结外NK/T细胞淋巴瘤（鼻型）microRNA的表达研究

目的 通过分析结外NK/T细胞淋巴瘤（鼻型）肿瘤组织中microRNA( miRNA)的表达情况,探索其分子改变特征。方法 对1例结外NK/T细胞淋巴瘤（鼻型）和1例炎性鼻黏膜组织应用TaqMan低密度芯片分析665种miRNA的表达差异,发现95种miRNA存在变化。结合文献复习和数据库检索,从其中筛选差异明显、重要的8种miRNA。应用即时定量聚合酶链反应方法进一步对15例结外NK/T细胞淋巴瘤（鼻型）与3例炎性鼻黏膜组织验证这8种miRNA的表达情况。结果 8种miRNA验证结果与表达谱结果对比表达趋势一致,经统计学处理,其中3种重要miRNA[miR-223和miR-886-3p在结外NK/T细胞淋巴瘤（鼻型）组织中表达上调,miR-34c-5p表达下调]表达差异有统计学意义（P值分别为0.002、0.010和0.017）。结论 造血系分化发育、细胞增殖凋亡有关的miRNA：miR-223、886-3p和34c-5p在结外NK/T细胞淋巴瘤（鼻型）表达有明显异常,其是否参与肿瘤分子遗传学的改变及其意义值得深入探讨。     

NK/T cell lymphoma involving mediastinum: report of a case and review of literature

Extranodal natural killer (NK)/T-cell lymphoma is a very aggressive malignant neoplasia with a poor prognosis. Herein we reported a case of NK/T cell lymphoma involving mediastinum. It was a 28-year-old Chinese male patient. The tumor cells were medium-sized, had irregularly folded nuclei, and inconspicuous or small nucleoli with coagulative necrosis. The tumor cells were positive for CD3ε, TIA-1, but negative for CD56. In situ hybridization revealed that tumor cells also expressed Epstein-Barr virus encoded RNA. To our knowledge, this is the first case of NK/T cell lymphoma involving mediastinum.     

Primary bone natural killer/T cell lymphoma,nasal type without EBV infection: a case report

Primary bone NK/T cell lymphoma is very rare. We report a case of 52-year-old man of primary bone NK/T cell lymphoma and then progressed to NK leukemia. The patient had low-grade fever for 4-month, and Ultrasonic B revealed a diffuse hepatosplenomegaly without lymphadenopathy. PET scanning showed increased FDG uptake in many bones of the whole body. The diagnosis was established by bone specimen. These neoplastic cells demonstrated a typical immunophenotype of CD56, CD3, CD2 and MPO positive, and CD5, CD20, CD30, PAX-5, CD4 and CD8 negative. Primary bone ENKTL is very rare; it should be made with the combination of clinical feature, PET-CT image, and pathological characteristics, and should be distinguished from other lymphomas or leukemia involved in bone.     

结外鼻型NK/T细胞淋巴瘤28例临床病理分析

目的：探讨结外鼻型NK/T细胞淋巴瘤( extranodal natural killer/T-cell lymphoma, nasal type, EN-NK/TCL)的临床病理学特征、诊断及鉴别诊断。方法应用免疫组化、EBER原位杂交技术检测28例EN-NK/TCL,依据WHO(2008)淋巴造血组织肿瘤分类进行临床病理学分析。结果28例EN-NK/TCL中,男性13例,女性15例,男女之比为1∶1．2,平均年龄46岁,中位年龄44．5岁。18例发生于鼻腔,5例发生于皮肤,扁桃体、上腭、舌根、肾上腺、胃各1例。临床主要表现为鼻塞,皮肤病变可表现为皮疹、溃疡、斑块或结节形肿物。可伴有B症状、淋巴结肿大,可累及多个部位。肿瘤细胞为小、中、大淋巴细胞或呈混合型,常见血管中心性或血管破坏的生长方式及坏死,多伴有炎细胞浸润。免疫表型：T细胞标志物( CD3ε最敏感)、CD56及细胞毒性标志物均阳性,原位杂交EBER呈阳性。10例获得随访,其中1例死亡。结论 EN-NK/TCL好发于上呼吸道,其次是皮肤。结合形态学、免疫表型及EBER原位杂交结果可确诊。     

Extranodal NK/T‐cell lymphoma,nasal type of the uterine cervix: A case report

We report a rare case of extranodal NK/T‐cell lymphoma, nasal type of the uterine cervix that showed cytologic features mimicking cervical cancer. A 65‐year‐old woman presented with vaginal bleeding. Gynecological examination revealed a bulky tumor of the cervix. A conventional Papanicolaou‐stained cervical smear showed hypercellularity consisting of numerous variably sized cohesive clusters that mimicked epithelial tumors, with a necrotic and inflammatory background. A small number of individually scattered cells were also identified. These scattered cells showed pleomorphic, often cleaved, or horseshoe‐shaped nuclei and pale cytoplasm. Biopsy specimens revealed a diffuse growth of atypical cells with an angiocentric pattern. Extensive necrosis and infiltration of inflammatory cells were present. There were numerous mitotic figures. The tumor cells were positive for CD45RO, CD3ε, CD56, granzyme B, TIA‐1, CD7, and Epstein–Barr virus (EBV)‐encoded small RNA (EBER) by in situ hybridization, and negative for cytokeratin, chromogranin A, synaptophysin, CD4, CD5, CD8, CD20, and CD30. Based on these findings, this tumor was diagnosed as extranodal NK/T‐cell lymphoma, nasal type of the uterine cervix. Diagn. Cytopathol. 2016;44:430–433. © 2016 Wiley Periodicals, Inc.     

A rare pediatric case of a thymic cytotoxic and lymphoblastic T/NK cell lymphoma

Attempts to establish a concise classification of lymphoblastic lymphomas (LBLs) have gained momentum in recent years, mainly due to the expanding possibilities of immunohistochemical and genetic characterization of different disease entities. Thus, cases of immature lymphoid malignancies with unusual immunopathological features have been reported during the last years, suggesting the need for new LBL classification concepts. To further characterize and demonstrate the extended spectrum of LBL, we present an unusual pediatric case of LBL that could not be categorized into one of the subgroups and exhibited a benign course after surgical treatment and subsequent chemotherapy. A mediastinal tumor of a 6-year-old Caucasian boy was examined by clinical staging, light microscopy, immunohistochemistry and PCR assays. The tumor cells reacted with TdT and had a positive cytoplasmic immunoreactivity forCD3. Further T- aand NK-cell markers CD1a, CD4, CD8, CD10, and CD56 reacted positively, but CD57, CD16 and CD 30 (Ber H2) were all negative. CD34 as a marker for bipotential B/T-precursors was also positive. B cell markers (CD20, CD22, Cd79a and IgM) were all negative. No clonal B cell Ig or T cell gamma chain rearrangements were detectable. Epstein Barr virus and other Herpes Virus DNA were not detected using a sensitive PCR assay. The applied chemotherapy was tolerated well and a complete remission of the tumor was achieved (observation period three years after the initial diagnosis). Localization, morphology, and the expressions markers made the tumor a typical member of the LBL group. However, our case represents a rare pediatric lymphoma derived from a thymic precursor committed to T/NK-cell differentiation and a favourable outcome after chemotherapy.     

Extranodal NK/T-cell lymphoma, nasal type extensively involving the bone marrow

Extranodal NK/T-cell lymphoma, nasal type, is an aggressive EBV-associated lymphoma that mainly involves the nasal cavity but has also been reported to involve other extranodal sites without nasal involvement. In contrast to aggressive NK cell leukemia (a marrow-based aggressive leukemia of NK-cell origin); extensive bone marrow and blood involvement is extremely uncommon by nasal type NK/T lymphoma. We report a patient with extranodal NK/T-cell lymphoma, nasal type that developed extensive bone marrow involvement during the course of her disease with some overlapping features with aggressive NK-cell leukemia.     

Three cases of extranodal NK/T-cell lymphoma of the nasal type diagnosed by nasal brush cytology

Extranodal natural killer (NK)/T-cell lymphoma of the nasal type is a rare type of malignant lymphoma that is most common in Asian countries. Here we describe cytomorphologic, immunocytochemical, and molecular cytochemical features of three cases of NK/T-cell lymphoma of the nasal type diagnosed by nasal brush cytology. Cytomorphologic findings common among the three cases included the presence of several cell types, including nasal cavity epithelial cells, histiocytes, phagocytic histiocytes, and lymphoid cells, within a necrotic background. Suspected lymphoma cells were medium to large lymphoid cells possessing light blue and abundant cytoplasm. A characteristic feature of these cells was the presence of the tongue-like projections of cytoplasm from one or both sides of the cells. We believe these intriguing cytologic findings are indicators of NK/T-cell lymphoma of the nasal type. Azurophilic granules were observed in all cases, ranging from extremely fine granules to large granular lymphocyte (LGL)-like granules. Immunocytochemical and molecular cytochemical analyses showed staining for natural killer cell antigen CD56 as well as cytotoxic granule-associated proteins granzyme B7 (GrB7) and T-cell-restricted intercellular antigen-1 (TIA-1). EBV (Epstein-Barr virus) encoded small RNAs (EBER) positivity was shown by in situ hybridization, and no rearrangement of the TCRgamma gene was observed. Comparison between cytobrush and cotton swab methodology showed that cytobrush resulted in more cell-rich specimens than did cotton swabs, suggesting that nasal brush cytology with cytobrush is most useful in the diagnosis of NK/T-cell lymphoma of the nasal type.     

穿孔素与颗粒酶B在鼻NK/T细胞淋巴瘤诊断中的意义

目的　检测鼻NK/T细胞淋巴瘤中的穿孔素、颗粒酶B表达分布情况 ,为临床治疗和预后判断提供依据。方法　运用免疫组化S P法检测 11例鼻NK/T细胞淋巴瘤中穿孔素、颗粒酶B的表达和分布情况 ;同时与组织芯片中其他类型淋巴瘤进行比较研究 ,10例淋巴组织反应性增生作为对照。结果　 11例鼻NK/T细胞淋巴瘤中均见穿孔素和颗粒酶B过度表达。以 6 0例淋巴瘤制成组织芯片的 4 2例B细胞淋巴瘤中见 11例少许穿孔素和 14例少许颗粒酶B阳性表达细胞散在分布于瘤细胞间 ,10例外周T细胞淋巴瘤中 8例少许表达穿孔素和 9例少许表达颗粒酶B。 2例间变性大细胞淋巴瘤见少许穿孔素和颗粒酶B阳性表达细胞。 2例霍奇金淋巴瘤阴性。NK/T细胞淋巴瘤的穿孔素和颗粒酶B表达程度与T、B淋巴瘤比较 ,差异有显著性 (P <0 0 1)。结论　穿孔素和颗粒酶B是鉴定活化的细胞毒性细胞的免疫标记物 ,可作为NK/T细胞淋巴瘤的诊断性标记物 ;其在外周T细胞淋巴瘤和B细胞淋巴瘤中的表达 ,反映了机体存在抗肿瘤的免疫反应机制。     

血管内NK/T细胞淋巴瘤的研究进展

血管内NK/T细胞淋巴瘤(intravascular NK/T cell lymphoma,IVNKTL)罕见,其特征性表现为肿瘤性淋巴细胞选择性生长于血管腔内.不同地域患者临床表现多样且缺乏特异性,因此临床诊断困难.IVNKTL具有病程进展快、伴EBV感染、好发于亚洲,且以中国人多发等特点.在肿瘤未累及到多器官前进行早期诊治可有效改善IVNKTL患者的预后.     

蛋白酶体抑制剂MG132对NK/T淋巴瘤细胞增殖、凋亡和细胞周期的影响

目的： 探讨蛋白酶体抑制剂MG132（Z-leu-leu-leu-CHO,三肽基乙醛）对NK/T淋巴瘤细胞的增殖和细胞周期分布的影响,研究蛋白酶体抑制剂MG132治疗NK/T细胞淋巴瘤的潜在应用价值。方法: 用蛋白酶体抑制剂MG132处理细胞,噻唑蓝［ 3（4,5二甲基噻唑2）2,5二苯基四氮唑溴盐,3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide,MTT］法检测细胞增殖抑制率,倒置显微镜观察细胞形态改变,流式细胞仪检测细胞周期分布和凋亡。结果: 1 μmol/L MG132作用24 h,HANK1细胞增殖抑制率为（57.72±7.44）%,而0.1 μmol/L MG132作用24 h,增殖抑制率仅为（3.98±0.07）%;MG132剂量大,增殖抑制率高;1 μmol/L MG132作用24 h后,HANK1细胞G1和G2期细胞分别为（72.33±3.44）%和（12.86±1.29）%,对照组为（63.63±2.29）%和（7.94±1.91）%,用药组G1和G2期细胞明显高于对照组（P<0.01,P<0.05）; 早期凋亡细胞和晚期凋亡细胞分别为（33.57±2.10）%和（16.66±0.47）%,而对照组仅为（7.18±0.82）%和（3.81±1.06）%,与对照组相比,凋亡率明显增高 (P<0.01,P<0.01)。结论: MG132抑制NK/T细胞淋巴瘤细胞增殖,使细胞周期G1和G2期阻滞,促进细胞凋亡,并存在剂量和时间依赖关系,蛋白酶体抑制剂MG132很可能成为治疗进展期NK/T细胞淋巴瘤的药物。     

结外鼻型NK/T细胞淋巴瘤c-kit基因突变分析

目的分析鼻型NK／T细胞淋巴瘤（N—NK／T—L）组织中c—kit基因突变和表达情况,探讨N．NK／T-L可能的特异分子标记物。方法研究组为36例N-NK／T-L,对照组包括11例同部位B细胞淋巴瘤（BCL）和5例颈部非特殊性外周T细胞淋巴瘤（PTCL）;采用免疫组化EnVision法对淋巴瘤进行c—kit产物CD117标记;采用巢式PCR法对31例N—NK／T—L、11例BCL和5例PTCL肿瘤组织c—kit基因11和17号外显子片段进行扩增,PCR纯化产物测序和序列分析。结果N—NK／T—L组织学表现为凝固性坏死,不同大小和异型的肿瘤性淋巴细胞呈破坏性生长浸润于炎性背景中。36例N．NK／T-L免疫组化表达阳性率为：CD45RO75．0％、CD3e72．2％、TIA83．3％、GranzymeB61．1％、CD5680．5％、CD3030．6％,而全部病例CD117阴性。对照组5例PTCL均为TIA1阳性,仅1例GranzymeB阳性,而11例BCL仅CD20和CD79et阳性,其余标记均阴性。GranzymeB表达与N-NK／T-L肿瘤细胞的异型性具有相关性（P〈0．05）。尽管所有病例CD117染色阴性,但c—kit基因11和17外显子扩增序列分析表明8例（8／31,25．8％）N-NK／T—L病例具有基因突变,其中分别有4例涉及11外显子和17外显子;对照组11例BCL和5例PTCL中分别有1例具有11外显子突变。所有突变均为碱基替代错义突变,热点涉及571、572和821位点。结论上述结果提示除组织学特征外,GranzymeB可能是更可靠的N-NK／T—L诊断和鉴别诊断指标。c-kit错义突变可发生于约1／4的N—NK／T—L病例中,该突变可能引起c-kit的功能失调。     

原发肠道NK/T细胞淋巴瘤10例临床病理分析

目的探讨原发肠道NK/T细胞淋巴瘤,鼻型(extranodal NK/T-cell lymphoma,nasal type,ENKTL)的临床病理学特征、免疫表型、治疗及预后。方法回顾性分析10例原发肠道ENKTL的临床病理资料,并复习相关文献。结果 10例原发肠道ENKTL的男女比为7∶3,中位年龄37.5岁。其中位于结肠6例,小肠3例,直肠1例。根据Lugano Staging系统分类:Ⅰ期1例,Ⅱ期7例,Ⅲ期2例。淋巴瘤国际预后指数(IPI)评分0~2。初发症状为腹痛、腹胀、腹泻等。初期症状不明显,但进展较快。从确诊到死亡2~21个月,平均9.5个月。镜下见肿瘤细胞弥漫浸润肠壁全层,炎症改变背景,常伴有周围血管破坏和凝固性坏死。免疫表型:肿瘤细胞表达细胞毒性蛋白(TIA-1、Granzyme B、perforin)、CD2(100%)、CD3ε(90%)、CD56(80%)、p53(60%)、CD30和LMP1(30%),不表达EBNA3A,90%病例Ki-67高表达。所有病例EBER原位杂交均阳性。结论原发肠道ENKTL属罕见的侵袭性淋巴瘤,早期诊断较难,需结合临床资料、病理学特征、免疫表型综合诊断。该病尽管临床分期属于早期、IPI评分低,但预后差。     

Nodal T/NK-cell lymphoma of nasal type: a clinicopathological study of six cases

Aims:  To investigate the clinicopathological features of six unusual cases of nodal CD56+ and Epstein–Barr virus (EBV)+ T/natural killer (NK)-cell lymphoma, a putative nodal counterpart of nasal NK/T-cell lymphoma (nodal T/NK-cell lymphoma of nasal type) in comparison with nasal NK/T-cell lymphoma with secondary lymph node involvement ( n  = 24) and peripheral T-cell lymphoma (PTCL) of cytotoxic molecule (CTM)+ and EBV+ type ( n  = 21).
Methods and results:  All cases of nodal T/NK-cell lymphoma of nasal type exhibited diffuse infiltration of pleomorphic medium-sized to large tumour cells, reminiscent of those in CTM+ EBV+ PTCL. The tumour cells had a typical phenotype of nasal NK/T-cell lymphoma: CD2+, CD3ε+, CD4−, CD5−, CD56+, T-cell intracellular antigen-1+, granzyme B+, perforin+ and EBV+. However, four of six cases demonstrated clonal T-cell receptor γ-gene rearrangement on polymerase chain reaction analysis, unlike nasal NK/T-cell lymphoma. Comparison of clinical parameters and overall survival among the three groups demonstrated only minor differences.
Conclusions:  Nodal T/NK-cell lymphoma may occupy the grey zone between extranodal nasal-type NK/T-cell lymphoma and nodal CTM+ PTCL in a spectrum of NK to T-cell lymphomas that are EBV+. The close relationship between NK/T-cell lymphomas and cytotoxic T-cell lymphomas was also substantiated.     

Tumor cell nuclear diameter and CD30 expression as potential prognostic parameter in patients with extranodal NK/T-cell lymphoma, nasal type

Extranodal natural killer/T-cell lymphoma, nasal type (nasal ENKTL) is a distinct clinicopathologic entity of lymphoid tumors with variable size and differentiation of tumor cells. Nasal ENKTL is related to infection of the tumor cells with Epstein-Barr virus (EBV) and virtually all cases contain monoclonal episomal EBV DNA and detectable EBV encoded small nuclear RNAs (EBERs). Several clinical factors are known for their relation to the prognosis, but histopathologic prognostic factors of nasal ENKTL have not yet been well established. We evaluated the prognostic value of the longest nuclear diameter of EBER+ tumor cells (NDTC) along with the result of CD30 expression. Twenty two patients with newly diagnosed nasal ENKTL were evaluated regarding clinicopathologic characteristics. NDTC was measured using a computerized image analysis system. The results were expressed as the mean diameter of ≥ 50 cells in a patient. Median of the mean NDTC of the patients was 7.32 μm (5.15-11.27). Patients with larger mean NDTC (≥ 7.35 μm) had a poorer event-free survival (EFS) than those with smaller mean NDTC (<7.35 μm; p = 0.024) and had a tendency of inferior overall survival (OS) (p = 0.08). Patients with CD30 expression had a inferior EFS (p = 0.018) and OS (p = 0.011) compared those without CD30 expression. The NDTC of EBV infected tumor cell and CD30 expression had relation to survival in the current exploratory analysis.     

Both c-Myc and Ki-67 expression are predictive markers in patients with Extranodal NK/T-cell lymphoma,nasal type: A retrospective study in China

Extranodal natural killer (NK)/T-cell lymphoma, nasal type (ENKTL) is an increasingly recognized disease entity of aggressive tumor progression. The objective of this study was to investigate the clinical and prognostic significance of c-Myc and Ki-67 expression in ENKTL. Immunohistochemistry for c-Myc and Ki-67 was performed on tissue sections from 53 patients diagnosed with ENKTL, and their correlation with clinicopathologic variables was assessed. We also made a comparison between c-Myc positive and negative ENKTL on proliferation index (PI); and analyzed relationship between expression of c-Myc and Ki-67 index. The survival was analyzed by Kaplan–Meier product-limit method. Positive expression of c-Myc was shown in 64.2% of the patients, and high expression of Ki-67 was found in 66%. The PI in c-Myc-positive tumor cell was significantly higher than that in c-Myc-negative ones (P = 0.005). The positive correlation between c-Myc expression and Ki-67 index was also found (r = 0.454, P = 0.001). Patients of c-Myc expression were shown to be associated with unfavorable overall survival (OS) and decreased progression free survival (PFS) (P = 0.000 and 0.013, respectively). High expression of Ki-67 was also shown to be correlated with worse OS and PFS (P = 0.014 and 0.016, respectively). And patients expression of c-Myc+/Ki-67 high had the worst OS and PFS (P = 0.002 and 0.027, respectively). c-Myc may play an important role in the carcinogenesis of NK/T cell lymphoma by promoting cell proliferation. Both c-Myc expression and Ki-67 high expression in ENKTL patients may be valuable indicators for predicting the survival of ENKTL patients.     

Composite primary breast diffuse large B-cell lymphoma and T lymphoblastic leukemia/lymphoma: report of a case and review of literature

We reported a rare case of composite diffuse large B-cell lymphoma and T lymphoblastic leukemia/lymphoma (T-LBL) in a 46-year-old woman with progressive enlargement of the breast lump. The patient initially sought care at a local hospital with a single left breast lump without any other physical examination findings. Histopathological analysis of which revealed a diffuse infiltration of tumor cells that were rich in cytoplasm with vesicular chromatin and prominent nucleoli. Further analysis of immunohistochemistry showed a cluster of neoplastic cells which express B-cell markers: CD19, CD20 (weak), CD79a, PAX5 and BCL-2, but negative for T-cell markers such as CD2, CD3, CD5 and CD7. PET-CT showed evidence of lymphadenopathy and splenomegaly, which may indicate lymphoma infiltration. Then a biopsy of bone marrow showed typical features of T-LBL. The aberrant terminal deoxynucleotidyl transferase (TDT) and cCD3 positive T-cell population that lack surface CD10 and CD19 were identified by flow cytometric immunophenotyping. Polymerase chain reaction analysis of the T-cell receptor gamma gene and IgH gene revealed a clonal rearrangement and confirming T-cell clonality. Fluorescence in-situ hybridization (FISH) revealed a deletion of the P53 gene in these T-neoplastic cells may indicate a bad outcome of such disease. Neither the large B-cells nor T-cells were positive for Epstein-Barr virus encoded RNA.