microRNA-224在弥漫大B细胞淋巴瘤的表达及作用

目的研究miR-224在弥漫大B细胞淋巴瘤(DLBCL)患者中的表达与临床病理特征的关系,观察miR-224对弥漫大B淋巴瘤细胞系OCI-LY10增殖、侵袭能力的影响。方法收集86例DLBCL组织,并以22份无肿瘤细胞侵犯的正常淋巴结组织标本为对照,采用real-time PCR方法检测miR-224的相对表达水平,分析miR-224表达水平与患者临床病理特征的关系。对弥漫大B淋巴瘤细胞系OCI-LYl0进行miR-224反义寡核苷酸(ASO)转染并培养,MTT法检测细胞增殖能力,Transwell小室检测细胞侵袭能力,real-time PCR方法与Western blot方法检测细胞中bcl-2的表达水平。结果DLBCL患者淋巴瘤中miR-224表达水平显著低于正常淋巴结组织(P0.05),miR-224在DLBCL患者中的表达与患者年龄、性别、疾病分期均无相关性(P0.05)。miR-224ASO明显升高OCI-LYl0细胞的增殖侵袭能力,显著升高bcl-2蛋白与m RNA表达水平(P0.05)。结论miR-224抑制OCI-LY10细胞增殖侵袭可能与下调bcl-2蛋白表达相关。     

弥漫大B细胞淋巴瘤中LMO4的表达及意义

目的探讨LMO4在弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma, DLBCL)中的表达及其临床意义。方法应用免疫组化EnVision法检测123例DLBCL组织及60例反应性淋巴组织增生(reactive lymphoid hyperplasia, RLH)组织中LMO4蛋白的表达。采用免疫细胞化学和Western blot法分别检测LMO4蛋白在DLBCL细胞株LY-10、SUDHL-4以及正常人外周血淋巴细胞中的表达。结果免疫组化结果显示LMO4蛋白在DLBCL中的高表达率明显高于RLH组织(66.7%vs 23.3%,P0.05),其表达与肿瘤原发位置、免疫分型、IPI评分及Ann Arbor分期均有相关性(P0.05),而与其他临床病理特征无明显相关性。免疫细胞化学和Western blot均显示LMO4在DLBCL细胞株LY-10和SUDHL-4中呈高表达,在正常人外周血淋巴细胞中呈低表达。结论 DLBCL组织和细胞株中LMO4均呈高表达,在RLH和正常人外周血淋巴细胞中低表达,提示LMO4在DLBCL发生、发展中可能起重要作用。     

弥漫大B细胞淋巴瘤中Prdx6的表达及其预后意义

目的 探讨过氧化物酶6(peroxiredoxin-6,Prdx6)在弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达及其预后意义.方法 回顾性分析286例DLBCL中Prdx6的表达及与临床病理特征和预后之间的关系.结果 Prdx6在DLBCL中的表达高于正常淋巴结...     

microRNA-191在弥漫大B细胞淋巴瘤患者中的表达及其作用机制研究

目的分析miR-191在弥漫大B细胞淋巴瘤(DLBCL)患者中的表达与临床病理特征的关系,观察miR-191对弥漫大B淋巴瘤细胞系OCI-LY10增殖、侵袭能力的影响。方法采用荧光实时定量PCR方法检测86例DLBCL患者肿瘤组织和22例正常淋巴结组织中miR-191的表达,分析miR-191表达水平与患者临床病理特征的关系。对弥漫大B淋巴瘤细胞系OCI-LY10进行miR-191反义核苷酸(ASO)转染并培养,MTT法检测细胞增殖能力,Transwell小室检测细胞侵袭能力,实时PCR方法与Western blot方法检测细转染miR-191ASO后各组细胞PLCD1的m RNA和蛋白的表达。结果 DLBCL患者淋巴结组织中miR-191表达水平显著高于对照组,两者比较差异有统计学意义(P<0.05)。miR-191在DLBCL患者中的表达与患者年龄、性别无关(P>0.05),但是与疾病分期显著相关(P<0.05)。与对照组相比,miR-191 ASO组OCI-LYl0细胞的增殖侵袭能力明显降低,PLCD1蛋白与mRNA表达水平显著升高(P<0.05)。结论 miR-191在DLBCL中表达上调,其促进OCI-LY10细胞增殖侵袭的作用机制可能与下调PLCD1蛋白表达有关。     

PLK1和DNAPKcs在弥漫大B细胞淋巴瘤中的表达及临床意义

目的探讨PLK1和DNAPKcs在弥漫大B细胞淋巴瘤(DLBCL)中的临床意义及对DLBCL细胞增殖的影响。方法收集50例DLBCL患者的临床资料,并通过对其病理组织标本进行免疫组化检测PLK1和DNAPKcs的表达水平,分析它们的表达相关性及与DLBCL临床病理特征之间的关系。通过siRNA干扰PLK1和DNAPKcs,采用EdU实验观察DLBCL细胞增殖能力的变化。敲减PLK1的表达,Western blot实验检测DLBCL细胞DNAPKcs的表达变化。结果 PLK1和DNAPKcs表达水平在DLBCL病人和两种细胞系FARAGE和OCI-Ly3中显著高于正常人外周血单核细胞(P0.05),促进DLBCL肿瘤细胞的增殖,与Ann Arbor分期、患者是否出现B症状、IPI评分正相关(P0.05),与DLBCL患者年龄、性别无相关性;敲减PLK1后DNAPKcs的表达水平显著降低。结论 PLK1和DNAPKcs可作为DLBCL诊断和预后判断的重要指标。     

弥漫大B细胞淋巴瘤相关抗原特异TCR基因修饰T细胞TCRζ链基因的表达

目的:分析DLBCL相关抗原特异TCR基因修饰T细胞经肿瘤细胞刺激活化前后TCRζ链基因表达水平的变化.方法:利用SYBR Green Ⅰ荧光定量PCR,相对定量检测TCR基因修饰T细胞活化前后TCRζ链基因的表达情况,β2微球蛋白基因(β2M)为内参照,根据相对定量公式:2-ΔCt ×100%和2-ΔΔCt,计算TCR基因修饰T细胞活化前后TCRζ链基因的相对表达水平及其差异情况.结果:与未转染T细胞的TCRζ基因mRNA表达量(1.74±0.28)%相比,TCR转基因修饰T细胞TCRζ基因mRNA的表达水平没有发生明显的变化,表达量为(1.78±0.22)%;而经效/靶细胞(TCR基因修饰T细胞/Toledo细胞)混合培养后,活化的TCR基因修饰T细胞的TCRζ基因mRNA表达水平为(11.54±1.98)%,明显高于未刺激培养之前的TCR基因修饰T细胞和未转染T细胞的TCRζ基因的表达水平(P＜0.05),分别增长了(6.59 ±0.80)倍和(6.48±0.36)倍.结论:TCR转基因修饰T细胞受到抗原的刺激而活化,TCRζ链基因的表达相应增加.     

弥漫大B细胞淋巴瘤中CD27的表达及临床意义

目的探讨弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma, DLBCL)中CD27的表达及其临床意义。方法采用免疫组化EnVision法检测143例DLBCL组织中CD27蛋白的表达;应用FISH技术检测DLBCL组织中MYC、BCL-2、BCL-6基因重排情况。结果 143例DLBCL中,CD27蛋白阳性者46例,阳性率为32.2%。CD27阳性组中BCL-2重排阳性率(17.4%)明显高于CD27阴性组(4.1%),差异有统计学意义(P<0.01)。CD27阳性组病死率(30.4%)明显高于CD27阴性组(15.5%),差异有统计学意义(χ²=4.326,P=0.038)。CD27阳性者与阴性者的Kaplan-Meier生存曲线差异有显著性(χ²=4.485,P=0.034),阳性组生存期较短。结论 CD27高表达与DLBCL预后密切相关,可作为临床预后评价的指标之一。     

Comparative clinicopathological study of primary CNS diffuse large B-cell lymphoma and intravascular large B-cell lymphoma

Primary CNS diffuse large B-cell lymphoma (CNS DLBCL) is confined to the CNS, and constitutes a distinct entity. In the present study a series of 40 Japanese patients with CNS DLBCL who presented with neurological, but not systemic symptoms, was reviewed. Median survival was 18.7 months. CD5, CD10, Bcl-6, MUM-1, and Bcl-2 were positive in 30%, 10%, 84%, 100%, and 93% of patients, respectively. All CD10-negative patients had non-germinal center B-cell type. There was no significant difference in survival among the immunophenotypic subgroups. CNS DLBCL appeared to be homogenous as a group, which prompted the comparison with another distinct extranodal entity, intravascular large B-cell lymphoma (IVLBCL) in Japanese patients. CNS DLBCL patients did not differ in age, sex, or immunophenotype, including CD5 positivity, from IVLBCL patients, but were significantly less likely to have poor prognostic parameters than IVLBCL patients: the international prognostic index score was low or low–intermediate in 86% of CNS DLBCL patients and high or high–intermediate in 98% of IVLBCL patients. Notably, despite this difference, their survival curves almost overlapped. The present study highlights the issue of clinical distinctiveness of aggressive extranodal lymphomas, the peculiar migration and localization of which should be further clarified.     

Beclin 1 expression predicts favorable clinical outcome in patients with diffuse large B-cell lymphoma treated with R-CHOP

Beclin 1 plays a critical role not only in autophagy, but also in apoptosis and differentiation. The prognostic significance of beclin 1 in diffuse large B-cell lymphoma is largely unexplored. Immunohistochemical protein expression of beclin 1 in 118 tumor specimens from patients newly diagnosed with diffuse large B-cell lymphoma treated with rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone regimen from 2003 to 2007 was analyzed. Beclin 1 expression was observed in 101 (85.6%) patients. Low beclin 1 expression was related to B symptoms (P = .018), advanced Ann Arbor stage (P = .009), International Prognostic Index of 2 or higher (P = .049), elevated serum lactate dehydrogenase level (P = .037), and poor outcome using the revised International Prognostic Index model (P = .013). The complete remission rate after standard treatment was higher in patients with high beclin 1 expression than in those with low beclin 1 expression (77.5% versus 55.3%, P = .013). Beclin 1 expression was inversely associated with bcl-2 expression (P = .019) and positively associated with longer overall survival (P = .035) and progression-free survival (P = .013). In multivariate analysis, beclin 1 expression and the revised International Prognostic Index model independently predicted survival. The proposed clinicopathologic prognostic model including these 2 independent prognostic factors allowed classification of patients into 3 risk groups (P < .0001, respectively). Beclin 1 expression predicts superior clinical outcome in patients with diffuse large B-cell lymphoma treated with standard treatment. The novel prognostic model which divides patients with diffuse large B-cell lymphoma into different risk categories may help guide treatment planning.     

Fluorescent nanoparticle-mediated semiquantitative MYC protein expression analysis in morphologically diffuse large B-cell lymphoma

The current World Health Organization (WHO) classification defines a new disease entity of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements, making fluorescence in situ hybridization (FISH) screening for these genes mandatory. In addition, the prognostic significance of MYC expression was reported, with a cut-off value of 40%. However, interobserver discrepancies arise due to the heterogeneous intensity of MYC expression by immunohistochemistry. Moreover, a cut-off value of positivity for MYC protein in diffuse large B-cell lymphoma (DLBCL) varies among studies at present. Here, we applied a high-sensitivity semiquantitative immunohistochemical technique using fluorescent nanoparticles called phosphor-integrated dots (PID) to evaluate the MYC expression in 50 de novo DLBCL cases, and compared it with the conventional diaminobenzidine (DAB)-developing system. The high MYC expression detected by the PID-mediated system predicted poor overall survival in DLBCL patients. However, we found no prognostic value of MYC protein expression for any cut-off value by the DAB-developing system, even if the intensity was considered. These results indicate that the precise evaluation of MYC protein expression can clarify the prognostic values in DLBCL, irrespective of MYC rearrangement.     

Gene expression patterns in AIDS versus non-AIDS-related diffuse large B-cell lymphoma

Diffuse large B-cell lymphoma (DLBCL) is more prevalent and more often fatal in AIDS patients compared to immune-competent individuals. Potential explanations for these differences include distinct tumorigenic mechanisms and/or altered cellular microenvironments. We previously discovered that the TCL1 (T-cell leukemia-1) proto-oncogene is expressed in a high proportion of AIDS-DLBCL compared to DLBCL cases and that aberrant TCL1 expression causes DLBCL in a new transgenic mouse model. Here, we continue to search for other genes that may contribute to the differential pathogenesis of DLBCL in AIDS. Gene subtraction yielded over 1800 potential AIDS-DLBCL candidates, of which about 50% were unknown and not further considered. The remaining 50% of genes were annotated and, when combined with miniarray screening from multiple patient samples, were reduced to 18 candidate genes for extended analysis. These 18 genes showed distinct patterns of expression in both AIDS-DLBCL and DLBCL samples. However, unlike TCL1, none of these genes was preferentially associated with either AIDS-DLBCL or DLBCL. Our data suggest that the increased incidence and severity of AIDS-DLBCL compared to DLBCL is likely due to crippled immune surveillance rather than to markedly different gene expression profiles.     

Recurrent diffuse large B-cell lymphoma presenting initially as hemophagocytic syndrome

Hemophagocytic syndrome (HPS) is an extremely rare and life-threatening abnormality, and the cases secondary to B cell lymphoma are rare. We report a case of relapsed diffuse large B-cell lymphoma initially presenting with hemophagocytic syndrome. The patient developed multiple erythematous macules and progressive thrombocytopenia during the treatment, and died two weeks after admission. The HPS presented as an initial manifestation of the relapsed diffuse B-cell lymphoma and the maculea that appeared during the treatment might be a strong predictor of unfavorable outcome.     

CD3-positive diffuse large B-cell lymphoma relapses as CD3-negative large B-cell lymphoma: Loss of aberrant antigen expression in B-cell lymphoma after chemotherapy

Aberrant expression of CD3 on diffuse large B-cell lymphoma (DLBCL) is rare, and its mechanism and biological significance are currently unclear. Herein we report a case of Epstein-Barr virus-negative, CD3-positive DLBCL in a 53?year-old male, who had a remote history of renal transplantation. After standard chemotherapy, the patient was in clinical remission. He relapsed three years later, but at this time with apparent loss of CD3 expression. PCR-based IGK gene rearrangement studies demonstrated clonal amplicons with an identical nucleotide size between the primary and secondary DLBCL, confirming the clonal relationship despite their phenotypic differences. To our knowledge, this is the first case of CD3-positive DLBCL that demonstrated a loss of aberrant CD3 on relapse. The chronologic change in phenotype seen in this case suggests that the source of the patient’s lymphoma relapse may arise from either a quiescent subclone without CD3 expression, or from an upstream neoplastic precursor cell.     

Detection of germinal center B-cell lymphoma in archival specimens: critical evaluation of Bcl-6 protein expression in diffuse large B-cell lymphoma of the tonsil

Expression of Bcl-6 and CD10, markers for the tumor of the germinal center (GC) B-cell derivation, has been studied in primary diffuse large B-cell lymphomas (DLBCLs) of the lymph node, gastrointestinal tract, and mediastinum. In these studies, the coexpression rate of CD10 and Bcl-6 was relatively constant at 30% approximately 40%, but the frequency of Bcl-6+ tumors varied from 55% to 100%, raising doubts about the usefulness of Bcl-6 expression in identifying the tumor of GC B-cell derivation. Because the expression of Bcl-6 in tumors of non-GC B-cell origin has recently been reported, we critically evaluated the expression of Bcl-6 and CD10 in primary DLBCLs of the tonsil, a relatively common tumor in Japan and Korea. The cases (n = 51) represented a consecutive series for any recent 2-year period at several teaching hospitals in Korea and Japan. Formalin-fixed, paraffin-embedded specimens were used for immunostaining. Staining for Bcl-6 and CD10 was positive in 44 (86%) and 22 cases (45%), respectively. However, among those positive for Bcl-6 (>10% Bcl-6+ tumor cells), 2 basic patterns were recognized: uniform and nonuniform. The uniform pattern was characterized by a dense population (>75%) and a consistent density in any given area, resembling the staining pattern observed in GC or follicular lymphoma (FL) (the "GC/FL" pattern). In contrast, the nonuniform pattern exhibited a varying density from area to area, as well as a less-dense population (<75%). The uniform pattern was observed in 26 cases (51%). All but 1 (95%) of the CD10+ tumors coexpressed Bcl-6, with most (82%) displaying the uniform pattern. We conclude that tumors showing a uniform pattern of Bcl-6 expression should be distinguished from those showing a nonuniform pattern, because the former most likely represent tumors of GC B-cell derivation and the latter most likely represent tumors of non-GC derivation. GC B-cell lymphoma thus defined accounted for 51% of tonsillar DLBCL, a proportion comparable to that of the nodal DLBCL. CD10 expression correlated with the "GC/FL" pattern, but appeared to be not essential for the identification of GC B-cell lymphoma. This study suggests that a significant proportion of tonsillar DLBCLs in Asia is of GC B-cell origin rather than of mucosa-associated lymphoid tissue origin. This finding may have significance for clinical management of these lymphomas.     

One patient with double lymphomas: simultaneous gastric MALT lymphoma and ileal diffuse large B-cell lymphoma

Multiple different lymphomas in a single person are very rare. The author herein reports the case of a 69- year-old Japanese woman with double gastrointestinal lymphoma. The patient presented with epigastralgia. Endoscopic examination revealed erosions and elevation of the gastric body and a large ulcerated tumor of the terminal ileum. Biopsies were obtained from these lesions. The gastric lesion was MALT lymphoma with monocytoid B-cell proliferation and lymphoepithelial lesions. Light chain restriction was present. Helicobacter pylori were present on Giemsa stain. The gastric lesions did not regress despite of therapy, which were confirmed by follow-up biopsy. The ileal lesion was obvious diffuse large B-cell lymphoma. The lesion regressed by chemotherapy. The patient is now alive 3 years after the first presentation.