荧光原位杂交技术检测多发性骨髓瘤染色体

目的研究荧光原位杂交技术(FISH)在多发性骨髓瘤(MM)染色体异常检测中的应用价值。方法选取从2012年1月至2014年1月该院收治的初治MM患者24例,同时采用常规G显带技术与FISH技术进行检测,比较两种检测方法的阳性检出率,分析染色体异常与MM的免疫学分型、临床分期与预后的相关性。结果常规G显带技术的染色体异常检出率比FISH检出率低(P0.01);常规G显带技术仅检出1例染色体异常者,MM患者的染色体异常以Ⅲ期和Ig G型为主。结论 FISH技术的染色体异常检出率与特异性较高,临床分期、分型与染色体异常存在相关性,Ⅲ期和Ig G型患者出现染色体异常的可能性更高。     

多发性骨髓瘤患者81例细胞及分子遗传学研究

目的 探讨多发性骨髓瘤(MM)患者遗传学特征,评价添加细胞因子及延长培养时间对MM染色体核型分析的影响和间期荧光原位杂交(FISH)检测RB1与P53基因缺失的意义.方法 采用骨髓24h短期培养和G显带技术对81例MM患者进行染色体核型分析,其中28例患者同时采用添加IL-6(终浓度10 μg/L)和重组粒-巨噬细胞集落刺激因子(GM-CSF,终浓度40 μg/L)的6d培养法后的G显带核型分析,31例患者采用间期FISH方法检测RB1与P53基因缺失.结果 81例患者中有75例患者有足够可供分析的中期分裂象,其中31例(41.3％)检出异常克隆.在31例检出染色体异常核型的患者中,单纯染色体数目异常4例(12.9％),单纯染色体结构异常11例(35.5％),染色体数目和结构异常同时存在16例(51.6％).28例患者同时采用两种培养方法对照研究结果显示,骨髓24h短期培养法异常克隆检出率为25.0％,添加细胞因子的6d延期培养法异常克隆检出率为51.9％ (P =0.026).31例患者间期FISH结果显示RB1基因缺失10例(32.3％),P53基因缺失11例(35.5％),5例患者RBI和P53基因均缺失.结论 MM染色体核型异常半数以上为染色体数目和结构异常同时存在,显带技术是遗传学检测的基础,添加细胞因子的长期培养法可提高显带技术的异常克隆检出率,间期FISH是一种检测MM患者骨髓瘤细胞基因缺失的敏感方法,具有临床应用价值.     

21例多发性骨髓瘤分子遗传学异常的FISH检测的意义

目的：为探讨间期荧光原位杂交（FISH）在检测多发性骨髓瘤（MM）间期细胞13q14缺失、1q21、p53缺失以及免疫球蛋白重链（IgH）基因重排中的意义。方法：采用组合探针（1q21/RB1、D13S319/p53、IgH）对21例MM患者骨髓进行FISH检测,分析其分子遗传学异常,比较其与常规染色体检查及临床指标的相关性。结果：21例MM患者中,19例（90.48%）检测出1种或1种以上的细胞遗传学异常,15例（71.43%）同时检测出2种及以上的异常。其异常比例从高到低分别为：＋1异常（66.67%）,IgH基因重排（57.14%）,13号染色体缺失（47.62%）和p53基因丢失（23.81%）。3例（14.29%）通过G-显带常规染色体检查发现异常,与FISH比较两者差异有统计学意义（P〈0.01）。结论：＋1、IgH基因重排及13q14缺失在MM中的发生率较高。FISH技术能提高MM分子遗传学异常的敏感性。     

多发性骨髓瘤浆细胞标记指数检测的临床意义研究

目的探讨多发性骨髓瘤（MM）中浆细胞标记指数（PCLI）检测的临床意义及与年龄、疾病分期、骨髓浆细胞数、β2-微球蛋白（β2-MG）、白蛋白、乳酸脱氢酶（LDH）、血肌酐、生存期及13q14缺失[del（13q14）]等的相关性。方法应用免疫荧光玻片计数法检测42例初诊MM患者PCLI,并应用间期荧光原位杂交（I-FISH）技术,用位于13q14的序列特异性DNA探针D13S319,检测患者del（13q14）异常情况。结果 42例初诊MM患者中,PCLI阳性18例（42.9%）,其中del（13q14）阳性14例,阴性4例;PCLI阴性24例（57.1%）,其中del（13q14）阳性12例,阴性12例,PCLI高表达患者具有更高的del（13q14）发生率（P〈0.01）。PCLI阳性与年龄、疾病分期、骨髓浆细胞数、β2-MG、白蛋白、LDH、肌酐等无相关性（P值均〉0.05）,与疾病无进展生存期（PFS）短有关（P〈0.05）。结论 PCLI是判断MM患者预后的重要指标,阳性患者,疾病进展快,生存期短。PCLI阳性者del（13q14）发生率高。     

92例多发性骨髓瘤的临床与细胞遗传学分析

目的探讨多发性骨髓瘤（MM）细胞染色体异常的临床意义。方法采用骨髓细胞直接法、24h短期培养法制备染色体标本,用R显带技术进行核型分析。结果92例MM中,常规细胞遗传学（CC）发现异常核型24例,发生率26．09％。部分病例在CC基础上应用荧光原位杂交（FISH）技术,异常检出率大为提高。结论FISH技术深入了MM的细胞遗传学研究,伴有14q32异常是MM患者最为常见的染色体核型异常,伴有多条染色体异常MM患者其临床预后差。     

16例多发性骨髓瘤患者的染色体分析

目的 :分析多发性骨髓瘤 (MM )细胞遗传学变化及其与MM的分型、分期及预后的关系。方法 :采用短期培养法及G显带技术对 16例MM患者进行细胞染色体核型分析 ,且对MM分型、分期及预后进行相关性分析。结果 :在 16例MM患者中发现染色体异常者 10例 (占 6 2 .5 % ) ,均有数目异常 ,表现有超二倍体、亚二倍体、多倍体及假二倍体 ;其中 9例发生于MMⅢ期 (占 5 6 .2 5 % ) ,1例发生于MMⅠ期 (占 6 .2 5 % ) ;4例为IgG型 ,3例轻链型 ,2例为IgA型 ,1例为不分泌型 ;有异常核型的MM患者表现为高 β2 微球蛋白 (β2 M )、高C反应蛋白(CRP)、低血红蛋白 (Hb)、低血小板 (PLT)、骨髓瘤细胞计数超过 30 % ;高Ca2 + 、高乳酸脱氢酶 (LDH) ,与无核型改变的MM患者比较差异有统计学意义 (P <0 .0 5 )。结论 :大多数MM患者存在染色体异常 ,且与临床分期及预后有关系 ,多发生于Ⅲ期 ,预后差 ,与其免疫分型无关     

多发性骨髓瘤细胞遗传学变化及其蛋白酶体抑制剂的疗效观察

目的分析多发性骨髓瘤(MM)的细胞遗传学变化及其与蛋白酶体抑制剂治疗的关系。方法对2005年1月至2006年7月北京大学人民医院29例初诊的MM患者行染色体核型检查,采用骨髓细胞24h短期培养,用常规方法制备染色体,G显带进行核型分析。22例MM患者采用传统化疗即长春新碱联合阿霉素和地塞米松(VAD)方案或马法兰与泼尼松(MP)方案;7例患者应用蛋白酶体抑制剂(硼替佐米)为主的化疗方案。比较两组患者的有效率及缓解率。结果29例MM患者中异常核型检出率为37.9%,其中有复杂和高度复杂畸变的占81.8%。采用VAD或MP方案化疗的核型正常组的有效率为81.2%,核型异常组有效率为0,差异有显著性意义(P<0.05)。应用硼替佐米为主的化疗方案中核型异常组5例均有效,核型正常组2例均有效。核型异常组硼替佐米有效率与传统化疗组相比,差异有显著性意义(P<0.05)。结论染色体核型异常的患者,应首选蛋白酶体抑制剂硼替佐米等进行治疗。     

细胞肿瘤高危遗传学异常在多发性骨髓瘤危险度分层中的应用

目的在对多发性骨髓瘤危险度进行分层的过程中进行细胞肿瘤高危遗传学异常分析,以更好地进行临床分层。方法利用荧光原位杂交法对初治多发性骨髓瘤及人多发性骨髓瘤细胞系的分子遗传学异常进行检测和比较。结果细胞遗传学总体检出率为85.54%(71/83),在分子遗传学异常方面,大部分初治多发性骨髓瘤、全部人多发性骨髓瘤细胞系均存在,但分子遗传学异常无统计学差异(P0.05)。1例为人多发性骨髓瘤细胞系伴有IGH/CCND1,且IGH/FGFR3阳性检出率显著高于初治多发性骨髓瘤(P0.05)。6例人多发性骨髓瘤细胞系具有1q21的扩增,但与初治多发性骨髓瘤无统计学差异(P0.05)。在RB-1缺失方面,人多发性骨髓瘤细胞系与初治多发性骨髓瘤差异不显著(P0.05);P53缺失存在统计学差异(P0.05),IGH/MAF阳性有统计学差异(P0.05);在1q21扩增和IGH异常、IGH/CCND1阳性差异均不显著(P0.05)。结论浆细胞肿瘤恶性程度的最高形式是人多发性骨髓瘤细胞系,人多发性骨髓瘤细胞系积累了大量的遗传学异常,可为多发性骨髓瘤危险度分层提供一定的依据。     

多发性骨髓瘤发病的分子生物学机制

多发性骨髓瘤(MM)是浆细胞恶性肿瘤,其瘤细胞是终末分化的B淋巴细胞。最近,在MM中发现大量的染色体异常,最明显的是13q缺失和免疫球蛋白重链基因易位。易位的伙伴染色体常是癌基因,易位后激活癌基因。另外,抑癌基因如N-ras失活在MM发生、发展中也起重要作用。本文就此讨论MM中常见的几种染色体异常。     

多发性骨髓瘤预后分层研究进展

多发性骨髓瘤(multiple myeloma,MM) 是一种浆细胞恶性增殖性疾病,生物学异质性强,个体间临床疗效差异 大,生存期从数月至数十年不等。构建准确的预后分层评估体系已成为MM 临床研究的发展趋势。分子遗传学是 影响MM 预后的主要内在因素,建立以分子遗传学为基础的新预后分层模式,综合疾病的宿主因素、肿瘤负荷、肿瘤 生物学因素等各项指标及治疗反应的预后分层从而进行个体化治疗对现代MM 治疗非常必要。     

Recent advances in cytogenetic characterization of multiple myeloma

The detection of cytogenetic abnormalities in multiple myeloma (MM) has received more importance over last years for risk stratification and the new risk‐adapted treatment strategies. Conventional G‐banding analysis should be included in a routine procedure for the initial diagnostic workup for patients suspected of MM. However, the detection of chromosomal abnormalities in MM by conventional cytogenetics is limited owing to the low proliferative activity of malignant plasma cells as well as the low number of plasma cells in bone marrow specimens. Fluorescence in situ hybridization (FISH) or microarray‐based technologies can overcome some of those drawbacks and detect specific target arrangements as well as chromosomal copy number changes. In this review, we will discuss different cytogenetic approaches and compare their strength and weakness to provide genetic information for risk stratification and prediction of outcome in MM patients.     

Cytogenetics of multiple myeloma: interpretation of fluorescence in situ hybridization results

The cytogenetic picture in multiple myeloma (MM) is highly complex, from which non-random numerical and structural chromosomal changes have been identified. Specifically, translocations involving the immunoglobulin heavy chain gene (IGH) at 14q32 and either monosomy or deletions of chromosome 13 have been reported in a significant number of patients from both cytogenetic and interphase fluorescence in situ hybridization (FISH) studies. Importantly, these abnormalities of chromosome 13 have recently been associated with a poor prognosis. In view of the highly complex nature of the karyotypes in MM patients, interphase FISH results may be difficult to interpret. In this study, cytogenetics and/or interphase FISH were carried out on bone marrow samples or purified plasma cells from 37 MM patients. Abnormal karyotypes, characterized by multiplex FISH (M-FISH) were found in 11 patients, all of which were highly complex. Interphase FISH revealed translocations involving the IGH locus in 16 (43%) patients. The IGH/cyclin D1 (CCND1) gene fusion characteristic of the translocation, t(11;14)(q13;q32), was seen in 12 (32%) of these patients and other rearrangements of IGH in four (11%) patients. Fourteen patients had additional copies of chromosome 11. Twenty patients (54%) had 13q14 deletions, 10 of whom also had t(11;14) or another IGH translocation. By comparing cytogenetic and FISH results, this study has revealed that significant chromosomal abnormalities might be hidden within highly complex karyotypes. Therefore, extreme caution is required in the interpretation of interphase FISH results in MM, particularly in relation to certain abnormalities, such as 13q14 deletions, which have an impact on prognosis.     

P53 deletion is not a frequent event in multiple myeloma

Recently a high incidence of interstitial deletion of the P53 locus has been reported in multiple myeloma (MM) patients. Considering the importance of such an event, we analysed 79 patients with advanced-stage disease using fluorescence in situ hybridization (FISH). Strikingly, we found only 7/79 patients with a P53 deletion. In order to rule out any differences in probe selection, we reanalysed all the patients with the same probe as that used in a previous study, and confirmed the low incidence of P53 deletion (7/79, 9%). The only explanation is a difference in hybridization efficiency. Since hybridization is far less efficient on malignant plasma cells than on other bone marrow cells we suggest that this poor hybridization efficiency may lead to a false P53 deletion.     

Management of multiple myeloma

Summary There has been little progress in the treatment of patients with multiple myeloma, and the average survival time is still only about 3 years. Although there have been significant therapeutic advances in recent years, clinical trials have only just begun. The major concern is, of course, the achievement of major disease control (which can be equated with a cure). The data available to date indicate that this is possible only with the use of allogeneic bone marrow transplantation, with which a survival plateau of around 30% can be attained. The trials should perhaps include the sequential use of all regimens with established efficacy in refractory myeloma. Immunoconjugate therapy with either radioisotopes or cytotoxic agents could also be envisioned, and expansion with suitable biological agents such as interleukin-2 could be considered. There is a plethora of promising treatment possibilities and novel concepts that may improve the dismal outlook for patients with multiple myeloma.Supported in part by grants CA28771 and CA37161 from the National Cancer Institute, National Institutes of Health, Bethesda, MD 20205, USA     

c-myc overexpression is not mandatory in aggressive-phase multiple myeloma with Burkitt's type translocation

Received: 6 September 1999 / Accepted: 23 December 1999     

多发性骨髓瘤92例临床分析

目的　对多发性骨髓瘤(MM)的临床特点进行总结,为诊断和治疗提供参考。方法　对92例MM患者的临床资料进行回顾分析。结果　(1)初始化疗采用M2、VAD或MP方案有效率无显著性差异。(2)初始VAD方案化疗是否有效与患者预后相关。初治有效组和无效组中位生存期分别为43. 1个月与4. 0个月(P<0. 05)。(3)骨膦可防治骨骼进一步的破坏,但骨X线片未显示明显的骨质再钙化。化疗中加用骨膦治疗可延长患者的生存期(P<0. 05)。结论　初始VAD方案化疗有效者预后较好,中位生存期可达43. 1个月,加用骨膦治疗可延长生存期。在细胞遗传学和基因表达谱基础上行个体化治疗可能会提高疗效。     

Hyperphosphatemia in multiple myeloma

Summary We report three cases of IgG kappa multiple myeloma with pseudohyperphosphatemia. The patients' serum calcium levels were normal, and the hyperphosphatemia was not related to impaired renal function. No hypoparathyroidism was found, and no exogenous phosphate preparation had been given. Since the hyperphosphatemia was of no obvious clinical or physiological significance, as evidenced by normal serum levels of 1,25 dihydroxy vitamin D₃, it was diagnosed as spurious and was connected to interference of the paraprotein with the chromogenic assay. In two of the patients major fluctuations in serum phosphate levels were seen, induced by the changes in globulin and paraprotein levels that occurred during therapy and relapse.