observations indicate that the environment in a malignant cell may contribute to the stabilization of the myc protein, thus enabling it to function for a longer time period than in nonmalignant cells.     

Detection of a subset of CD30+ anaplastic large cell lymphoma by interphase fluorescence in situ hybridization

T/null-cell anaplastic large cell lymphoma (ALCL) is a morphologically and clinically heterogeneous group of non-Hodgkin's lymphoma; to date several morphologic variants have been described on histologic specimens. However, the cytologic features of these variants in the fine-needle aspiration (FNA) specimens have not been well evaluated. The t(2;5)(p23;q35) has been identified in a subset of T/null-ALCL and is known to be associated with a favorable prognosis. We reviewed the cytomorphologic characteristics in 24 FNA specimens of ALCL. In all cases, the diagnosis was confirmed on histologic specimens, and immunohistochemical studies for anaplastic lymphoma kinase (ALK) protein expression were performed on the aspirates. The presence of ALK breakpoints were evaluated in nine cases, using a DNA break-apart probe on chromosome 2 covering the ALK gene by fluorescence in situ hybridization (FISH) techniques. Two hundred cells per case were examined. The results were expressed as the percentage of cells containing more than two signals of chromosome 2 to the total number of cells counted. FNA sites included lymph nodes (20), lung (2), breast (1), and soft tissue (1). The median age of the patients was 56 yr (range, 17-75 yr). Twenty cases had systemic involvement; in four cases, skin was the primary site with secondary involvement of the lymph nodes. All cases were CD30(+) by immunohistochemistry; 20 were of T-cell phenotype and 4 were null cell type. The cytologic evaluation revealed typical anaplastic morphology (common type) with many "hallmark cells" in 16 (67%) cases. Other morphologic variants identified were small cell pattern in five cases, monomorphic pattern in two cases, and lymphohistiocytic pattern in one case. FISH studies showed that six (66.7%) of nine cases had at least two signals of chromosome 2, consistent with ALK breakpoints. With careful cytomorphologic evaluation in conjunction with appropriate immunohistochemical studies, a diagnosis of ALCL can be confidently made in the FNA specimens in the cellular aspirates and its morphologic variants also can be recognized. Furthermore, the FNA specimen is suitable in detecting ALK breakpoints by FISH study, permitting rapid identification of a subset of patients with ALCL, who may have a favorable prognosis. Using a commercially available probe, detection of ALK breakpoints in the FNA specimens is simple and can be a useful diagnostic adjunct in cases where distinction from other lymphomas or lymphoid lesions is morphologically difficult.     

Cytogenetic and fluorescence in situ hybridization studies in 60 patients with multiple myeloma and plasma cell leukemia

We report cytogenetic results in a series of 60 patients affected with multiple myeloma (MM) and plasma cell leukemia (PCL) and compare the results with those previously reported. In our series, a total of 41% of MM patients and 71% of PCL patients displayed chromosome abnormalities. To evaluate the clinical value of monosomy 18, we obtained fluorescence in situ hybridization results (using centromeric probe for chromosome 18) of 22 MM patients who displayed a normal karyotype. Monosomy 18 was present in 3 of 22 patients (14%). Using conventional cytogenetics, we detected monosomy 18 in one patient affected with PCL. Two of four cases with monosomy 18 followed an aggressive course, with overall survival of 1 and 9 months. The remaining two are in follow-up and remain stable. The association of monosomy 18 with IgA subtype predominance and poor prognosis was not observed in this series of MMs and PCLs. Although these results do not confirm our previous hypothesis, further observations of this group of patients (with monosomy 18) regarding malignant transformation is warranted.     

荧光原位杂交检测石蜡包埋间变性大细胞淋巴瘤病例中ALK基因转位及其意义

目的　比较荧光原位杂交 (fluorescence in situ hybridization,FISH)和免疫组织化学在检测间变性大细胞淋巴瘤 (anaplastic large cell lymphoma,AL CL )中间变性淋巴瘤激酶 (anaplastic lymphomakinase,AL K)基因转位及其融合蛋白中的作用 ,并探讨 FISH在石蜡包埋组织中的应用。方法　采用双色FISH和免疫组织化学检测 2 2例石蜡包埋 AL CL病例中 AL K基因转位及其融合蛋白。结果　通过调整组织切片的酶消化时间等优化措施 ,成功地在石蜡切片上进行了双色 FISH实验 ;FISH和免疫组织化学均在 6 0 % (12 /2 0 )系统性 AL CL中检测到 AL K基因转位或融合蛋白 ,在 2例皮肤原发 AL CL中未检测到基因转位或融合蛋白 ,两种方法的符合率为 10 0 %。结论　 (1)在检测 AL CL中有无 AL K基因转位时 ,AL K蛋白免疫组化由于其简单、快捷、价廉成为一般情况下的首选方法 ;在具备 FISH条件时 ,也可以将 FISH作为首选 ;(2 )通过优化实验条件 ,可以在石蜡包埋组织上成功地进行 FISH实验。     

Supernumerary ring chromosome 20 characterized by fluorescence in situ hybridization

We report on a boy with mild dysmorphic features and developmental delay, in whom karyotyping showed an additional minute ring chromosome in 60% of metaphases. Fluorescence in situ hybridization (FISH) with a centromere specific probe demonstrated that the ring chromosome contained the centromeric region of chromosome 20. The ring was highlighted completely using a chromosome 20 painting probe. A cosmid probe for 20p12-13 gave a positive signal and hybridization with an all-telomere probe showed one signal, suggesting a breakpoint in the 20p telomere. The results suggested that only a small part of 20q was involved in this ring. The ring was also detected in 18% of nuclei of a buccal smear. The phenotypic similarities of symptoms in the proband to patients with a (partial) trisomy 20p and the dissimilarities to symptoms in patients with (partial) trisomy 20q were in agreement with the FISH results.     

Chromosomal organization of amplified chromosome 12 sequences in mesenchymal tumors detected by fluorescence in situ hybridization

The chromosomal organization of amplified chromosome 12 sequences was studied with fluorescence in situ hybridization in six mesenchymal tumors: two osteosarcomas, one lipoma, two liposarcomas, and one fibrosarcoma. All except the fibrosarcoma contained ring and/or giant marker chromosomes. Amplification of chromosome 12 sequences, demonstrated with whole-chromosome paint in all cases, was confined to ring and giant marker chromosomes in four tumors. In one of the osteosarcomas and in the fibrosarcoma, amplified sequences were added to chromosome 12 and to chromosomes 10, 12, 18, and the Y chromosome, respectively. Hybridizations with single-copy probes demonstrated considerable inter- and intracellular variation in the arrangement of chromosome 12 sequences in ring and marker chromosomes. Amplification of 12q13–15 sequences, predominantly from the HMGIC–MDM2 region, was detected in all cases, but the two osteosarcomas also contained amplification of 12p material. This finding, combined with results from previous studies, indicates that 12p amplification is a feature distinguishing osteosarcomas from adipose tissue tumors. A novel finding was the presence of positive signals for chromosome 12 alpha-satellite sequences in ring and marker chromosomes in four cases. Rod chromosomes carrying amplified material, in particular those that were relatively stable, frequently exhibited chromosome 12 negative terminal segments; two of these, present in two separate cases, were shown by C-banding to contain constitutive heterochromatin. The significant intercellular heterogeneity in the number and structure of rings and giant markers in a subset of mesenchymal tumors could be explained by continuous recombination through breakage–fusion–bridge cycles. If so, this process will continue until broken ends become stabilized, for example by acquisition of telomeric segments from other chromosomes. Genes Chromosomes Cancer 23:203–212, 1998. © 1998 Wiley-Liss, Inc.     

Combination of Hodgkin's disease and diffuse large cell lymphoma: an in situ hybridization study for immunoglobulin light chain messenger RNA

It is not clear whether the rare combination of Hodgkin's disease with non-Hodgkin lymphomas are true composite lymphomas or differentiation stages of one tumour cell clone. We used in situ hybridization and immunohistochemistry for the demonstration of immunoglobulin light chains in order to investigate the relationship between the two lymphoma components. In three cases of nodular lymphocyte predominance Hodgkin's disease combined with diffuse large B-cell lymphoma the Hodgkin cells, as well as the tumour cells in the diffuse large B-cell lymphoma, showed the same messenger RNA for one light chain. Thus, using in situ hybridization in nodular lymphocyte predominance Hodgkin's disease combined with diffuse large B-cell lymphoma in a small number of cases a possible genetic relationship between the two components could be shown. In nodular sclerosis combined with diffuse large B-cell lymphoma, in situ hybridization did not support a common clonal origin of both tumour parts. However, a unique clonal derivation cannot be excluded by the techniques applied.     

Analysis of chromosome 12 aneuploidy in interphase cells from human male germ cell tumors by fluorescence in situ hybridization.

The i(12p) marker chromosome has been found to be a highly nonrandom chromosome abnormality associated with germ cell tumors (GCTs). We have previously shown that a chromosome 12 centromere specific alpha-satellite DNA probe detects the i(12p) by virtue of differences in the size of the signal originating from the i(12p) and normal chromosome 12 centromeres after fluorescence in situ hybridization (FISH) in metaphase and interphase cells of cultured GCT cell lines. We have now extended this analysis to 72 fresh GCT tumor biopsy specimens. Banded cytogenetic analysis was attempted on each of these tumors, 45 of which were found to be clonally abnormal. Data on i(12p) and chromosome 12 copy number obtained by FISH agreed well with those obtained by cytogenetic analysis. In addition, the FISH method made possible the detection and determination of i(12p) and the chromosome 12 copy number in cases in which conventional cytogenetic analysis was unsuccessful. We found the incidence of i(12p) in seminomas to be low (7%) compared to that in nonseminomas (75%) when tumor biopsy specimens were studied by FISH. Our results show that the FISH technique can be used reliably for detection of the diagnostically and prognostically useful i(12p) marker in GCT tumor biopsy specimens.     

未培养羊水荧光原位杂交快速检测60例胎儿常见染色体数目异常

目的 应用细菌人工染色体(bacterial artificial chromosome,BAC)克隆自行制备荧光探针,对胎儿常见染色体数目异常(13,18,21,X,Y)行快速产前诊断.方法 利用中国医学遗传学国家重点实验室BAC库中相应染色体特异位点克隆,自行制备荧光探针.经外周血淋巴细胞染色体杂交验证后,用于胎儿未培养羊水的快速荧光原位杂交fluorescence in situ hybridization,FISH)检测,已检测60例羊水标本.结果 所有探针特异性均为100%,杂交成功率为97.86%;FISH结果与常规核型分析一致,检出21三体2例,18三体1例.有两例涉及其它染色体的结构异常未能检出.结论 自制探针用于未培养羊水快速FISH,使用标本量少、快速、简便,可有效检出上述5种染色体的数目异常,但本法不能检出其他染色体的数目异常和结构异常,其应用仍有一定局限性.     

荧光原位杂交技术在纵隔原发生殖细胞肿瘤病理诊断中的价值

目的应用荧光原位杂交(fluorescence in situ hybridization,FISH)技术检测纵隔原发生殖细胞肿瘤(primary mediastinal germ cell tumors,PMGCTs)中12p获得的情况,探讨其在PMGCTs病理诊断中的价值。方法观察3例PMGCTs的临床病理特征,采用免疫组化En Vision法染色检测PMGCTs的免疫表型,应用FISH法检测肿瘤的12p获得情况。结果 3例PMGCTs均为男性,年龄22~24岁。CT示前纵隔占位,例3肿瘤广泛累及肺部。患者血清AFP和(或)β-hCG升高。2例肿瘤组织形态表现为单一精原细胞瘤或混合性生殖细胞肿瘤。免疫表型:SALL4、PLAP均阳性,其中精原细胞瘤成分表达CD117、OCT4,卵黄囊瘤表达AFP,胚胎癌表达CKpan、CD30。例3肿瘤分化差,免疫抗原显著丢失。FISH检测3例PMGCTs均存在i(12p)信号。结论青春期后非纯畸胎瘤PMGCTs发生12p获得,FISH检测纵隔肿瘤存在i(12p),具有重要的病理诊断价值。     

9号环状染色体综合征的荧光原位杂交分析

目的 对1例9号环状染色体综合征患儿进行细胞分子遗传学分析,探索9号环状染色体与临床表型的关系.方法 采用染色体G显带核型分析和TelVision 9p探针和TelVision 9q探针进行双色荧光原位杂交,识别和定位1例9号环状染色体患儿.结果 患儿核型为45,X,-9/46,XX,r(9)(p24q34)/46,XX,r(9;9)(p24q34;p24q34)(4/92/4).双色荧光原位杂交显示9号环状染色体上没有杂交信号,提示9号环状染色体短臂末端缺失片段至少有115 kb,长臂末端缺失片段至少有95 kb.与其它报道的环状9号染色体综合征、9号染色体短臂和长臂部分单体综合征相比,本例患者兼有环状9号染色体综合征的临床特征以及9号染色体短臂和长臂部分单体综合征的一些特征.结论 由于缺失的断裂点之间亚显微结构的不同、环的不稳定性、基因与表型相互作用以及胎儿环境条件的不同等原因,具有相同断裂点的9号环状染色体综合征患者可以有不同的临床表型,单倍基因剂量不足对临床表型发挥了重大作用.     

Identification of multiple chromosome 12 abnormalities in human testicular germ cell tumors by two-color fluorescence in situ hybridization (FISH)

The distribution of segments of the short and long arms of chromosome 12 was distinguished by two-color fluorescence in situ hybridization (FISH) in 27 cytogenetically abnormal testicular germ cell tumors (TGCTs). A 12p-specific probe was developed by chromosomal microdissection and sequence-independent polymerase chain reaction (PCR) amplification and was combined with a commercially available whole-chromosome 12 painting probe. The TGCTs included both i(12p)-positive and i(12p)-negative primary tumors and lymph node metastases from patients in clinical stage I or stage II who were not previously treated with chemotherapy. Rearrangements of the short arm of chromosome 12 and overrepresentation of 12p DNA sequences were found in all cases. In addition, cryptic rearrangements of 12p were found in 39% (7/18) of the i(12p)-positive tumors and in 78% (7/9) of the i(12p)-negative tumors. Only 7% (2/27) of all tumors had cryptic rearrangements of 12q.     

Novel chromosome findings in bladder cancer cell lines detected with multiplex fluorescence in situ hybridization

Bladder cancer is a common neoplasm worldwide, consisting mainly of transitional cell carcinomas, while squamous, adenocarcinoma, and sarcomatoid bladder cancers account for the remaining cases. In the present study, multiplex fluorescence in situ hybridization (M-FISH) has been used to characterize chromosome rearrangements in eight transitional and one squamous cell carcinoma cell line, RT112, of UMUC-3, 5637, CAT(wil), FGEN, EJ28, J82, 253J, and SCaBER. Alterations of chromosome 9 are the most frequent cytogenetic and molecular findings in transitional cell carcinomas of all grades and stages, while changes of chromosomes 3, 4, 8, 9, 11, 14, and 17 are also frequently observed. In the present study, alterations previously described, including del(8)(p10), del(9)(p10), del(17)(p10), and overrepresentation of chromosome 20, as well as several novel findings, were observed. These novel findings were a del(15)(q15) and isochromosome 14q, both occurring in three of nine cell lines examined. These abnormalities may reflect changes in bladder tumor biology. M-FISH represents an effective preliminary screening tool for the characterization of complex tumor karyotypes.     

双探针显色和荧光原位杂交法检测乳腺癌HER2基因状态和17号染色体的分析

目的 通过与荧光原位杂交(FISH)比较,评估双探针显色原位杂交(双探针CISH)法在诊断女性乳腺浸润性导管癌HER2基因状态中应用的可靠性,同时探讨17号染色体多倍体对原位杂交诊断HER2基因状态时可能产生的影响.方法 收集146例乳腺癌组织的常规石蜡包埋标本,分别用欧盟(CE)认证的FISH(146例)及CISH(73例)HER2/17号染色体着丝粒(CENl7)双探针试剂盒技术,对肿瘤标本的HER2基因状态进行检测,并按照2007年美国临床肿瘤协会及美国病理家协会(ASCO/CAP)标准分别用计算HER2基因拷贝数和(或)计算HER2/CENl7比值的方法对结果进行分析.结果 在同时进行FISH和双探针CISH检测的73例标本中发现,两种技术对HER2阴性和阳性的诊断符合率分别为91.7%(33/36)和97.4%(37/38),两者的总体符合率为95.9%(70/73);在对146例FISH结果的计数分析中,计数HER2基因拷贝数得出不确定诊断的病例量是计数HER2/CENl7得出不确定诊断病例量的1.6倍(13/8);此外,在FISH和CISH结果中按拷贝数计数为阳性的病例与在按比值计数时却为阴性病例的不吻合率分别为4.8%(3/63)和3.O%(1/33);同时在FISH HER2阳性病例(HER2/CENl7)中多倍体发生率为63.5%(40/63,P=0.002),高于阴性者中多倍体的发生率(37.3%,28/75).结论 双探针CISH技术检测乳腺癌HER2基因状态的结果和FISH技术检测的结果非常一致,提示两种技术临床诊断价值相近,并且同步检测并计数HER2和17号染色体有助于明确HER2基因状态.     

Duplication within chromosome 5q characterized by fluorescence in situ hybridization

We present a 16-month-old boy with developmental delay, minor anomalies, small penis, and lymphedema of the upper limbs. Routine cytogenetic analysis suspected a duplication of 5q. Fluorescent in situ hybridization (FISH) with a cosmid probe (MCC at the 5q22 APC region) showed tandemly duplicated fluorescent signals on one of chromosomes 5, whereas FISH with three YAC probes (TYAC12 at 5q35, HTY3182 at 5q34, and TYAC139 at 5q31) did not give duplicated signals. These findings indicate a duplication of 5q22 band in one chromosome 5. The boy we describe here is the first case of a pure partial duplication of 5q to be proven by FISH techniques. A review of previously reported cases of putative partial 5q duplication showed no consistent phenotype.