多发性骨髓瘤中14q32易位与13q14缺失的相关性研究

目的 研究多发性骨髓瘤(MM)常见的分子遗传学异常14q32易位与13q14缺失及其与临床指标的关系.方法 采用间期荧光原位杂交(I-FISH)技术应用RB1、D13S319和LSI IGHC/IGHV探针检测49例MM患者骨髓标本中RB1基因、13q14.3缺失及14q32易位,结合临床资料作统计分析.结果 49例MM患者有26例(53.1%)检测到14q32易位,25例(51.02%)存在13q14缺失(其中18例检测到13q14.3缺失,9例存在RB1缺失).Spearman相关分析显示,14q32易位多见于浆细胞比例高的患者(r=0.316,P=0.27),与患者年龄、国际分期系统(ISS)分期、免疫球蛋白分型、β2微球蛋白及肾损害无相关性(P＞0.05).结论 13q14缺失及14q32相关的易位在MM中发生率均较高,两者有密切相关性;14q32易位的MM患者浆细胞百分比明显升高,14q32易位的检测可作为预测MM预后的指标.     

多发性骨髓瘤中14q32易位与13q14缺失的相关性研究

目的 研究多发性骨髓瘤(MM)常见的分子遗传学异常14q32易位与13q14缺失及其与临床指标的关系.方法 采用间期荧光原位杂交(I-FISH)技术应用RB1、D13S319和LSI IGHC/IGHV探针检测49例MM患者骨髓标本中RB1基因、13q14.3缺失及14q32易位,结合临床资料作统计分析.结果 49例MM患者有26例(53.1%)检测到14q32易位,25例(51.02%)存在13q14缺失(其中18例检测到13q14.3缺失,9例存在RB1缺失).Spearman相关分析显示,14q32易位多见于浆细胞比例高的患者(r=0.316,P=0.27),与患者年龄、国际分期系统(ISS)分期、免疫球蛋白分型、β2微球蛋白及肾损害无相关性(P＞0.05).结论 13q14缺失及14q32相关的易位在MM中发生率均较高,两者有密切相关性;14q32易位的MM患者浆细胞百分比明显升高,14q32易位的检测可作为预测MM预后的指标.     

荧光原位杂交技术在多发性骨髓瘤患者del(13q14)检测中的应用

 目的　了解多发性骨髓瘤（MM）患者中13号染色体缺失情况,分析13号染色体部分缺失在MM中的临床价值。方法　采用荧光原位杂交（FISH）技术检测38例MM患者骨髓标本中Rb-1基因和13q14位点的缺失;采用Fisher确切概率法分析13号染色体部分缺失与患者确诊时临床特征间关系。结果　在38例MM患者中,13号染色体部分缺失20 例,其中仅Rb-1基因缺失4例,仅13q14位点缺失2例,两位点同时缺失14例。Fisher确切概率法分析显示13号染色体长臂缺失（del 13q14）与患者血清乳酸脱氢酶升高及国际分期系统（ISS）有关。结论　Rb-1基因和13q14位点的缺失在MM中均较为常见,13号染色体部分缺失对MM的生物学行为有一定影响,del (13q14)在 MM中的意义有待进一步探讨;FISH是一种在分析 MM患者13号染色体异常方面较为快速、准确和敏感的方法。     

应用荧光原位杂交技术检测多发性骨髓瘤

 目的　应用荧光原位杂交（FISH）技术检测多发性骨髓瘤（MM）常见基因异常。方法　采用1q21／RB1、D13S319／p53、IGH序列特异性基因探针,应用FISH技术对按照WHO诊断标准确诊的44例初诊MM患者进行检测,并与常规细胞遗传学技术（吉姆萨显带）检测结果及临床参数对比。结果　FISH技术检测44例初诊MM患者中32例（72.7 %）基因异常,其中1q21扩增11例（25.0 %）,RB1缺失17例（38.6 %）,D13S319缺失16例（36.4 %）,p53缺失6例（13.6 %）,IGH基因重排19例（43.2 %）。检出1种异常者10例（22.7 %）,同时有2种异常者11例（25.0 %）,3种异常者8例（18.2 %）,4种异常者3例（6.8 %）。44例应用常规吉姆萨显带检测16例无分裂象,28例有分裂象者中检出异常核型2例（7.14 %）,FISH技术基因异常检出率明显高于常规吉姆萨显带技术,差异有统计学意义（P＜0.05）。基因异常与临床参数比较结果显示,β2-微球蛋白（β2-MG）与IGH基因重排呈正相关（P＜0.05）,骨髓浆细胞数与D13S319缺失呈正相关（P＜0.05）,CRE与p53缺失及IGH基因重排呈正相关（P＜0.05）,CRP与p53缺失呈正相关（P＜0.05）。基因异常与MM分型、分期、年龄分组之间无明显相关性（P＞0.05）。结论　MM最常见的基因异常是IGH基因重排以及RB1和D13S319缺失,其次是1q21扩增,p53缺失最少。FISH可检测出与预后密切相关的累及基因位点的微缺失,异常克隆检出率较常规细胞遗传学技术显著提高,FISH可作为MM患者的常规检查,以评估预后,指导临床治疗。     

成套荧光原位杂交探针在慢性淋巴细胞白血病染色体异常检测中的应用

目的 探讨成套探针荧光原位杂交（FISH）技术在慢性淋巴细胞白血病（CLL）患者染色体异常检测中的应用,并初步分析染色体异常与其他预后指标的相关性及其临床意义.方法 对21例初诊CLL患者同时应用序列特异性探针D13S25（13q14.3）、RB1（13q14）、ATM（11q22.3）、p53（17p13.1）及着丝粒探针CSP12（12p11.1-12q11.1）进行间期FISH检测,分析患者染色体异常的发生率,同时分析染色体异常与CLL患者年龄、性别、Binet分期、CD38表达及乳酸脱氢酶（LDH）间的相关性,并对生存情况进行初步观察.结果 21例CLL患者中,13例发现有染色体异常（61.90％）,其中11例为单条染色体异常,1例为2条染色体异常,1例为3条染色体的复杂异常.按染色体异常发生率的高低依次为13q14-9例（42.86％）（其中D13S25单独缺失7例,D13S25和RB1同时缺失2例）,＋123例（14.29％）,11q22-（ATM缺失）2例（9.52％）,17p13-（p53缺失）2例（9.52％）.染色体异常与患者年龄、性别、Binet分期、CD38表达以及LDH水平未发现相关性.结论 成套探针FISH技术能够快速、敏感、准确地检测CLL患者的染色体异常.     

13号染色体、p53基因缺失与多发性骨髓瘤的相关性研究

 目的　分析13号染色体、p53基因缺失与多发性骨髓瘤（MM）患者临床特点、治疗效果和生存期的相关性。方法　采用荧光原位杂交（FISH）技术检测22例MM患者del 13q14（RB1基因缺失）、p53基因缺失的情况,收集其临床资料,比较基因缺失组与基因正常组间的差异。结果　22例患者中,9例（40.9 %）为p53基因缺失,其中Ⅰ期0例,Ⅱ期2例（40.0 %）,Ⅲ期7例（50.0 %）。8例（36.4 %）患者为RB1基因缺失,其中Ⅰ期0例,Ⅱ期3例（60.0 %）,Ⅲ期5例（35.7 %）。基因缺失组与基因正常组比较,患者的年龄、性别、分型、分期、肾功能、血细胞计数、C反应蛋白（CRP）、乳酸脱氢酶（LDH）、血钙均无差异。但初诊时骨髓瘤细胞比例,p53基因正常组明显高于缺失组（P＝0.043）。22例患者治疗后共获得CR 5例,非常好的部分缓解（VGPR）7例,PR 6例,无效4例,总有效率为81.8 %。p53基因缺失组与正常组平均生存时间分别为11和47个月（P＝0.086）。RB1基因缺失组和正常组平均生存时间分别为14.9和43.3个月（P＝0.612）。2个基因均缺失组与正常组相比,平均生存期分别为9.4和45.4个月（P＝0.13）。结论　细胞遗传学的异常与MM患者的生存期具有显著的相关性。对于具有与不良预后相关的染色体异常的患者,传统化疗效果较差,应给予硼替佐米等新药治疗。     

重组人核心蛋白聚糖基因增强多柔比星对白血病K562细胞抑制作用的实验研究

 目的　探讨慢性淋巴细胞白血病（CLL）的分子遗传学异常情况及其临床意义。方法　对17例初诊CLL患者进行常规细胞遗传学（CC）检测及应用着丝粒探针CSP12（12p11.1～12q11.1）和序列特异性探针D13s25（13q14.3）、ATM（11q22.3）、RB1（13q14）、p53（17p13.1）进行间期荧光原位杂交（I-FISH）检测。结果　CC检测18.75 %患者有核型异常,1例未见核分裂象;I-FISH检测70.6 %患者有分子遗传学异常,13q-异常47.1 %（RB1缺失23.5 %、D13S25缺失29.4 %）、+12异常29.4 %、p53基因缺失11.8 %、ATM缺失5.6 %、复杂基因组异常11.8 %。分子遗传学异常与性别、年龄、乳酸脱氢酶（LDH）、β2 -微球蛋白（β2-MG）及 Binet 分期无明显相关性。结论　I-FISH是检测CLL患者基因组异常的有效手段,与CC方法相比可明显提高CLL分子遗传学异常的检出率,分子遗传学异常与临床分期及其他临床指标无明显相关性,对患者预后的意义有待进一步研究。     

常规细胞遗传学和荧光原位杂交方法检测14q32/IgH基因重排

  目的 比较常规细胞遗传学（CC）,间期荧光原位杂交（FISH）技术及连续R显带后FISH检测免疫球蛋白重链（IgH）基因重排的应用价值。方法 应用常规细胞遗传学及间期FISH分析血液系统肿瘤患者43例。结果 43例患者中14q32+／IgH+患者19例,14q32-／IgH+患者2例（4.7 %）,14q32+／IgH-患者3例（7.0 %）,14q32-／IgH-患者19例。部分病例CC与间期FISH方法检测14q32／IgH基因重排得到了不一致的结果。对5例患者进行连续R显带后FISH分析,对照核型和FISH图,能清楚地看到IgH基因易位涉及的染色体。结论 间期FISH可以提高14q32／IgH重排检出率,R显带后FISH可以帮助识别与14q32易位的伙伴染色体。     

多发性骨髓瘤患者临床特征、FISH检测结果与疗效相关性的临床分析

目的:探讨初治多发性骨髓瘤（multiple myeloma,MM）患者的临床特征、FISH检测结果与不同治疗方案疗效的相关性。方法:回顾性分析54例初治多发性骨髓瘤患者临床特征、FISH检测结果、治疗方案及疗效,利用独立样本t检验比较分子遗传学与患者临床特征的关系,利用χ2检验或Fisher确切概率法比较不同治疗方案的疗效与分子遗传学异常的关系。结果:54例MM患者总有效率为61.11%（33/54）。完成FISH检测者共41例,伴分子遗传学异常者占70.73%（29/41）,其中1q21扩增、13q14缺失、p53缺失、IgH重排检出率分别为53.66%（22/41）、31.71%（13/41）、2.44%（1/41）、36.59%（15/41）,2种及以上分子遗传学异常者占48.78%（20/41）。13q14缺失阳性组患者血清校正钙水平较阴性组高（P＜0.05）,IgH重排阳性组患者骨髓浆细胞比例高于阴性组（P＜0.05）。伴有2种及以上分子遗传学异常患者总有效率及完全缓解率均低于伴有1种分子遗传学异常患者（P=0.014,P=0.005）。传统化疗组中13q14缺失、1q21扩增阳性患者总有效率均较阴性患者低（P=0.02,P=0.03）,未发现IgH重排阳性患者与阴性患者间疗效差异（P＞0.05）;硼替佐米组中基因异常患者与正常患者间总有效率相比差异无统计学意义（P＞0.05）。结论:新型治疗药物的出现使MM患者疗效得到改善,分子遗传学异常与临床特征及疗效相关,可协助判断患者预后并为治疗方案的选择提供依据,其中伴有13q14缺失阳性及1q21扩增患者治疗反应差,蛋白酶体抑制剂硼替佐米可在一定程度上改善分子遗传学阳性患者近期疗效。     

应用FISH技术检测慢性淋巴细胞白血病ATM 和RB1基因缺失*

目的:探讨慢性淋巴细胞白血病（CLL）中11q22.3（ATM基因）和13q14（RB1 基因）的缺失情况以及荧光原位杂交（fluorescence in situ hybridization,FISH）技术检测慢性淋巴细胞白血病（CLL）染色体异常的价值,了解CLL 的分子遗传学特性。方法:分别用ATM和RB1 探针,运用荧光原位杂交（FISH）技术对10例CLL 确诊患者的11q22.3 和13q14染色体进行检测,并和常规细胞遗传学（Conventional cytogenetics,CC）检测方法,即G 显带法进行比较。结果:10例CLL 患者常规细胞遗传学检出2 例,其中t（5,9）,t（10,12）和+ 12各1 例;而FISH方法检出7 例至少有一种分子遗传学异常,del（11q22.3）4 例,纯合缺失2 例,del（13q14）7例,纯合缺失2 例。10例患者中4 例同时有del（11q22.3）和del（13q14）这2 种染色体异常。结论:FISH是一种在分析CLL 染色体异常方面较为快速、准确和敏感的有效技术,可提高染色体异常检出率,为CLL 提供较为准确的分子细胞遗传学信息,指导临床与预后分析。      

Both IGH translocations and chromosome 13q deletions are early events in monoclonal gammopathy of undetermined significance and do not evolve during transition to multiple myeloma.

Molecular and genetic events associated with the transition from monoclonal gammopathy of undetermined significance (MGUS) to multiple myeloma (MM) are still poorly characterized. We investigated serial bone marrow specimens from 11 patients with MGUS who eventually progressed to MM (MM post-MGUS) by interphase fluorescence in situ hybridization for immunoglobulin heavy-chain gene (IgH) translocations and chromosome 13q deletions (del(13q)). In nine patients, IgH translocations were present both in MGUS and MM post-MGUS plasma cells, including three t(11;14)(q13;q32) and one t(4;14)(p16;q32), which was observed already 92 months prior to MM. Similarly, all five MM patients with del(13q) had this aberration already at the MGUS stage. Two patients without IgH translocation and del(13q) had chromosomal gains suggesting hyperdiploidy, but IgH translocations and/or del(13q) did not emerge at MM post-MGUS. IgH translocations and del(13q) are early genetic events in monoclonal gammopathies, suggesting that additional events are required for the transition from stable MGUS to progressive MM.     

iFISH技术检测多发性骨髓瘤遗传学异常及其临床意义

目的:采用iFISH技术了解多发性骨髓瘤染色体异常情况,分析染色体异常与多发性骨髓瘤部分临床资料、治疗效果及预后之间的关系。方法:采用组合探针（1q21、DLEU/RB1、p53、IGH）对94例初诊MM患者骨髓细胞进行iFISH检测,分析其细胞遗传学异常,探讨其与临床资料及治疗效果之间的关系,明确其预后价值。结果:94例患者中,74例（78.72%）检测出1种及1种以上细胞遗传学异常;其中1种异常的34例（34/74,45.95%）,2种异常的15例（15/74,20.27%）,25例存在3种及3种以上异常（25/74,33.78%）。其异常的比例从高到低分别为:p53缺失（45/74,60.81%）,1q21扩增（44/74,59.46%）,IGH重排（31/74,41.89%）,DLEU/RB1缺失（28/74,37.84%）。分析发现,p53缺失与血小板计数、白蛋白及球蛋白水平有关,IGH重排与白蛋白及瘤细胞负荷有关,DLEU/RB1缺失与血红蛋白、血沉、白蛋白及球蛋白水平有关,未发现1q21扩增与各项临床指标之间的关系。将患者按照治疗方案分为VAD组和硼替佐米组,结果显示VAD组中iFISH阴性者的疗效明显优于iFISH阳性者,而在硼替佐米组未发现FISH阳性者和阴性者的疗效有差异。生存分析发现,IGH重排与总生存时间相关,而p53缺失与总生存时间和无病生存时间均明显相关,为MM患者的独立预后因素。结论:多数MM患者存在细胞遗传学异常;1q21扩增、p53缺失及IGH重排的发生率较高;细胞遗传学异常与多种不良临床指标相关,并与临床治疗反应性相关,存在异常者治疗效果欠佳,预后较差。     

Deletions of chromosome 13q in monoclonal gammopathy of undetermined significance.

Since deletion of chromosome 13q is a clinically relevant feature in multiple myeloma (MM), we analyzed bone marrow plasma cells from 29 patients with monoclonal gammopathy of undetermined significance (MGUS) to investigate the chromosome 13 status in MGUS. Studies were performed by interphase fluorescence in situ hybridization (FISH) with a panel of 13q14-specific probes (RB1, D13S319, D13S25, D13S31). Plasma cells with a deletion of at least one of the 13q14 loci were detected in 13 patients (44.8%) with MGUS. In five patients (17.2%), deletions of all four 13q14-specific probes were observed, and the additional deletion of a 13q telomeric region (D13S327) suggested loss of the entire 13q arm or monosomy 13. Loss of 13q14 was observed to be monoallelic and to occur in 11.0 to 35.0% of plasma cells (cut-off levels for a deletion <10% with all probes). Nine of 17 patients (52.9%) with MM progressing from a pre-existing MGUS had evidence for a deletion of 13q14 as determined by FISH with the RB1 probe. These results suggest that deletion of 13q14 is an early event in the development of monoclonal gammopathies, but its role for the eventual progression to MM remains to be determined prospectively.     

t(11;14) and t(4;14) translocations correlated with mature lymphoplasmacytoid and immature morphology, respectively, in multiple myeloma.

Recent studies have shown that two recurrent translocations, t(4;14)(p16;q32) and t(11;14)(q13;q32), define distinct entities with different prognosis in multiple myeloma (MM). We addressed the issue of whether these illegitimate IGH rearrangements could contribute to the morphological heterogeneity of the malignant plasma cells (PC). Bone marrow aspirates of 178 untreated MM cases with successful molecular cytogenetics analysis using fluorescence in situ hybridization were reviewed. PC of 25/48 (52%) patients with t(11;14) exhibited a lymphoplasmacytoid morphology. Moreover, 25/27 (93%) of the cases with this morphological profile bore the t(11;14). In addition, both cytogenetics and morphological subtypes shared higher incidence of nonsecretory MM. In contrast, 17 out of 28 cases (61%) with t(4;14) exhibited PC with diffuse chromatin pattern. Interestingly, both t(4;14) translocation and immature morphology correlated with higher incidence of high tumor mass and chromosome 13 abnormality. In conclusion, our results suggest that a particular morphology can be the signature of chromosomal abnormalities in MM.     

14q32 translocations and monosomy 13 observed in monoclonal gammopathy of undetermined significance delineate a multistep process for the oncogenesis of multiple myeloma. Intergroupe Francophone du Myélome.

Clonal plasma cells in monoclonal gammopathy of undetermined significance (MGUS) have been shown to bear copy number chromosome changes. To extend our knowledge of MGUS to structural chromosomal abnormalities, we have performed fluorescence in situ hybridization experiments with probes directed to the 14q32 and 13q14 chromosomal regions in 100 patients with either MGUS or smoldering multiple myeloma (SMM). 14q32 abnormalities were observed in at least 46% of patients with MGUS/SMM, with these abnormalities being present in the majority of clonal plasma cells. Whereas t(11;14)(q13;q32) occurs in 15% of MGUS/SMM patients, an incidence similar to that of overt multiple myeloma (MM) patients, translocation t(4;14)(p16;q32) is observed in only 2% of these cases [P = 0.002 for difference with t(11;14)], as compared with 12% in MM patients (P = 0.013). Monoallelic deletions of the 13q14 region were found in 21% of patients, with two types of situations. In half of the evaluable patients, and especially in patients with SMM, the deletion is present in the majority of clonal plasma cells, as in MM, whereas in the other half of the evaluable patients (essentially in MGUS patients), it is observed in subclones only. These data enable us to elaborate a plasma cell oncogenesis model from MGUS to MM.     

Cytogenetic Abnormalities in Multiple Myeloma Patients at a Tertiary Healthcare Center in India

Objective: Multiple myeloma (MM) is a clinically and genetically heterogeneous plasma cell neoplasm. Theprognosis of MM patients is dependent on several factors including the patient’s age, the stage of disease and geneticalterations. This study aimed to determine the frequency of common chromosomal abnormalities and their significance inMM patients referred to a tertiary healthcare center in India. Methods: Fluorescence in situ hybridization on interphasenuclei from bone marrow cells using seven MM-specific probes for recurrent aberrations was performed in a total of215 newly diagnosed patients. Results: Chromosomal abnormalities were detected in 161 (74.9%) MM patients inthis study. The most frequent aberration was trisomy(ies) involving only gain of chromosomes in 48 (22.3%) cases.A translocation involving the IGH gene alone or accompanied by trisomy(ies) or by monosomy 13/13q deletion or byboth was registered in 80 (37.2%) patients. Atypical patterns such as a deletion of the IGH variable segment (IGHv)on the derivative chromosome 14 or on the native (normal) chromosome 14, biallelic deletion of IGHv, deletion ofthe IGH constant segment on the rearranged chromosome14 and extra fusions were noticed in 21 (9.8%) patientswith an IGH rearrangement. Monosomy 13/deletion 13q was identified singly or as part of a complex karyotype in74 patients (34.4%). Clonal heterogeneity and additional abnormalities including TP53 deletion and monosomies ofchromosomes 4, 9, 14 and 16 were recorded in 18.6% and 16.3% of patients respectively. Patients with abnormalitiesexhibited plasmacytosis, reduced hemoglobin value and high level of ß2-microglobulin. Conclusions: A lower medianage and a low frequency of IGH translocations particularly t(11;14) and chromosome 13 abnormalities suggest ethnicdiversity. Further investigations on genetic alterations including IGH deletions will contribute to improved insightsinto the biology of myeloma disease, risk stratification and patient management.