Ibrutinib抑制弥漫大B细胞淋巴瘤细胞生存的作用研究

  目的  探讨Bruton酪氨酸激酶（Bruton's tyrosine kinase,BTK）抑制剂ibrutinib抑制弥漫大B细胞淋巴瘤（diffuse large B cell lymphoma,DLBCL）细胞生存的作用及其相关机制。  方法  用不同浓度的ibrutinib处理DLBCL细胞系SUDHL-10、HBL-1和患者原代细胞,以MTT法检测细胞的增殖抑制情况;以Annexin V/PI流式细胞术和DAPI染色法检测细胞的凋亡情况;应用Western blot法检测细胞表达磷酸化BTK、AKT、ERK的变化。DLBCL细胞与MSC共培养后,体外克隆形成实验和NOD/SCID肿瘤模型小鼠检测ibrutinib在肿瘤微环境里对DLBCL细胞的抑制作用。  结果  2.5μmol/L及更高浓度的ibrutinib对DLBCL细胞的增殖有明显的抑制作用,且呈剂量依赖性。1.0、2.5μmol/L ibrutinib作用于SUDHL-10细胞24 h,细胞凋亡率分别为（21.73±3.64）%和（34.71±2.36）%,高于对照组（3.55±1.89）%（P < 0.05）。5、10μmol/L ibrutinib处理24 h后,DLBCL细胞系均出现核皱缩（5μmol/L）、碎裂（10μmol/L）。ibrutinib处理细胞后磷酸化BTK、AKT、ERK的表达均明显降低。ibrutinib抑制共培养时DLBCL细胞的体外克隆形成（P < 0.01）及DLBCL细胞在体内的增殖生长,差异均具有统计学意义（P < 0.05）。  结论  ibrutinib可抑制细胞系SUDHL-10和HBL-1的增殖,诱导凋亡,其机制可能通过阻断AKT、ERK信号途径而实现;在肿瘤微环境中ibrutinib同样对DLBCL细胞具有较强的抑制生存的作用,该药物有望为DLBCL的治疗带来希望。      

Smac mimetics and TRAIL cooperate to induce MLKL-dependent necroptosis in Burkitt's lymphoma cell lines

Burkitt''s lymphoma (BL) is a highly aggressive form of B-cell non-Hodgkin''s lymphoma. The clinical outcome in children with BL has improved over the last years but the prognosis for adults is still poor, highlighting the need for novel treatment strategies. Here, we report that the combinational treatment with the Smac mimetic BV6 and TRAIL triggers necroptosis in BL when caspases are blocked by zVAD.fmk (TBZ treatment). The sensitivity of BL cells to TBZ correlates with MLKL expression. We demonstrate that necroptotic signaling critically depends on MLKL, since siRNA-induced knockdown and CRISPR/Cas9-mediated knockout of MLKL profoundly protect BL cells from TBZ-induced necroptosis. Conversely, MLKL overexpression in cell lines expressing low levels of MLKL leads to necroptosis induction, which can be rescued by pharmacological inhibitors, highlighting the important role of MLKL for necroptosis execution. Importantly, the methylation status analysis of the MLKL promoter reveals a correlation between methylation and MLKL expression. Thus, MLKL is epigenetically regulated in BL and might serve as a prognostic marker for treatment success of necroptosis-based therapies. These findings have crucial implications for the development of new treatment options for BL.     

甲基转移酶抑制剂抑制弥漫大B细胞淋巴瘤细胞增殖及机制研究

目的:探索甲基转移酶抑制剂DCG066对人弥漫大B细胞淋巴瘤细胞TMD-8的有效杀伤浓度,进而对其作用机制进行研究.方法:通过CCK-8实验探索DCG066对TMD-8细胞的有效杀伤浓度,通过免疫荧光实验分析DCG066处理后TMD-8活细胞及死细胞的分布,通过流式细胞术检测DCG066对TMD-8细胞凋亡的诱导、细胞...     

弥漫性大B细胞淋巴瘤诊治进展

弥漫性大B细胞淋巴瘤(DLBCL)是目前最常见的成人非霍奇金淋巴瘤,不同亚型在临床表现、遗传学以及分子生物学等特征方面存在明显差异,所观察到的遗传学改变主要集中在BCL6,BCL2,cMYC,FAS(CD95)的突变和体细胞超突变的异常上。虽然CHOP方案化疗能够治愈部分DLBCL,但完全缓解率仅为45%~55%。强化化疗方案或干细胞移植可改善某些患者的长期生存,最突出和一致改善弥漫性大B细胞淋巴瘤的长期生存效果则是随着抗CD20单克隆抗体药物与化疗的联合应用。     

ALK+弥漫大B细胞淋巴瘤的研究进展

ALK+弥漫大B细胞淋巴瘤(anaplastic lymphoma kinase-positive diffuse large B-cell lymphoma, ALK+DLBCL)是弥漫大B细胞淋巴瘤的一种罕见的亚型, 具有特殊的免疫母细胞或浆母细胞样的形态学特点, 免疫表型独具特征, 细胞遗传学异常, 在儿童和成人都可发生。此病虽然罕见, 但是其临床过程具有侵袭性且预后不良, 因此明确认识该疾病的特点是诊断的关键。ALK+DLBCL对传统化疗方案反应性差, 最近推出的小分子ALK抑制剂可能对这种疾病的患者提供了一个潜在的新的治疗选择。      

Reversible posterior leukoencephalopathy syndrome following CHOP chemotherapy for diffuse large B-cell lymphoma

A 63-year-old female with stage IE diffuse large B-cell lymphomadeveloped reversible posterior leukoencephalopathy syndrome (RPLS)following CHOP chemotherapy, with typical clinical and radiologicalfindings. RPLS is a rare neurological syndrome characterised by visualdisturbances, seizures, headaches and altered conscious level which hasbeen associated with malignant hypertension, pre-eclampsia and somedrugs, including ciclosporin. It has not been previously reportedfollowing CHOP chemotherapy. Alternative treatment should be consideredfor patients who develop this rare complication.     

Rituximab and chemotherapy in diffuse large B-cell lymphoma

Rituximab is an anti-CD20 chimeric monoclonal antibody with activity in nearly all subtypes of B-cell lymphomas. Association of rituximab with chemotherapy (mostly the cyclophosphamide, doxorubicin, vincristine and prednisolone [CHOP] regimen) in diffuse large B-cell lymphoma (DLBCL) represents an extraordinary revolution in the prognosis of DLBCL, and is the new standard of therapy in elderly and young, low-risk patients. Despite the lack of randomized, clinical trials in younger patients with high risk, rituximab is also a standard of care in these patients in clinical practice, at least in North America. The practice is based on observational trials (e.g., the British Columbia Registry) and the missing logic in classifying patients as ‘younger’ or ‘older’: 60 years old or 65 years old. In Europe, trials are ongoing to establish the best treatment for young, high-risk patients. Association of rituximab and chemotherapy deeply modifies prognostic factors defined before the rituximab era.     

白蛋白与纤维蛋白原的比值预测弥漫大B细胞淋巴瘤患者预后的价值

目的:探讨血清白蛋白（albumin,ALB）与血浆纤维蛋白原（fibrinogen,FIB）的比值（ALB to FIB ratio,AFR）对弥漫大B细胞淋巴瘤（diffuse large B-cell lymphoma,DLBCL)患者预后的影响。方法:选择我院2015-04-17至2022-03-18确诊的DLBCL病例59例,收集所有患者首次化疗前1周内的血清ALB值、血浆FIB值,计算出AFR。应用生存分析研究AFR对DLBCL患者无进展生存率(progression free survival,PFS)的影响。应用受试者工作特征曲线(receiver operating characteristic curve,ROC)下的面积（area under the ROC curve,AUC）来评估各模型对DLBCL患者预后预测能力的大小。结果:AFR范围3.87～20.13,中位数11.16。AFR与IPI、Ann Arbor分期、结外侵犯数以及B症状密切相关。Kaplan-Meier生存分析提示,AFR增高时,DLBCL患者的PFS显著提高（P＜0.001）,2年累积PFS与低AFR组患者相比提高了73.0%。Cox单因素分析提示,低AFR的DLBCL患者发生肿瘤进展或死亡的风险显著增加（P=0.002）;多因素分析提示,AFR是影响DLBCL患者PFS的独立因素（P=0.004）。国际预后指数（international prognostic index,IPI）和IPI联合AFR两种模型判断DLBCL患者PFS的AUC值分别为0.778（95%CI 0.659～0.897,P＜0.001）和0.829（95%CI 0.723～0.935,P＜0.001）。结论:AFR是判断DLBCL患者PFS的独立预测因子,IPI联合AFR能够更好地判断DLBCL患者的预后。     

弥漫性大B细胞淋巴瘤免疫治疗进展

弥漫性大B细胞淋巴瘤（diffuse large B-cell lymphoma,DLBCL）是最常见的非霍奇金淋巴瘤（NHL）亚型,其具有高度异质性和侵袭性。尽管许多患者应用R-CHOP(利妥昔单抗+环磷酰胺+阿霉素+长春新碱+泼尼松)方案一线治疗后达到完全缓解（CR）,但仍有部分患者之后发展为复发和难治性的DLBCL,而一旦发展为复发难治性的DLBCL,常规的放疗和化疗则收效甚微。近年来,免疫治疗逐渐成为研究热点,如单克隆抗体治疗、双特异性抗体治疗、抗体-药物偶连物（ADC）治疗和嵌合体抗原受体修饰T细胞（CAR-T）治疗等。本文现就弥漫性大B细胞淋巴瘤免疫治疗进展进行综述。     

利妥昔单抗联合CHOP方案对弥漫型大B细胞淋巴瘤的疗效及影响因素

目的探讨利妥昔单抗联合CHOP方案对弥漫型大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)的疗效,观察影响化疗疗效的相关因素。方法本组选择我院2006年1月至2011年1月收入的DLBCL患者17例,患者给予利妥昔单抗联合CHOP方案治疗。观察化疗后疗效及淋巴结亚群改变情况,收集患者的性别、年龄、PS评分、Ann Arbor分期、结外浸润情况、LDH水平、体力状态(PS)评分、B症状、血红蛋白及T细胞浸润等相关情况,观察其对化疗的相关影响。结果本组所有17例患者完成治疗,治疗后疗效评估显示CR7例,PR6例,SD3例,PD1例,治疗有效率为76.4%(13/17)。影响化疗疗效相关因素分析中显示,血清LDH水平、PS评分、AnnArbor分期为影响患者化疗疗效的相关因素,P0.05。结论利妥昔单抗联合CHOP对弥漫型大B细胞淋巴瘤疗效显著,其中PS评分低、Ann Arbor分期低、血清LDH水平正常的患者化疗疗效相对较好。     

微小RNA与弥漫大B细胞淋巴瘤

微小RNA（miRNA）是一类小分子非编码RNA,通过与目标mRNA互补序列结合,在转录水平调控基因的表达。研究发现,包括弥漫大B细胞淋巴瘤在内的不同恶性肿瘤中存在特定的miRNA表达谱。因此,体液、组织标本中miRNA的表达将有望成为评估弥漫大B细胞淋巴瘤的新指标。     

弥漫性大B细胞淋巴瘤基因分型研究进展

弥漫性大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)在临床特征、基因表达和治疗反应及预后方面均表现出明显异质性.目前,R-CHOP(利妥昔单抗、环磷酰胺、阿霉素、长春新碱、强的松)作为标准治疗方案已不能满足临床需要,而近年的几项基于细胞起源分型进行的R-CHOP+X临床试验均未...     

Long-term spontaneous regression of Stage IV diffuse large B-cell lymphoma

Diffuse large B cell lymphoma (DLBCL) is an aggressive disorder accounting for >30% of all lymphomas. Its prognosis is poor due to a high relapse rate. Spontaneous regression (SR) in DLBCL is rare, with only a few reported cases. Moreover, almost all of these were low-grade lymphomas with an average SR duration of 13 mo. As the cause of SR is unknown, there are many theories such as trauma, infection, medication, and an antitumor immune response. We present a patient with progressive DLBCL who demonstrated SR for >42 mo. Although treatment for lymphoma usually starts soon after diagnosis, insights into SR of lymphomas may lead to new treatment strategies.     

EB病毒阳性的胃弥漫大B 细胞淋巴瘤特点及预后

目的:探讨EB病毒阳性患者的胃弥漫大B 细胞淋巴瘤（diffuse large B-celllymphoma,DLBCL ）病理学特点及预后。方法:回顾性分析北京大学基础医学院病理学系2009年1 月至2015年1 月75例胃弥漫大B 细胞淋巴瘤患者的临床资料,15例EB病毒（Epstein-Barr virus,EBV ）阳性者为病例组,60例EBV 阴性者为对照组,采用免疫组织化学法和EB病毒RNA 探针原位杂交法检测Bcl- 2、c-myc 蛋白表达及EBV-EBER 情况,分析EBV 阳性的胃DLBCL 患者的病理学特点及预后。结果:EBV 阳性组在临床表现、年龄、性别、起源、细胞形态等方面与EBV 阴性组相比,差异无统计学意义（P > 0.05）;在Bcl- 2、c-myc 蛋白表达方面,EBV 阳性组与EBV 阴性组相比,差异无统计学意义（P > 0.05）;R-CHOP 方案治疗下,EBV 阳性组与EBV 阴性组相比,中位总生存期（median overallsurvival,OS）分别为15.1 个月和31.4 个月,差异具有统计学意义（P = 0.01）。 结论:发生于胃DLBCL 患者中,EB病毒感染对临床表现、瘤细胞的起源、形态、蛋白表达等方面无明显影响;EB病毒阳性的DLBCL 患者并不局限于老年人;R-CHOP 治疗下EB病毒阳性患者的预后比EB病毒阴性的患者预后差。      

Expression of PLK1 and survivin in diffuse large B-cell lymphoma

Polo-like kinase 1 (PLK1) belongs to a conservative family of serine/threonine kinase and plays an important role in the process of cell cycle. Survivin is a member of inhibitor of apoptosis protein (IAP) family. We investigated the expression of PLK1 and survivin with immunohistochemical techniques in diffuse large B-cell lymphoma (DLBCL) and assessed their significance as a potent new tumor marker. The expression rate of PLK1 and survivin were 66.7% (26/39) and 82.1% (32/39), respectively. PLK1 expression correlated with systemic symptom, LDH level, IPI scores and therapeutic effect in DLBCL, while survivin did not. PLK1 expression correlated with shortened event-free survival (EFS) using the Log-rank test in DLBCL, but survivin did not. Cox regression analysis identified the independent prognostic significance for PLK1. The results suggest that there is a significant relationship between over expression of PLK1, the clinical features and survival time. Compared with survivin, PLK1 seems to be a better independent prognostic factor for DLBCL.     

沉默GCB型弥漫大B细胞淋巴瘤细胞中AFAP1-AS1的表达对细胞增殖和凋亡的影响

目的:探讨沉默GCB弥漫型大B细胞淋巴瘤（diffuse large B cell lymphoma,DLBCL）中AFAP1-AS1的表达对细胞增殖和凋亡的影响。方法:培养GCB-DLBCL细胞至对数生长期后转染OCI-Ly1细胞系,建立的GCB-DLBCL细胞系对AFAP1-AS1表达进行沉默;实验设立3组,实验组为腺病毒感染细胞,sh-NC无关序列腺病毒感染细胞组为无关序列对照组,未感染腺病毒细胞组为空白组,应用PCR法检测AFAP1-AS1表达水平、CCK-8法测定细胞增殖情况、流式细胞术检测细胞凋亡情况,对比检测结果。结果:AFAP1-AS1表达水平检测结果显示:三种shRNA序列干扰效率均较无关序列对照组（sh-NC）强,差异有统计学意义（P＜0.05）;采用CCK-8法检测各组细胞凋亡情况,结果显示:经腺病毒sh3-AFAP1-AS1感染后,OCI-Ly1细胞系中实验组细胞吸光度较无关序列对照组和空白组显著降低,下调AFAP1-AS1可抑制GCB-DLBCL细胞的增殖（P＜0.05）;采用流式细胞仪检测各组细胞凋亡情况,结果显示:经sh3-AFAP1-AS1和sh-NC转染后,OCI-Ly1细胞实验组凋亡率明显高于无关序列对照组和空白组,下调AFAP1-AS1可诱导GCB-DLBCL细胞凋亡（P＜0.05）。结论:沉默GCB-DLBCL细胞中的AFAP1-AS1表达能有效抑制细胞增殖,诱导细胞凋亡,或可作为GCB-DLBCL治疗的靶目标。     

Venous thromboembolism in patients with diffuse large B-cell lymphoma

We conducted a retrospective record review to determine the frequency of venous thromboembolism (VTE) in patients with diffuse large B-cell lymphoma (DLBCL). All records from 1990 to 2001 of patients with the diagnosis of DLBCL at a tertiary care hospital were reviewed. Those with transformation from low-grade lymphoma, central nervous system lymphoma, HIV-related lymphoma or with incomplete records were excluded. All episodes of symptomatic VTE confirmed by imaging studies that were either present at diagnosis or occurred during initial treatment were identified. VTE occurred in 27 of 211 patients (12.8%). Stage I disease was associated with a low risk, whereas a high international prognostic index score increased risk. Of patients with VTE, thrombosis was present at diagnosis in 37% and occurred during the first chemotherapy cycle in 22% and during the first three cycles in 82%. The median survival of patients with VTE was 1.04 years [95% confidence interval (CI) = 0.75 - 1.33] compared to 5.2 years (95% CI 1.8 - 8.6) for those without VTE (P = 0.038). We conclude that VTE is a frequent complication of DLBCL that occurs particularly at diagnosis and during initial therapy, and it is associated with a worse prognosis.     

miR-155在弥漫性大B细胞淋巴瘤预后预测中的应用价值

目的:探讨miR-155在弥漫性大B细胞淋巴瘤预后预测中的应用价值。方法:选取我院收治的120例弥漫性大B细胞淋巴瘤作为研究对象,采用qRT-PCR检测所有患者癌组织miR-155的相对表达水平,根据miR-155的表达水平将所有患者分成miR-155高表达组（n=72）和miR-155低表达组（n=48）,比较两组患者的临床病理资料、生存率,采用 Cox比例风险回归模型对弥漫性大B细胞淋巴瘤患者的预后进行单因素和多因素分析,并分析miR-155对弥漫性大B细胞淋巴瘤细胞增殖和迁移能力的影响。结果:miR-155高表达组患者结外侵犯比例显著高于miR-155低表达组（P＜0.05）;miR-155高表达组患者3年无进展生存率（29.2%）及总体生存率（40.3%）均显著低于miR-155低表达组（81.3%和83.3%）;单因素和多因素分析结果均显示miR-155表达水平是DLBCL无进展生存期和总体生存期的影响因素;miR-155低表达组细胞划痕愈合速度(0.53±0.04)显著低于对照组细胞（1.0±0.03）(P＜0.05),miR-155低表达组细胞在培养的3、4 d的吸光度值显著低于对照组（0.38±0.01 vs 0.56±0.03;0.56±0.02 vs 0.76±0.02）（P＜0.05）。结论:弥漫性大B细胞淋巴瘤患者的miR-155表达水平显著影响患者的预后,其可能机制是通过影响弥漫性大B细胞淋巴瘤细胞的增殖和迁移能力,提示miR-155可能是弥漫性大B细胞淋巴瘤新的和可靠的预后生物标志物,值得进一步深入研究。     

Evolution of R-CHOP therapy for older patients with diffuse large B-cell lymphoma

Non-Hodgkin’s lymphoma is the fifth most common malignancy in adults in the USA. This disorder is especially relevant in the elderly patient population, as the median age of patients with this disorder is 65 years. Almost half of these disorders in older patients are of a diffuse large B-cell (DLBCL) subtype. The therapy of DLBCL has undergone a renaissance in the past decade, with the addition of rituximab to standard regimens, such as cyclophosphamide– doxorubicin–vincristine–prednisone (CHOP). Over this time, there have been several large Phase III treatment trials in which the CHOP and rituximab-CHOP (R-CHOP) regimens have been prospectively compared, including three trials confined to the elderly patient population. In these trials, it has been demonstrated repeatedly that the addition of rituximab results in an improved outcome, with higher response rates and prolongation in parameters including progression-free, event-free, disease-free and overall survival. In addition, this regimen has been well tolerated, even in older patients. Based upon these data, the R-CHOP regimen has now been established as the standard for initial therapy of DLBCL in older patients with DLBCL. However, issues still remain with regard to the ideal schedule of R-CHOP administration, specifically the optimal number of cycles of therapy (six vs eight), as well as cycle length (14 vs 21 days).     

Clinical characteristics and outcome for hepatitis C virus-positive diffuse large B-cell lymphoma

Several previous studies have addressed the association between hepatitis C virus (HCV) infection and non-Hodgkin lymphoma (NHL), but there are few studies on HCV-related diffuse large B-cell lymphoma (DLBL). We conducted this retrospective study to investigate the distinctive clinical characteristics and outcome for HCV-positive DLBL. We compared the clinical characteristics and outcomes of 32 HCV-positive DLBL cases from nine Korean institutions with those of 371 HCV-negative DLBL cases. A higher percentage of HCV-positive DLBL cases were associated with old age (≥60) than HCV-negative DLBL cases at diagnosis (59.4% vs. 36.1%, respectively, P = 0.009) and HCV-positive cases were less likely than HCV-negative cases to have extranodal involvement (53.1% vs. 71.1%, respectively, P = 0.044). The nodal presentation was the only independent factor favorably influencing the event free survival (EFS) in HCV-positive DLBL (HR = 0.11, 95% CI; 0.01 - 0.95, P = 0.012). In comparison to patients with HCV-negative DLBL, HCV-positive DLBL patients had a superior complete response rate (P = 0.023) and EFS (P = 0.02). In Korean patients, HCV-positive DLBL is more common with old age and has less extranodal involvement than does HCV-negative DLBL. The superior survival outcome for HCV-positive DLBL should be verified by further investigation, especially with respect to its correlation with transformed low-grade NHL.