Six families with balanced chromosome translocation associated with reproductive risks in Hainan Province: Case reports and review of the literature

BACKGROUND Balanced translocation refers to the process where breakage and reconnection of chromosomes occur at abnormal positions.As the genetic substance with balanced translocation in individuals does not change,which is usually characterized by normal phenotype and intelligence,the individuals seek medical service after many miscarriages,resulting in considerable mental and physical burdens of the family members.In the current era with rapid advances in detection technology,cytogenetic examination,as a definitive approach,still plays an essential role.CASE SUMMARY We report six cases with balanced chromosome translocation:Case 1:46,XY,t(3;12)(q27;q24.1),infertility after 3 years of marriage;Case 2:46,XX,t(4;16)(q31;q12),small uterus and irregular menstruation;Case 3:46,XY,t(4;5)(q33;q13),9qh+,not pregnant after arrested fetal development;Case 4:46,XX,t(11;17)(q13;p11.2),not pregnant after two times of spontaneous abortion;Case 5:46,XX,t(10;13)(q24;q21.2),not pregnant after arrested fetal development for once;Case 6:46,XX,t(1;4)(p36.1;q31.1),not pregnant after arrested fetal development for two times.The first four cases had chromosomal aberration karyotypes.CONCLUSION These results suggested that balanced chromosomal translocation carriers are associated with reproductive risks and a very high probability of abnormal pregnancy.The discovery of the first four reported chromosomal aberration karyotypes provides an important basis for studying the occurrence of genetic diseases.     

两例急性髓系白血病伴t(6;21;8)(p22;q22;q22)复杂易位患者的临床与实验室研究

目的探讨2例急性髓系白血病（AML）伴t（6;21;8）（p22;q22;q22）复杂易位患者的临床及实验室特点.方法骨髓细胞经短期24 h培养后按常规方法制备染色体标本,R显带进行核型分析;双色双融合AML1/ETO探针进行丝裂间期及中期荧光原位杂交（FISH）检测AML1/ETO融合信号;逆转录-聚合酶链反应（RT-PCR）检测AML1/ETO融合基因转录本;综合分析临床特征.结果2例患者常规细胞遗传学分析显示均存在t（6;21;8）（p22;q22;q22）,间期和中期FISH证实了核型结果;RT-PCR检测到AML1/ETO融合基因转录本;尽管2例患者均诊断为AML-M2,但二者的免疫表型和治疗反应不同.结论t（6;21;8）（p22;q22;q22）是一种少见的t（8;21）（q22;q22）的复杂变异易位,还需要更多的病例以明确其临床特征和预后价值.     

Evidence by spectral karyotyping that 8q11.2 is nonrandomly involved in lipoblastoma

We report two cases of lipoblastoma with chromosome 8-related aberrations, ie, a 92,XXYY,t(7;8)(p22;q11.2)x2 [8]/46,XY[16] in Case 1 and a 46,XY,−8,−13,add(16)(q22),+mar, +r [cp13]/46,XY[7] in Case 2. Using spectral karyotyping and fluorescence in situ hybridization techniques, the karyotype of Case 2 was redesignated as 46,XY, r(8), del(13)(q12), der(16)ins(16;8)(q22;q24q11.2)[cp13]/46,XY[7]. This report delineates a new chromosome rearrangement, ie, der(16)ins(16;8)(q22;q24q11.2) in lipoblastoma, and also confirms the t(7;8)(p22;q11.2), reported only once previously, as a recurrent translocation involved in such a tumor. These findings provide valuable information for clinical molecular cytogenetic diagnosis of lipoblastoma. Furthermore, this report highlights the value of cytogenetic and molecular cytogenetic analysis in differential diagnosis of childhood adipose tissue tumors and adds to the number of lipoblastomas reported with chromosomal abnormalities at 8q11.2.     

Detection of translocations diagnostic for Berkitt's lymphoma by fluorescent in situ hybridization on histological sections of paraffin blocks

AIM: To test feasibility of detection of translocations which are diagnostic for Berkitt's lymphoma with the method of fluorescence in situ hibridization (FISH) on histological sections of paraffin blocks. MATERIAL AND METHODS: FISH on histological sections for detection of t(8;14)(q24;q32) and variant t(2;8)(p12;q24) and t(8;22)(q24;q11) was performed on the material obtained from 53 patients with typical clinical, morphological and immunological picture. DNA probe LSI IgH/MYC, CEP 8 Tri-color, Dual Fusion Translocation Probe (Vysis, USA), for variant translocations DNA-probe LSI MYC, Dual color, Break Apart Rearrangement Probe (Vysis, USA) were used. RESULTS: Histological material from 31 patients contained translocations characteristic for LB: in 29 (93.5%)--t(8;14)(q24;q32), in 2--variant rearrangements of locus of gene c-myc. Translocation t(8;14)(q24;q32) and its variants were not detected in 22 patients, the diagnosis was changed for diffuse large B-cell lymphoma (DLBCL). CONCLUSION: Typical for BL clinical, morphological and immunological picture may present in extra-nodal diffuse large B-cell lymphoma with high proliferative activity. Differential diagnosis between BL and the latter lymphoma is possible only basing on detection of translocation t(8;14)(q24;q32) or its variants. If it is impossible to obtain native material, FISH on histological sections of parasffin blocks is the only possible method of differential diagnosis.     

急性混合细胞白血病伴t(12;22)一例报告--附文献复习

目的报道1例伴有t（12;22）（p13;q12）的急性混合细胞白血病.方法骨髓细胞经24h短期培养后按常规方法制备染色体,采用R显带技术进行细胞遗传学分析.应用抗生物素蛋白生物素复合物（ABC）法和单克隆抗体检测白血病细胞的表面抗原;12号和22号全染色体涂染探针分别以绿色和红色2种荧光素标记后进行双色荧光原位杂交（FISH）涂染检测.结果患者的临床表现和实验室检查均符合急性混合细胞白血病.免疫表型分析髓系和淋系标记均呈阳性;染色体核型为46,XX,t（12;22）（p13;q12）[6]/46,XX,idem,der（2）[2]/46,XX[12].骨髓中期细胞经双色FISH证实12号染色体短臂和22号染色体长臂之间发生了易位.结论t（12;22）（p13;q11-13）是恶性血液病中少见的染色体异常.t（12;22）患者具有独特的临床、细胞遗传学和分子生物学特点.该染色体异常对急性白血病的预后判断价值仍需进一步观察.     

Chromosomal abnormalities in myelodysplastic syndromes and acute myeloid leukemia

Clonal chromosome abnormalities are found in more than half the patients with hematologic malignancies. Karyotype is an independent prognostic factor in these patients. Cytogenetic findings correlate significantly with morphologic, immunologic, and clinical features as well as response to treatment, remission duration, and survival. The number of different cytogenetic abnormalities is enormous; however, many cytogenetic findings frequently occur in a given disease (e.g., abnormalities of 5 or 7 in 75% to 90% of patients with therapy-related AML). Some abnormalities are found only in myeloid malignancies, for example, the t(8;21)(q22;q22) and rearrangements of chromosome 16q22, both of which have a good prognosis. Other abnormalities usually are found in both myeloid and lymphoid malignancies, for example, the t(4;11)(q21;q23) and t(9;22)(q34;q11), both of which have a poor prognosis. The Human Gene Mapping Conferences have compiled much cytogenetic data and produced several interesting correlations in myeloid malignancies: rearrangements of 3q21-26 with myeloid proliferations associated with environmental exposure (similar to abnormalities of 5q, 7q, 12p, and 17q), aberrations of 12p, 11q13 and 11q23 with both myeloid and lymphoid disorders, and the lack of myeloid involvement and abnormalities of chromosomes 14 and 18. In conclusion, cytogenetic analysis of neoplastic cells at diagnosis for patients with MDS, AML, and SAML is required for appropriate diagnosis and treatment. The use of chromosome abnormalities to separate patients into high- and low-risk groups eventually may allow us to be more effective in selecting curative therapy.     

Simultaneous translocation of both TCR Loci (14q11) with rare partner loci (Xq22 and 12p13) in a case of T-lymphoblastic leukemia

The most common recurrent cytogenetic abnormalities in T-lymphoblastic leukemia (T-acute lymphoblastic leukemia [T-ALL]) involve T-cell receptor (TCR) loci and a variety of partner genes, including HOX11, HOX11L2, MYC, and TAL1. In this report, we present a rare case involving simultaneous translocation of the TCR α/δ loci with different partner loci (Xq22 and 12p13); this resulted in a poor prognosis. Chromosomal analysis showed 46,Y,t(X;14)(q22;q11.2),t(12;14)(p13;q11.2) and FISH analysis by using a T-cell receptor alpha delta DNA probe, Split Signal (DakoCytomation, Denmark), showed translocations at the same TCR α/δ locus on both chromosomes. FISH with 2 bacterial artificial chromosome clones showed break apart signal, which suggests involvement of the IRS4 gene. To our knowledge, this is the first report of T-ALL in which both TCR α/δ loci were translocated with different partner loci, and 1 of the partner loci, Xq22, was a rare translocation partner locus that included IRS4 gene.     

CD5-negative blastoid variant mantle cell lymphoma with complex CCND1/IGH and MYC aberrations

The coexistence of CCND1/IGH and MYC rearrangements in mantle cell lymphoma (MCL) is a rare finding associated with a very poor prognosis. In this study, a patient with blastoid variant (MCL) is reported. The disease was clinically aggressive and refractory to chemotherapy, and the patient only survived for 1 month following diagnosis. Conventional cytogenetic study, FISH, and multicolor FISH (mFISH) demonstrated the involvement of the BCL1/CCND1 locus in a complex translocation, t(3;11)(q25;p15)t(11;14)(q13;q32). In addition, subclonal abnormalities in the 8q24 region, manifested as a t(8;14)(q24;q32)/MYC rearrangement, were identified. To the best of our knowledge, this is the first MCL case in Korea bearing these complex genomic aberrations.     

Cytogenetics of mantle cell lymphoma

AIM: To determine the type and rate of secondary chromosomal aberrations and role in pathogenesis of mantle cell lymphoma (MCL). MATERIAL AND METHODS: Standard cytogenetic examination (SCE) and fluorescent in situ hybridization (FISH) with tests for 11q22/ATM, 13q14, 17p13/p53, 9p21/p16 and chromosome 12 centromere were made in 28 patients. RESULTS: Secondary chromosomal aberrations were detected in 22 patients. Chromosomal abnormalities occurring in more than 2 cases include deletions 6q15-q23 (53% cases); 11q22/ATM (50%); 9p21 (36%, in half the cases deletion 9p21 was biallele); 13q14 (32%); trisomy 3/3q, monosomy 20 and deletions/translocations 1q and 1p (27% and 20%) and deletion 17p13/p53 (18%). Trisomies 12/12q, 6 and 18/18q, monosomies 2 and 16, deletions/translocations 2p and 16p were found in 2 cases each. The FISH technique identified 9 chromosomal anomalies missed at SCE. Deletions 6q15-q23 were seen in patients with privalent lesions of lymph nodes. Deletions 11q, 13q14, 9p21 and 17p13 occur more frequently in transformation and the blastoid variant. Monosomy 20 was found only in patients with large cell transformations. This was the only cytogenetic defect characteristic for transformed cases, in contrast to denovoblastoid ones. Thus, deletions 6q, 11q23, 13q14 and 9p21 are typical for MCL. Deletions 9p21 and 17p13 are more characteristic for large cell variants of the tumor. Deletion 17p13 occurs at the terminal stage of the disease in rapidly growing tumor mass and resistance to chemotherapy. FISH technique is effective in detection of submicroscopic rearrangements especially small deletions. No significant differences were found between transformed and de novo blastoid cases. This shows that the blastoid variant is not a specific biological form, it is rather a less typical manifestation of the disease due to some preclinical cytogenetic disorders. CONCLUSION: Progression and transformation of MCL are related to mutation of proapoptotic genes and genes proteins of which are inhibitors of active complexes of D1 cycline. Specification of these mechanisms requires further investigations in patients with different clinicomorphological forms of the disease at various stages of the disease.     

Prognosis and treatment outcome in patients with acute myeloid Leukemia with t(8;21)(q22;q22)

Patients with acute myeloid leukemia (AML) with the t(8;21) karyotype generally have a favorable clinical course, but key prognostic factors remain poorly defined. This study was conducted to determine the prognoses and treatment outcomes of patients with AML with this unique cytogenetic change. A total of 22 patients with AML with t(8;21)(q22;q22) were studied. Various parameters were tested for their impact on disease-free survival (DFS) and overall survival (OS). Another 55 patients with AML with a normal karyotype were included for comparison of clinical outcomes. Between patients with t(8;21) and those with a normal karyotype, no significant differences were noted in DFS (median survival, 15.23 vs 12.03 mo;P=.7626) and OS (median survival, 19.17 vs 18.93 mo;P=.7543). Among t(8;21)(q22;q22) patients, no clinical parameters showed a significant impact on DFS. Univariate analysis revealed that a higher platelet count (> 15·10⁹/L) at diagnosis, a low white blood cell count (index ≤20), and hematopoietic stem cell transplantation (HSCT) as postremission therapy were associated with improved OS. On multivariate analysis, HSCT as postremission therapy and white blood cell count index < 20 remained good independent prognostic factors for OS. The data presented here suggest that t(8;21)(q22;q22) cytogenetic changes in patients with AML had prognostic significance similar to that in patients with a normal karyotype; patients who harbored either karyotype had parallel clinical outcomes. It is concluded that patients with AML with t(8;21)(q22;q22) would be compromised by treatment approaches that do not include HSCT as postremission therapy.     

55例急性早幼粒细胞白血病形态学、细胞免疫学、细胞遗传学及分子生物学(MICM)分型的回顾性分析

为了解在已经确诊的急性早幼粒细胞白血病(APL)患者中骨髓细胞形态学(M)、细胞免疫学(Ⅰ)、细胞遗传学(C)和分子生物学(M)4者即MICM的关系及对临床诊断的指导意义，对55例APL患者的MICM分型检测结果进行了回顾性总结分析。结果显示，以FAB分型为基础的形态学确诊率可达96．4％；免疫表型检测以CD33和CD13阳性共表达率为最高，达96．4％；APL特征性染色体异常检出率为87．3％，其中核型为t(15；17)(q22；q21)100％易位者(单纯型)占75％，其它可累及的染色体有1，8，9，11，12，21号；PML/RARα基因检测阳性率达96．4％。但MICM联合检测用于APL诊断的准确率可达100％。结论：骨髓细胞形态学观察仍然是APL确诊的基础，MICM联合应用可显著提高APL的确诊率，减少误诊率；MICM联合检测，可能为发现APL新的亚型提供线索。     

Interphase cytogenetics for the detection of the t(11;22)(q24;q12) in small round cell tumors.

Among the small round cell tumors differential diagnosis is particularly difficult for their undifferentiated or primitive character. In this mixed group of tumors, only the primitive neuroectodermal tumors, which include Ewing's sarcoma (ES), show the unique and consistent feature of the (11;22)(q24;q12) translocation, which can therefore be considered a hallmark of these neoplasias. We analyzed four primitive neuroectodermal tumor cell lines, one osteosarcoma cell line, and 11 patients by fluorescent in situ hybridization with cosmid clones 23.2 and 5.8, bracketing the t(11;22) at 11q24. Metaphase spreads from tumor cell lines, and from biopsy specimens of three patients with ES were analyzed. In the remaining eight patients comprising five ES, two small cell osteosarcomas and one chronic osteomyelitis, only nuclei preparations were available for analysis. We detected the t(11;22) in interphase nuclei of the four primitive neuroectodermal tumor cell lines, of three patients in which the karyotype demonstrated the translocation and in five cases of ES in which cytogenetic analysis had not been possible. Two cases of small cell osteosarcoma and one chronic osteomyelitis were also analyzed and were both normal with respect to the t(11;22). By analyzing cell lines and small round cell tumor samples by fluorescent in situ hybridization, we established that interphase cytogenetics is a rapid alternative to chromosomal analysis for the detection of the t(11;22) and represents an invaluable tool for the differential diagnosis of small round cell tumors.     

干扰素-α治疗Ph+慢性粒细胞白血病的细胞遗传学反应分析

为了研究慢性粒细胞白血病 (CML)慢性期患者应用干扰素 α (IFN α)治疗后细胞遗传学疗效及其影响预后的因素 ,对我院 10年来 12 8例CML慢性期患者单用IFN α或联用化疗药物后细胞遗传学变化、核型演变及与临床有关特征的关系进行了回顾性分析。核型分析全部应用G 显带 ,部分联合应用荧光染色体原位杂交技术检测。结果表明 :①所有患者均获血液学缓解。② 118例Ph染色体标准易位患者中 36例 (30 .8% )获细胞遗传学反应 ,其中 2 0例 (17.1% )Ph染色体仍 >35 % ,13例 (11.1% )Ph染色体 <35 % ,3例 (2 5 % )Ph染色体为 0 ,达完全细胞遗传学缓解 ,细胞遗传学有效应者共 16例 (13.6 % )。③ 7例复杂变异易位患者中 4例获细胞遗传学反应 ,其中2例 (14 .3% )Ph染色体 >35 % ,2例 (14 .3% )Ph染色体 <35 % ,无 1例Ph染色体为 0 ;3例简单变异易位患者无 1例获细胞遗传学疗效。④IFN α治疗后影响细胞遗传学疗效的因素有 :性别、初诊病情、IFN α是否联用其他化疗药物及是否持续治疗。⑤IFN α治疗并不能防止CML疾病进展。结论 :①每周IFN α 6 0 0 - 90 0万U单用或联合Bu/Hu可使 11.1%的标准易位和少数复杂变异易位Ph+ CML患者获主要细胞遗传学效应 ,但不能防止疾病进展 ;②Ph变异易位并不预示IFN α疗效不佳 ;     

系统性轻链型淀粉样变性细胞遗传学异常检测分析

目的研究系统性轻链型(AL)淀粉样变性患者的细胞遗传学特征。方法收集初诊系统性AL淀粉样变性患者24例和多发性骨髓瘤(MM)患者135例,通过CD138磁珠分选(MACS)结合间期荧光原位杂交(FISH)技术检测系统性AL淀粉样变性和MM患者的细胞遗传学异常,比较二者细胞遗传学异常差异。分析系统性AL淀粉样变性FISH异常与血清游离轻链(sFLC)、N端脑钠肽前体(NT-proBNP)、B型脑钠肽前体(proBNP)、血清肌钙蛋白(cTnT和cTnI)及器官累及之间的关系。结果系统性AL淀粉样变性细胞遗传学异常阳性率为62.5%,其中14q32异位、t(11;14)和+1q21发生率较高,分别为41.7%、37.5%和29.2%。系统性AL淀粉样变性细胞遗传学异常总阳性率、del(13/13q14)缺失率及并存大于或等于3种遗传学异常的发生率均显著低于MM患者,差异有统计学意义(P0.05),而t(11;14)发生率显著高于MM,差异有统计学意义(P0.05)。系统性AL淀粉样变性细胞遗传学异常与临床相关检测指标及器官累及无显著关系(P0.05)。结论 MACS-FISH可用于检测系统性AL淀粉样变性患者的细胞遗传学异常。系统性AL淀粉样变性患者14q32异位、t(11;14)、+1q21有较高的发生率,t(11;14)发生率显著高于MM患者,而并存大于或等于3种细胞遗传学异常和del(13/13q14)发生率显著低于MM患者。     

Fluorescence in situ hybridization analysis of immunoglobulin heavy chain translocations in plasma cell myeloma using intact paraffin sections and simultaneous CD138 immunofluorescence

Fluorescence in situ hybridization (FISH) studies are much more sensitive than classical cytogenetics for identification of karyotypic abnormalities in plasma cell myeloma. However, FISH analysis of bone marrow samples is often challenging because of a large number of admixed non-neoplastic hematopoietic elements. In this report, we describe a novel method using FISH analysis of intact paraffin sections of formalin-fixed, bone marrow clot preparations with simultaneous CD138 tyramine signal amplification (TSA)-mediated immunofluorescence. We studied 22 cases of plasma cell myeloma for translocations involving the immunoglobulin heavy chain locus that are of known diagnostic and/or prognostic significance. All cases were analyzed using dual color, break-apart immunoglobulin heavy chain probe and dual color, dual fusion probes for t(11;14)(q13;q32) and t(4;14)(p16;q32). TSA-mediated fluorochrome deposition in CD138+ cells was unaltered by protease pretreatment. Translocations were identified in 10 cases, including five with t(11;14)(q13;q32) and three with t(4;14)(p16.3;q32). When present, abnormalities were identified in a large percentage of CD138+ cells (47 to 93%, median 84%). This technique allows for efficient molecular cytogenetic analysis of plasma cell myeloma using routinely archived paraffin-embedded material.     

119例慢性髓细胞白血病急变期核型分析

为了探讨慢性髓细胞白血病急变期（CML-BC）核型变化规律及意义，应用R显带技术对119例CML-BC患者的核型资料进行分析，并在其中随机选出28例以荧光原位杂交法（FlSH）检测衍生9号染色体[der（9）]是否缺失．结果表明：124例核型检查中Ph阴性11例（8．9％），P11阳性113例（91．1％），其中典型易位104例（83．9％），单纯变异易位4例（3．2％），复杂变异易位5例（4．0％）；Ph阴性CML-BC中72．6％伴有附加染色体异常，主要类型是：i（17q）、＋14，Ph阳性者72．3％伴有附加染色体异常，主要类型是：＋Ph、＋8、i（17q）；Ph阴性CML-BC急变时间为2-66月，平均29．0月，Ph阳性者急变时间为1-127月，平均34．2月，两组比较无统计学差异（P〉0．05）；FlSH检测发现5例（5／28，17．9％）der（9）缺失。结论：慢性髓细胞白血病急变期附加染色体异常多见，FlSH技术可有效检测der（9）缺失．