bcl-2与bcl-6在弥漫性大B细胞淋巴瘤中表达的研究

 目的　从蛋白水平和基因水平研究bcl-2、bcl-6在弥漫性大B细胞淋巴瘤（DLBCL）的表达。方法　应用免疫组织化学EnVision法对73例DLBCL进行CD3、CD10、CD20、bcl-2、bcl-6、MUM-1的标记。应用荧光原位杂交（FISH）技术检测其中57例t（14;18）染色体异位bcl-2基因的异常情况和54例bcl-6基因所在的3q27染色体的断裂和扩增情况。结果　肿瘤细胞表达CD10、bcl-6、MUM-1、bcl-2的阳性率分别为15.1 %、38.4 %、71.2 % 、79.2 %。57例患者中16例（28.1 %）存在t（14;18）。bcl-2蛋白表达与免疫学亚型有相关性（P＝0.035）,t（14;18）与预后有相关性（P＝0.045）,t（14;18）与bcl-2蛋白表达无相关性（P＝0.710）。54例中3q27染色体断裂11例（20.4 %）,扩增14例（25.9 %）。bcl-6阳性表达较阴性者预后好（P＝0.041）。bcl-6与3q27断裂和扩增无相关性（均P＝1.000）。结论　bcl-2、bcl-6蛋白和基因表达是各自独立的事件,它们在DLBCL中的意义不同。bcl-2蛋白是与免疫学亚型相关的预后标志物,bcl-2阳性的GCB型预后较差;t（14;18）是独立预后事件,阳性者预后较差,对靶向治疗患者应检测t（14;18）;bcl-6蛋白的表达有助于DLBCL的预后判断,可作为独立的预后因子。3q27染色体断裂可能提示预后较差。     

bcl-2与NF-κB／p65在弥漫性大B细胞淋巴瘤不同亚型中表达的意义

 目的　探讨bcl-2与NF-κB／p65蛋白在弥漫性大B细胞淋巴瘤（DLBCL）不同亚型中的表达及其意义。方法　用免疫组织化学方法检测DLBCL患者CD10、bcl-6、MUM-1蛋白的表达,以Hans等的分型原则将其划分为GCB和非GCB／ABC亚型,同时标记bcl-2与NF-κB／p65抗体,比较bcl-2与NF-κB／p65蛋白在GCB和ABC亚型中的表达情况并分析二者与DLBCL两种主要亚型生存率的相关性。结果　bcl-2与NF-κB／p65蛋白在DLBCL中的表达率为67.1 %和77.1 %, 二者表达呈正相关;GCB亚型中bcl-2与NF-κB／p65的表达率分别为52.0 %和56.0 %,ABC亚型中bcl-2与NF-κB／p65蛋白的表达率分别为75.6 %和88.9 %,ABC亚型中bcl-2与NF-κB／p65的表达率高于GCB亚型;在GCB亚型中,bcl-2与NF-κB／p65蛋白的表达与总生存率无显著的相关性,而在ABC亚型DLBCL中,bcl-2与NF-κB／p65蛋白的表达与生存率密切相关。结论　仅在ABC亚型DLBCL中,bcl-2与NF-κB／p65蛋白的表达是影响预后的重要不利因素,因此,需在DLBCL亚分型的基础上评估bcl-2与NF-κB／p65蛋白表达的预后意义。     

bcl-6、p53、c-myc基因异常在弥漫大B细胞淋巴瘤中的临床意义

目的 探讨bcl-6 、p53、c-myc基因异常的检测在弥漫大B细胞淋巴瘤（DLBCL）中的临床意义.方法 间期荧光原位杂交（I-FISH）方法检测59例DLBCL患者活体石蜡组织bcl-6、p53蛋白、c-myc基因异常的情况,同时以CHOP及R-CHOP方案化疗,评价疗效.观察bcl-6、p53蛋白、c-myc基因与化疗疗效及生存期的关系.结果 59例DLBCL中,p53丢失18例（30.5％）,bcl-6重排11例（18.6％）,c-myc重排5例（8.5％）.p53丢失阳性组化疗有效率（33.3％）明显低于阴性组（76.5％）（x2=9.560,P=0.002）. bcl-6基因重排阳性组的预后差于基因重排阴性组,但差异无统计学意义[总生存（OS）,P=0.107;无进展生存时间（PFS）,P=0.094]; p53基因丢失阳性组预后明显差于阴性组（OS,P=0.031;IPFS,P=0.028）;c-myc重排阳性组的预后差于基因重排阴性组,但差异无统计学意义（OS,P=0.163;PFS,P=0.167）.其中CHOP化疗组患者,p53基因丢失、c-myc重排阳性组的预后明显差于阴性组,差异有统计学意义（P值均＜ 0.05）;R-CHOP化疗组,bcl-6基因重排阳性组具有较差的预后意义（OS,P=0.003;PFS,P=0.007）.结论 bcl-6 、p53、c-myc基因异常与 DLBCL预后密切相关,可作为预测DLBCL的预后因素并指导治疗.     

儿童侵袭性成熟B细胞淋巴瘤的临床病理学及分子遗传学特点

 目的　分析总结中国儿童各类型侵袭性成熟B细胞淋巴瘤的临床病理学及分子遗传学特点,为其诊断的标准化提供依据。方法　收集97例儿童侵袭性成熟B细胞淋巴瘤石蜡包埋组织标本,包括伯基特淋巴瘤（BL）81例、弥漫大B 细胞淋巴瘤（DLBCL）8例、介于BL和DLBCL间的不能分类的B细胞淋巴瘤（BL／DLBCL）8例,利用免疫组织化学技术和间期荧光原位杂交（FISH）技术检测其免疫表型和分子遗传学特征。结果　BL的bcl-2和MUM1的阳性率分别为3 %（2／66）和17 %（12／71）,DLBCL分别为50 %（4／8）和63 %（5／8）,BL／DLBCL分别为 50 %（4／8）和63 %（5／8）。BL、DLBCL 和BL／DLBCL 的Ki-67平均值分别为（93±4.4）%、（83±14.3）%和（80±11.5）%。BL、DLBCL 和BL／DLBCL 的c-myc 基因易位的比例分别为98 %（79／81）、38 %（3／8）和50 %（4／8）。38 %（3／8）的DLBCL和25 %（2／8）的BL／DLBCL 存在bcl-6基因的多拷贝,BL与DLBCL 之间、BL与BL／DLBCL之间bcl-2、MUM1和 Ki-67平均值的差异及c-myc基因易位和bcl-6基因多拷贝的差异均有统计学意义（均P＜0.05）。结论　儿童侵袭性成熟B细胞淋巴瘤的诊断和分型需要综合分析形态学、免疫表型和分子遗传学特征。儿童BL／DLBCL 可能是DLBCL 的一个亚型。CD+10、bcl-6+、bcl-2-、Ki-67＞90 %、伴有IGH／c-myc重排、不伴有bcl-2和bcl-6重排时,支持BL的诊断;bcl-2+、Ki-67 为50 %～90 %,同时伴有bcl-6基因的多拷贝时,支持 DLBCL或BL／DLBCL 的诊断。     

弥漫性大B细胞淋巴瘤bcl-2、bcl-6、CD10蛋白表达及其在亚分类中的意义

目的探讨bcl-2、bcl-6、CD10在弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DL-BCL)中的表达及意义。方法应用免疫组织化学SP法,检测60例弥漫性大B细胞淋巴瘤标本中bcl-2、bcl-6、CD10蛋白的表达水平。结果bcl-2、bcl-6、CD10在60例弥漫性大B细胞淋巴瘤中的表达率分别为55.0%(33/60)、48.3%(29/60)、46.7%(28/60),其中CD10(+)/bcl-6(+)15例,25%;CD10(+)/bcl-6(-)13例,21.7%;CD10(-)/bcl-6(+)14例,23.3%;CD10(-)/bcl-6(-)18例,30%。CD10(+)/bcl-6(+)与CD10(-)/bcl-6(-)中bcl-2的表达差异具有统计学意义。结论bcl-2有望成为DLBCL亚分类的指标。     

Choi和Hans分型在弥漫大B细胞淋巴瘤预后评价中的意义

 目的　探讨Choi和Hans分型在弥漫大B细胞淋巴瘤（DLBCL）预后评价中的意义。方法　收集山西省肿瘤医院病理科有详细随访资料的DLBCL 99例,用免疫组织化学EnVision法检测bcl-2、bcl-6、CD10、FOXP1、GCET1、MUM-1的表达情况。根据Choi和Hans两种分类法分别将所有病例分型。其中35例应用荧光原位杂交技术检测bcl-6基因重排情况。结果　按Hans分类法生发中心B细胞（GCB）型21例,非GCB（nonGCB）型78例;按Choi分类法GCB型23例,nonGCB型76例。GCB型生存率明显优于nonGCB型,差异有统计学意义（P＝0.000）。FOXP1蛋白阳性表达与预后呈负相关（P＝0.011）,GCET1阳性表达则与预后呈正相关（P＝0.027）。在35例DLBCL患者中,bcl-6基因重排阳性高发于nonGCB型患者,bcl-6基因重排与bcl-6蛋白的表达没有明显相关性。结论　Choi和Hans两种分类法免疫分型GCB型预后都优于nonGCB型。bcl-6、FOXP1、GCET1的表达与预后有相关性。Choi及Hans分类法对DLBCL的免疫分型、临床预后估计均有应用价值。     

弥漫大B细胞淋巴瘤并发滤泡淋巴瘤患者临床特征、基因突变谱及预后分析

目的探讨初诊弥漫大B细胞淋巴瘤(DLBCL)并发滤泡淋巴瘤(FL)(DLBCL/FL)患者的临床病理特征、基因突变谱和预后影响因素。方法回顾性分析2018年2月至2021年11月上海交通大学医学院附属瑞金医院收治的50例DLBCL/FL患者的临床病理资料。采用靶向测序评估55个淋巴瘤相关基因的突变状态。总结患者临床特征, 评估患者近期疗效;采用Kaplan-Meier法分析患者总生存(OS)和无进展生存(PFS);采用Cox比例风险模型评估OS和PFS的影响因素。结果 50例DLBCL/FL患者中, 男性23例(46%), 国际预后指数(IPI)评分≥2分22例(44%), 16例(32%)为双表达淋巴瘤(DEL), 4例(8%)为双打击淋巴瘤(DHL)。一线治疗后患者的完全缓解(CR)率为68%(34/50), 总有效率为78%(39/50)。中位随访23.3个月(5.1～50.9个月), 2年OS率为82.1%, 2年PFS率为67.1%, 中位OS和PFS均未达到。靶向测序显示, KMT2D、MYD88、TP53、BTG2、DTX1、EZH2、CD70、CREBBP、DUSP2、...     

一种新的淋巴瘤亚型:bcl-2和myc易位的双击淋巴瘤

 双击淋巴瘤（DHL）特征介于弥漫大B细胞淋巴瘤（DLBCL）和伯基特淋巴瘤（BL）之间,通常伴有myc基因断裂和其他重现性染色体断裂的疾病,常见myc和bcl-2基因的易位。其临床表现具有乳酸脱氢酶升高、骨髓受累、Ann Abort分期晚期、B症状、结外受累、侵犯中枢神经系统等特征。因与DLBCL和BL有部分重叠,所以依靠病理诊断很难将其区分出来,目前主要的诊断方法为G显带染色体核型分析、荧光原位杂交（FISH）检测以及免疫组织化学技术。DHL对于DLBCL的标准化疗方案反应较差,预后不佳,中位生存期仅为0.2～1.5年。目前DHL尚无较好的治疗方法,主要方案为RCHOP、RICE、RCVD、甲氨蝶呤预防中枢神经系统受累、大剂量化疗联合骨髓移植等。     

弥漫大B细胞淋巴瘤患者临床病理特征及预后分析

目的探讨弥漫大B细胞淋巴瘤(DLBCL)患者的临床病理特征、分子遗传学特征及预后。方法回顾性分析北京大学第三医院及北京大学基础医学院2008年1月至2015年12月会诊及常规外检的152例DLBCL患者的临床病理资料, 采用免疫组织化学法检测CD10、bcl-6、MUM1、GCET1、FOXP1的表达情况, 采用原位杂交检测EB病毒编码小RNA;采用荧光原位杂交(FISH)检测bcl-2、bcl-6和c-myc基因异常情况, 筛选双重打击淋巴瘤(DHL)。采用Kaplan-Meier法进行生存分析。结果 152例DLBCL中, 男女比例为1.49∶1, 中位发病年龄59岁(7～90岁), 原发于淋巴结内79例(52.0%)。全部患者中位总生存(OS)时间为16个月(1～101个月), 1、3、5年OS率分别为70.2%、44.7%及30.3%。R-CHOP方案治疗组OS好于CHOP方案治疗组及未治疗组(P=0.001)。137例进行双重打击免疫组织化学评分(DHS评分)的患者中, 0分56例, 1分57例, 2分24例, 不同DHS评分组间OS差异无统计学意义(P=0.311)。FIS...     

Pokemon蛋白在弥漫性大B细胞淋巴瘤中的表达

目的:探讨Pokemon蛋白和p53、Bel-6蛋白在弥漫性大B细胞淋巴瘤(DLBCL)中的表达及相互关系.方法:采用免疫组化SP法检测Pokemon和p53、Bel-6在DLBCL中和反应性增生淋巴结(RLH)中的表达.结果:Pokemon在50例DLBCL和10例RLH中的表达分别为35例(70.0%)和1例(10.0%),Pokemon在DLBCL中的表达明显高于在RLH中的表达(P<0.01);Pokemon在20例GCB(Gernlinal Center origin ofB-cell)型(12/20,60.0%)和30例non-GCB(non-Geminal Center origin of B-cell)型DLBCL患者中(23/30,76.7%)的表达差异无统计学意义(P>0.05);而p53在GCB型DLBCL中的表达率(5/20,25.0%)明'显低于在non-CCB型DLBCL中的表达率(16/30,53.3%),P<0.05;35例Pokemon阳性DLBCL中p53阳性18例、Bel-6阳性16例,而在15例Pokemon阴性DLBCL中p53阳性3例、Bel-6阳性2例,在DLBCL中Pokemon表达分别与p53和Bcl-6的表达呈正相关.结论:Pokemon在恶性程度较高的non-GCB型的表达率较GCB型有更高趋势.Pokemon的阳性表达与突变型p53的阳性表达呈正相关,提示Pokemon蛋白和突变型p53蛋白共同参与了DLBCL的发生和发展,可能存在Pokemon-p53细胞转化通路在DLBCL的发病中起重要作用.     

Rearrangements of bcl-1, bcl-2, bcl-6, and c-myc in diffuse large B-cell lymphomas.

Diffuse large B-cell lymphoma (DLBL) is characterized by a marked degree of morphologic and clinical heterogeneity. We studied 137 patients with de novo DLBL for rearrangements of the bcl-1, bcl-2, bcl-6 and c-myc oncogenes by Southern blot analysis. Structural alterations of bcl-1, bcl-2, bcl-6, and c-myc were detected in 21 of 137 (15.3%), 8 of 137 (5.8%), 22 of 137 (16.1%), 14 of 137 (10.2%) patients, respectively. Two cases showed a combination of bcl-1 and bcl-6 rearrangements. Chromosomal analysis was performed in 31 cases of the 137 DLBL. 27 of these showed karyotypic abnormalities, and two had translocations 3q27 involving bcl-6. However, one of two cases had no rearrangement of bcl-6. Patients with rearranged bcl-6 and c-myc tended to have poorer survival than patients with germ-line. Furthermore, bcl-1 and bcl-2 rearrangements tended to have a better outcome, although the above differences were not statistically significant. Rearrangements of the bcl-1, 2, 6, and c-myc gene correlated with the clinical outcome in DLBL and may thus serve as prognostic markers in patients with this form of malignant lymphoma. However, other genetic factors are probably involved in determining prognosis.     

Patterns of Utilization and Outcomes of Autologous Stem Cell Transplantation and Chimeric Antigen Receptor T-Cell Therapy in Relapsed or Refractory Diffuse Large B-cell Lymphomas with MYC and BCL2 and/or BCL6 Rearrangements

BackgroundPatients with Diffuse Large Bcell Lymphoma (DLBCL) with MYC and BCL2 and/or BCL6 gene rearrangements [double-hit lymphoma/triple-hit lymphoma (DHL/THL)] have poor prognosis in the relapsed/refractory setting.MethodsWe utilized a real-world deidentified database of DLBCL patients and report patterns of therapy utilization in relapsed/refractory DLBCL. We used log-rank test to compare real-world overall survival (rwOS) among DHL and non-DHL subgroups for CAR Tcell therapy or ASCT respectively, stratified for prior lines of therapy.ResultsOf all 7,877 patients with DLBCL, 367 patients had DHL while 6113 had non-DHL. Second line chemotherapy was administered to 147 DHL patients and 1517 non-DHL. 1393 were excluded, including 934 with unknown DHL/THL status. Approximately 47% received salvage intent chemotherapy in the DHL subgroup, of which 19% patients eventually received ASCT, while 34% received salvage intent chemotherapy in the non-DHL/THL group with 32% receiving ASCT. DHL/THL status negatively influenced median rwOS for patients who underwent ASCT in the second-line while it was associated with numerically inferior but without statistically significant rwOS among patients that underwent CAR Tcell therapy on multivariable analysis.ConclusionrwOS of relapsed DHL/THL is inferior to non-DHL/THL. Fewer patients with DHL/THL were able to proceed with ASCT after salvage chemotherapy compared to non-DHL/THL. ASCT as second-line therapy for relapsed DHL/THL had worse rwOS than for non-DHL/THL, consistent with the natural history of DHL/THL. This difference was not seen for CAR Tcell therapy, which combined with promising results from clinical trials, suggests a greater role for CAR T-cell therapy in relapsed/refractory DHL.     

双重打击淋巴瘤研究进展

双重打击淋巴瘤(DHL)通常是指发生MYC基因重排,同时伴有bcl-2和(或)bcl-6基因重排的大B细胞淋巴瘤.DHL侵袭性强,常规方案治疗预后差,目前尚无标准的一线治疗方案.文章就DHL在诊断、生物学行为、预后影响因素和治疗方面的研究进展进行综述,旨在加深对DHL的理解,优化治疗策略,以期改善其预后.     

High levels of bcl-2 protein expression do not correlate with genetic abnormalities but predict worse prognosis in patients with lymphoblastic lymphoma

We aimed to investigate bcl-2, bcl-6, and c-myc rearrangements in patients with lymphoblastic lymphoma (LBL), especially focus on the correlation of protein expression with genetic abnormalities. Moreover, their prognostic significance was further analyzed in LBL. Protein expression and genetic abnormalities of bcl-2, bcl-6, and c-myc were investigated in microarrayed tumors from 33 cases of T cell LBL and eight cases of B cell lineage. Immunohistochemical (IHC) staining was performed to evaluate protein expression, including bcl-2, bcl-6, c-myc, TdT, CD1α, CD34, Ki-67, PAX-5, CD2, CD3, CD4, CD8, and CD20. Genetic abnormalities of bcl-2, bcl-6, and c-myc were detected by dual color fluorescence in situ hybridization (FISH). Bcl-2 protein was positive in 51.2 % (21/41) of the patients, bcl-6 protein in 7.3 % (three out of 41), and c-myc protein in 78.0 % (32/41). Bcl-2 breakpoint was found in two cases by FISH analysis. There was no evidence of bcl-6 or c-myc rearrangement in patients with LBL. However, both gene gain and loss events occurred in bcl-2, bcl-6, and c-myc. A univariate analysis showed that stage III or IV, elevated lactate dehydrogenase (LDH), and positivity for bcl-2 protein were associated with shorter survival (p?<?0.05). Enhanced protein expression and detectable genetic abnormalities of bcl-2, bcl-6, and c-myc were observed in patients with LBL. No statistical correlation was found between IHC results and cytogenetic findings. Stage III or IV, elevated LDH, and positivity for bcl-2 protein were identified as adverse prognostic factors. The patients with more adverse factors would have increasingly worse prognosis.     

Leukemic High Grade B Cell Lymphoma is Associated With MYC Translocation,Double Hit/Triple Hit Status,Transformation, and CNS Disease Risk: The Mayo Clinic Experience

Introduction: Leukemic involvement in high grade B cell lymphoma (L-HGBL) is rare and has been sparsely described in the literature. We report our experience in a large single institution multicenter academic setting. Materials and Methods: Medical records of patients with HGBL who received care at Mayo Clinic between 2003 and 2020 were reviewed. L-HGBL was confirmed by peripheral blood smear and flow cytometry with corroboration from tissue and bone marrow biopsy findings. Results: Twenty patients met inclusion criteria. All patients had significant bone marrow involvement by HGBL. Leukemic involvement presented in 11 of 20 (55%) in the de novo and 9 of 20 (45%) in the relapsed setting. Seven of 20 patients had DLBCL, NOS, 6 of 20 had transformation (t-DLBCL), 3 of 20 had transformed double/triple hit lymphoma (t-DHL/THL), 2 of 20 had double hit lymphoma (DHL), and 2 of 20 had HGBL with intermediate features between DLBCL and Burkitt lymphoma. Nine of 15 patients had MYC translocation. Based on Hans criteria, 11 of 20 had germinal center B-cell (GCB) cell of origin (COO) and 9/20 had non-GCB COO. Five of 11 de novo patients experienced CNS relapse/progression. All de novo patients received anthracycline-based chemoimmunotherapy. Eighteen of 20 patients died of progressive disease. Median overall survival was significantly better in the de novo compared to relapsed group (8.9 months vs. 2.8 months, P = .01). COO, MYC status, DHL/THL status, HGBL subtype, or treatment group did not demonstrate a significant effect on overall survival. Conclusion: L-HGBL carries a poor prognosis and is associated with MYC translocation, DHL/THL status, transformation, and high CNS risk. Novel therapeutic approaches are needed for L-HGBL.     

219例弥漫性大B细胞淋巴瘤免疫表型及遗传学特征分析

  目的  探讨国内弥漫性大B细胞淋巴瘤（DLBCL）免疫表型分型及BCL-2和BCL-6基因异常的分布情况。  方法  应用组织芯片和免疫组织化学法及FISH技术对219例DLBCL的免疫表型及BCL-2和BCL-6基因异常进行检测,根据Hans法进行分型;并收集国内7家相关研究报道,进行综合分析。  结果  本组研究结果:219例DLBCL中,非GCB型（165例,75.3%）显著高于GCB型（54例,24.7%）（P < 0.001）。BCL-2基因异常共49例（25.8%）,其中t （14;18）5例（2.6%）均为GCB型;BCL-2基因扩增44例（23.2%）,GCB型4例（8.5%）,非GCB型40例（28.0%）,有显著性差异（P=0.013）。BCL-6基因重排共42例（22.1%）,GCB型7例（14.9%）,非GCB型35例（24.5%）,差异无统计学意义（P=0.189）。BCL-2基因扩增和BCL-6基因重排呈显著负相关（r=-0.180,P=0.013）。8组综合分析:1 259例中非GCB型（879例,69.8%）明显高于GCB型（380例,30.2%）（P < 0.001）;免疫表型中CD10和MUM1阳性率组间差异较小（P=0.047和P=0.048）,而BCL-6及BCL-2阳性率组间存在明显差异（P < 0.001）。  结论  我国DLBCL患者在主要免疫表型和遗传学特征方面具有独特性,对此值得进行深入研究。