cyclinD1过表达对子宫颈鳞状细胞癌SiHa细胞增殖及上皮间质转化的影响北大核心CSCD

目的 探讨cyclin D1过表达对子宫颈鳞癌SiHa细胞增殖、分化的影响及其他信号分子的变化情况.方法 采用PCR法扩增cyclin D1基因全长,构建cyclin D1-pcDNA3.1质粒转染人乳头状瘤病毒16阳性的SiHa细胞,筛选cyclin D1稳定过表达细胞株.采用逆转录聚合酶链反应（RT-PCR）法和Western blot分别检测转染细胞cyclin D1 mRNA和蛋白表达.采用四甲基偶氮唑盐（MTT）比色法绘制细胞生长曲线,采用RT-PCR法和Western blot检测转染细胞CK7、E-cadherin、波形蛋白、Snail基因的mRNA和蛋白表达;采用RT-PCR法检测增殖分化相关基因CDK4、CDK2、p21、p27、cyclin E、Rb、E2F、E6/E7、Ki-67基因的mRNA表达水平.对细胞同步化处理,用RT-PCR法检测细胞周期不同时间点cyclin D1及p21基因的mRNA表达情况.结果 成功构建cyclin D1稳定过表达的G-3细胞株.生长曲线及Ki-67 mRNA升高（P〈0.05）提示G-3细胞增殖速度加快,G-3细胞中波形蛋白、Snail基因和蛋白表达明显增加（均P〈0.05）,E-cadherin、CK7基因和蛋白表达明显降低,提示转染细胞发生了上皮间质转化.cyclin D1过表达后,CDK4、CDK2、p21、p27、cyclin E表达增加,Rb、E2F、E6/E7、p16表达无明显改变,p21与cyclin D1表达趋势基本一致,在细胞周期不同时间点表达存在波动性.结论 转染诱导cyclin D1过表达可促进SiHa细胞增殖和上皮间质转化,这一过程中伴随着CDK4、CDK2、p21、p27、cyclin E基因表达的上调.cyclin D1过表达时,p21表达量也增高,可能通过影响波形蛋白表达参与了对SiHa细胞上皮间质转化的调控.     

细胞周期蛋白DI表达、聚合酶链反应及荧光原位杂交检测t（11；14）（q13；q32）易位在套细胞淋巴瘤诊断中的应用

细胞遗传学和分子遗传学的研究证实，70%-80%套细胞淋巴瘤（MCL）中可以检测到t(11:14)(q13:q32)，易位使位于11q13上的bel-1癌基因（也称为PRADI，parathyroid adenomatosis，甲状旁腺腺瘤病或CCND1）处于14q32上1gH基因增强了的调控下，从而被激活，使其编码的细胞周期蛋白（cyclin）D1过度表达。这种易位在其他小     

套细胞淋巴瘤石蜡包埋组织中细胞周期蛋白D1和t(11;14)易位检测的临床病理意义

目的 探讨套细胞淋巴瘤石蜡包埋组织中细胞周期蛋白(cyclin)D1和t(11;14)易位检测的可行性及其诊断和鉴别诊断价值。方法 收集套细胞淋巴瘤36例,对照组小B细胞恶性淋巴瘤71例,均为石蜡包埋组织,运用免疫组织化学方法观察cyclin D1的表达;用半巢式聚合酶链反应(PCR)法检测t(11;14)易位,以看家基因β-肌动蛋白(actin)作为内对照检测DNA质量。结果 (1)36例套细胞淋巴瘤中26例(72．2％)表达cyclin D1,对照组无1例表达。(2)107例标本中101例(94．4％)可检出β-actin DNA表达。36例套细胞淋巴瘤中22例检出t(11;14)易位,对照组无1例检出。去除B-actin和t(11;14)易位均阴性2例,套细胞淋巴瘤中t(11;14)易位检出率为64．7％。(3)36例套细胞淋巴瘤中cyclin D1染色和(或)t(11;14)易位检测阳性病例为29例,总阳性率为80．5％。结论套细胞淋巴瘤石蜡包埋组织中cyclin D1和t(11;14)易位的检测具较高的特异性和可行性,两者的综合应用有助于正确的诊断和鉴别诊断。     

小B细胞恶性淋巴瘤形态学和免疫组织化学研究

目的：探讨各种小B细胞恶性淋巴瘤的形态学、免疫表型特征及其鉴别诊断。方法：对15例小淋巴细胞性淋巴瘤（SLL）、3例淋巴浆细胞性淋巴瘤（LPL）、36例滤泡性淋巴瘤（FL）、25例套细胞淋巴瘤（MCL）、7例淋巴结边缘区B细胞淋巴瘤（MZL）和30例黏膜相关淋巴细胞型结外边缘区B细胞淋巴瘤（MALT－MZL）的石蜡切片进行HE形态学观察和CD5、CD10、CD23和cyclinD1等抗体的免疫组织化学分析。结果：各种小B细胞恶性淋巴瘤在组成细胞和组织结构上各具特征；免疫表型：SLL表达CD5（82％）和CD23（80％），FL表达CD10（87％），MCL表达cyclinD1（84％）和CD5（80％），MZL／MALT－MZL和LPL均不表达CD5、CD10、CD23和cyclinD1。结论：各种小B细胞恶性淋巴瘤均是独立疾病，各具形态学和免疫表型特征，结合HE形态学观察和CD5、CD10、CD23、cyclinD1等免疫组化分析有助于正确诊断和鉴别诊断。     

结外B细胞淋巴瘤组织中API2-MALT1融合基因的检测及其意义

目的了解API2-MALT1融合基因mRNA多种变异体在多个部位黏膜相关淋巴组织结外边缘区B细胞淋巴瘤（MALT淋巴瘤）、结外弥漫大B细胞淋巴瘤（DLBCL）以及桥本甲状腺炎中的分布特征,探讨t（11;18）（q21;q21）与上述淋巴瘤的临床病理特征和预后的关系及意义。方法收集手术切除的MALT淋巴瘤62例（肺10例、胃31例、肠9例及甲状腺12例）、DLBCL32例（胃16例、肠13例和甲状腺3例）及桥本甲状腺炎8例标本,通过逆转录聚合酶链反应（RT-PCR）检测所有病例的API2-MALT1 mRNA,5例淋巴结反应性增生作为阴性对照。根据检测结果将94例淋巴瘤分为API2-MALT1阳性及阴性组,比较两组的临床病理特征和生存率（随访6-120个月）。结果94例淋巴瘤中39例检出API2-MALT1 mRNA（MALT淋巴瘤28例,结外DLBCL 11例）。8例桥本甲状腺炎及阴性对照组均未检出融合基因。共检出A1446-M814和A1446-M1123两种变异体,以后者多见。融合基因mRNA检出率在甲状腺淋巴瘤最低,而在肺、胃肠较高。与阴性组相比,阳性组临床分期较早,浸润程度较轻、复发率较低,5年生存率较高。结论API2-MALT1融合基因mRNA在MALT淋巴瘤和结外DLBCL中都可被检出,而在桥本甲状腺炎中未检出。MALT1基因在1123bp断点较高的发生率可能是国人API2-MALT1形成时的一个特点;表达API2-MALT1融合基因的B细胞淋巴瘤具有更加惰性的临床过程和较好的存活情况,将其视为同一谱系更能体现其发展。     

SOX11可表达于Cyclin D1阴性母细胞样套细胞淋巴瘤

近期的研究表明SOX11表达有助于套细胞淋巴瘤(MCL)的诊断,包括具有典型形态学改变的cyclin D1阴性MCL。本文作者探讨了SOX11在B细胞非霍奇金淋巴瘤(B-NHL)某些亚型中的表达模式及其特异性,并将其作为免疫表型特征分析cyclin D1阴性的母细胞样MCL(该组病例系首次报道)的识别价值。作者应用免疫组化方法评估了     

原发淋巴结套细胞淋巴瘤临床病理分析

目的：探讨原发淋巴结套细胞淋巴瘤（MCL）的临床病理与免疫组化特点。方法：收集6例淋巴结MCL,免疫组化ABC法确定肿瘤细胞特征,使用的抗体有CD45、CD20、CD79、CD45RO、CD30、CD68、TdT、CD43、CD5、cyclinD1、c-myc,IgD,IgM等。结果：光镜可将MCL分为4种亚型：套区型1例,结节型1例,弥漫型2例,母细胞化型2例。肿瘤细胞表达全B细胞标记,IgD CD43 ,cyclinD1(5/6),CD5(4/6) 。结论：MCL是一种具有特殊免疫表型的B细胞淋巴瘤,不同的组织学构型其预后可能不同,临床应与其它类型B细胞淋巴瘤鉴别,如淋巴结边缘区B细胞淋巴瘤（MZL）,滤泡性淋巴瘤（FL）及CLL／SLL等鉴别。     

根据WHO淋巴造血系统肿瘤新分类对山西省淋巴瘤分布特点的分析

目的根据WHO淋巴造血系统肿瘤新分类标准、分析山西省恶性淋巴瘤的分布特点。方法重新阅读HE切片,选用免疫组织化学ABC法标记间变性淋巴瘤激酶（ALK）1、bcl-6、CD（1α、3、4、5、7、8、10,15、20、23、30、43、56、68、79α和99）、细胞周期蛋白（cyclin）D1、上皮膜抗原（EMA）、IgD,k,λ、潜伏膜抗原（LMP）1、PAX5、末端脱氧核苷酸转移酶（TdT）和Vs38C;原位杂交方法标记EBER RNA。按照WHO淋巴造血系统肿瘤新分类标准,对山西省肿瘤医院存档的447例淋巴瘤组织标本重新分类。结果447例淋巴瘤中,385例（86．1％）为非霍奇金淋巴瘤（NHL）,62例（13．9％）为霍奇金淋巴瘤（HL）。68．3％NHL为B细胞来源,30．6％为T和NK细胞来源,组织细胞来源的肿瘤仅占3例（0．8％）。弥漫大B细胞淋巴瘤（DLBCL）为最常见的类型（35．1％）,其他依次为外周T细胞淋巴瘤、非特殊型（PTun,12．0％）、黏膜相关淋巴组织结外边缘区B细胞淋巴瘤（MALT淋巴瘤,11．7％）,滤泡性淋巴瘤（FL,8．6％）,前体淋巴母细胞性淋巴瘤（T-LBL,7．0％）,间变性大细胞淋巴瘤（ALCL,4．2％）,小淋巴细胞性淋巴瘤（B-SLL,3．6％）和套细胞淋巴瘤（MCL,2．6％）。263例B细胞淋巴瘤105例（39．9％）表达免疫球蛋白轻链,包括52例K和53例λ。263例B细胞淋巴瘤14例表达LMP-1,14例表达EBER;119例T和NK细胞淋巴瘤6例表达LMP-1,19例表达EBER,NHL中LMP-1和EBER表达具有不一致性。62例HL37例（59．7％）一致表达LMP-1和EBER RNA,包括7例富于淋巴细胞型HL、11例混合细胞型HL和19例结节硬化型HL。结论所搜集到的山西省DLBCL的比率类似于美国、澳大利亚、日本和韩国,FL的比率明显低于美国和澳大利亚。     

黏液样/圆细胞型脂肪肉瘤石蜡包埋组织中FUS-CHOP融合基因检测的临床病理学意义

目的 探讨在甲醛固定、石蜡包埋组织中检测FUS-CHOP融合基因的可行性及其对黏液样／圆细胞型脂肪肉瘤的诊断和鉴别诊断的价值。方法 收集黏液样／圆细胞型脂肪肉瘤档案标本44例,以非典型性／分化良好型脂肪肉瘤、多形性脂肪肉瘤、低度恶性黏液纤维肉瘤／黏液样恶性纤维组织细胞瘤等作为阴性对照,共60例。所有标本均为甲醛固定、石蜡包埋组织。用嵌套式逆转录-聚合酶链反应(RT-PCR)方法检测FUS-CHOP融合基因mRNA表达并经测序证实,以看家基因PGK作为内对照检测mRNA质量。结果 104例肿瘤标本中,93例(89．4％)可检出PGK mRNA表达,其中黏液样／圆细胞型脂肪肉瘤标本PGK阳性39例(88．6％),对照组PGK阳性54例(90％)。44例黏液样／圆细胞型脂肪肉瘤中20例可检出Ⅱ型FUS-CHOP融合基因表达,未能检测到Ⅰ型FUS-CHOP融合基因表达,排除内对照阴性病例5例,阳性率为51．3％(20／39)。对照组60例肿瘤标本均未检出FUS-CHOP融合基因表达。结论 (1)嵌套式RT-PCR方法检测FUS-CHOP mRNA表达可用于甲醛固定、石蜡包埋组织。(2)FUS-CHOP是黏液样／圆细胞型脂肪肉瘤的特异性分子遗传学标志物。采用嵌套式RT-PCR方法检测FUS-CHOP敏感性较高,特异性强,可用于该肿瘤的诊断和鉴别诊断。     

bcl-10蛋白异常表达对黏膜相关淋巴组织结外边缘区B细胞淋巴瘤的诊断价值

目的探讨bcl-10蛋白表达对黏膜相关淋巴组织结外边缘区B细胞淋巴瘤（MALT淋巴瘤）的诊断价值。方法收集140例不同部位的MALT淋巴瘤,包括胃38例、眼眶35例、肠16例、皮肤15例、涎腺15例、肺14例、甲状腺3例、其他部位4例。对照：10例扁桃体反应性滤泡增生（RFH）、5例眼眶的淋巴组织增生和143例非MALT淋巴瘤、不同类型的非霍奇金淋巴瘤（NHL）,包括20例NK／T细胞淋巴瘤、20例滤泡性淋巴瘤（FL）、20例间变性大细胞淋巴瘤（ALCL）、20例淋巴结内弥漫大B细胞淋巴瘤（DLBCL）、10例原发胃DLBCL、13例淋巴结边缘区淋巴瘤（NMZL）、12例套细胞淋巴瘤（MCL）、11例脾脏边缘区淋巴瘤（SMZL）、6例血管免疫母细胞性T细胞淋巴瘤（AITL）、6例外周T细胞淋巴瘤（PTCL）、3例B．小淋巴细胞淋巴瘤（B-SLL）、1例淋巴浆细胞性淋巴瘤（LPL）和1例浆细胞瘤。免疫组织化学EnVision法检测bcl-10蛋白;免疫组织化学双标记法检测CD20与bcl-10的共表达。结果在扁桃体RFH中,bel-10蛋白呈中等强度表达于生发中心B细胞质中,套细胞不表达,边缘区细胞和副皮质区T细胞呈弱表达。在眼眶淋巴组织增生中,2例bel-10阴性,3例主要呈淋巴滤泡生发中心B细胞质阳性,与扁桃体RFH的表达类似。在非MALT淋巴瘤的其他类型NHL中,除3例（3／10）原发胃DLBCL呈胞核阳性外,其余均未见胞核表达;在不同NHL中的胞质阳性分别为：结内（12／20）和胃（7／10）DLBCL、FL和ALCL（16／20）、PTCL（5／6）、AILT（6／6）、NMZL（13／13）、SMZL（11／11）、B-SLL（3／3）和浆细胞瘤（1／1）,11例MCL呈胞质可疑阳性,20例NK／T细胞淋巴瘤和1例LPL阴性;在部分淋巴瘤中可见肿瘤性细胞表达而反应性小淋巴细胞不表达：MALT淋巴瘤之bcl-10的总表达率为92．1％（129／140）,其中54．3％（76／140）胞质阳性,37．9％（53／140）胞核阳性;但不同部位之胞核阳性率有所不同。在MALT淋巴瘤中,bcl-10蛋白核强表达最常见于眼眶（25．7％,9／35）;除出现异常bcl-10胞核表达外,约20％有反应性滤泡的病例呈生发中心失表达。双标记显示bcl-10阳性细胞为CD20阳性细胞,但CD20阳性细胞多于bcl-10阳性细胞。结论（1）淋巴细胞增生性病变中bcl-10蛋白普遍表达,细胞质表达可出现在多数NHL和反应性增生中,但在淋巴瘤中呈肿瘤细胞表达而反应性细胞不表达,提示bcl-10异常可能与部分淋巴瘤的形成有关;（2）细胞核内bcl-10异常表达主要见于MALT淋巴瘤;眼眶、肺等部位的胞核强阳性和生发中心阴性的特殊模式,对MALT淋巴瘤的诊断及其与反应性病变的鉴别诊断有一定辅助意义。     

Immunohistochemical detection of cyclin D1 using optimized conditions is highly specific for mantle cell lymphoma and hairy cell leukemia.

Mantle cell lymphoma (MCL) is more aggressive when compared with other lymphomas composed of small, mature B lymphocytes. Cyclin D1 is overexpressed in MCL as a result of the translocation t(11;14)(q13;q32). Cyclin D1 immunohistochemistry in fixed, paraffin-embedded tissue contributes to the precise and reproducible diagnosis of MCL without the requirement of fresh tissue. However, its use in bone marrow biopsies is not well established. In addition, increased levels of cyclin D1 mRNA have been found in hairy cell leukemia but have not consistently been detected by immunohistochemistry. We used a polyclonal antibody and heat-induced antigen retrieval conditions to evaluate 73 fixed, paraffin-embedded bone marrow, spleen, and lymph node specimens with small B-cell infiltrates, obtained from 55 patients. Cyclin D1 was overexpressed in 13/13 specimens of MCL (usually strong, diffuse reactivity in most tumor cells) and in 14/14 specimens of hairy cell leukemia (usually weak, in a subpopulation of tumor cells). No reactivity was detected in five cases of B-chronic lymphocytic leukemia; five cases of splenic marginal zone lymphoma; six cases of nodal marginal zone cell lymphoma; two cases of gastric marginal zone cell lymphoma; or ten benign lymphoid infiltrates in bone marrow, spleen, or lymph nodes. In summary, although the total number of studied cases is small and a larger series of cases may be required to confirm our data, we present optimized immunohistochemical conditions for cyclin D1 in fixed, paraffin-embedded tissue that can be useful in distinguishing MCL and hairy cell leukemia from other small B-cell neoplasms and reactive lymphoid infiltrates.     

bcl-1 REARRANGEMENT AND CYCLIN D1 PROTEIN EXPRESSION IN MANTLE CELL LYMPHOMA

Centrocytic lymphoma, or mantle cell lymphoma (MCL), is characterized by a chromosomal translocation t(11;14) (q13;q32) involving the bcl-1 locus on chromosome 11. Cyclin D1 is a cell-cycle regulatory protein essential for G1–S transition and has been identified as a potential transforming gene affected by the translocation. In this study, 32 cases of MCL were analysed for the bcl-1 rearrangement and cyclin D1 protein expression. In 17 cases, a rearrangement at the major translocation cluster of bcl-1 could be detected. Twenty-four cases exhibited nuclear cyclin D1 expression that was not detectable in other B-cell lymphomas ( n =40) or in normal B-cells. In nine MCL samples, cyclin D1 was expressed without a detectable bcl-1 rearrangement. The detection of a t(11;14) by means of classical cytogenetics in one of these cases, however, may suggest that this discrepancy could be due to chromosomal breakages outside the typical translocation cluster region. In two cases, a bcl-1 rearrangement was not accompanied by cyclin D1 expression. This study provides further evidence that cyclin D1 is involved in the pathogenesis of MCL and can be exploited as a diagnostic marker in the differential diagnosis of B-cell lymphomas and in the identification of MCL.     

Large cell variants of CD5+, CD23- B-cell lymphoma/leukemia

CONTEXT: Mantle cell lymphoma (MCL), and its leukemic phase, constitute a well-studied hematologic malignancy with known overall survival, prognostic indicators, morphologic findings at diagnosis and in bone marrow, and known incidence of the bcl-1 immunoglobulin gene rearrangement. Large cell variants of B-cell lymphoma/leukemia with a mantle cell immunophenotype (CD5+, CD23-), including but not limited to blastic MCL, prolymphocytoid MCL, blastic mantle cell leukemia, and prolymphocytic mantle cell leukemia, are not as well characterized. Although blastic MCL is known to be associated with a shorter overall survival than conventional MCL, the large cell variants of B-cell lymphoma/leukemia with a mantle cell immunophenotype have not been described as fully as conventional MCL. OBJECTIVE: The purpose of the present study was to describe the large cell variants of B-cell lymphoma/leukemia with a mantle cell immunophenotype. DESIGN: Nineteen cases of large cell variants of CD5+, CD23- B-cell lymphoma/leukemia are reviewed and described in regard to morphology, bone marrow morphological findings, Cyclin D1 immunostaining, and bcl-1 analysis. Clinical data were not available owing to the varied clinical sources of the specimens. SETTING: Tertiary-care academic institution. RESULTS: Lymph node involvement in blastic CD5+, CD23- B-cell lymphoma was diffuse (100%) with a nodular component (33%) or focal mantle zone pattern (10%). Bone marrow involvement in blastic CD5+, CD23- B-cell lymphoma was seen in only 27% of cases and was composed predominantly of small, slightly irregular lymphocytes. Cyclin D1 was demonstrated in 60% of the 15 cases analyzed and more sensitive in B5-fixed tissue. Bcl-1 (performed in 5 cases) was not detected in the 4 cases of blastic CD5+, CD23- B-cell lymphoma analyzed and was detected in the case of the prolymphocytoid MCL. Cyclin D1 was demonstrated in all 4 bcl-1 negative cases and was negative in the bcl-1 positive prolymphocytoid MCL. CONCLUSION: Careful analysis of clinical data, morphology, immunophenotype, Cyclin D1 expression, and molecular analysis are required to differentiate the unusual large cell variants of MCL from other processes.     

CD5-undetected by immunohistochemistry, t(11;14)(q13;q32)-positive conjunctival mantle cell lymphoma: a case report

Most primary ocular adnexal lymphomas are extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT). A few cases of ocular adnexal mantle cell lymphomas have been reported in the literature. We present a case of mantle cell lymphoma presenting as conjunctival mass. A 58-year-old man presented with a palpable mass in the left lower tarsal conjunctiva incidentally detected one month previously. Histopathologic examination showed proliferation of monomorphous small-to-medium sized lymphoid cells. On immunohistochemistry, tumor cells were positive for CD20, bcl-2, and cyclin D1, and negative for CD5. PCR analysis for immunoglobulin heavy chain gene rearrangement showed monoclonal B-cell proliferation. t(11;14)(q13;q32), involving the CCND1 and IGH genes, was detected in interphase fluorescent in situ hybridization using formalin-fixed, paraffin-embedded tissue; however, MALT1 gene translocation was not observed. The final diagnosis was mantle cell lymphoma. There was no lymphadenopathy; however, bone marrow involvement of the lymphoma was suspected. The patient has been receiving systemic chemotherapy. This case emphasizes the differential diagnosis of conjunctival mantle cell lymphoma from extranodal marginal zone B-cell lymphomas of MALT regarding the clinical and pathological aspects.