MALT淋巴瘤中bcl-10与NF-κB的表达及其意义探讨

目的研究在黏膜相关淋巴组织结外边缘区淋巴瘤（extranodal marginal zone Bcell lymphoma of mucosa associated lymphoid tissue，MALT淋巴瘤）中bcl-10核表达和NF-KB／p65活化的关系，探讨bcl-10核表达对MALT淋巴瘤发生的价值。方法收集不同部位MALT淋巴瘤74例（包括29例胃、16例肠、15例眼眶、8例肺、2例唾液腺、1例甲状腺、1例唇、1例胸腺和1例附睾），用免疫组化EnVision法检测bcl-10和NF—κB／p65的表达。结果在74例MALT淋巴瘤中，bcl-10的阳性率为94．6％（70／74），其中胞质阳性率为52．7％（39／74），胞核阳性率为41．9％（31／74）；NF—κB／p65的阳性率为100％（74／74），其中细胞质阳性率47．3％（35／74），细胞核和细胞质共同阳性率占52．7％（39／74）。统计结果显示bcl-10核阳性与NF—κB核表达间有显著相关性（P〈0．001）。结论在MALT淋巴瘤中，bcl-10核表达对NF—κB活化有显著影响，可能是促进细胞增生、导致肿瘤形成的重要因素。     

乳腺癌中bcl-2表达与p53和雌激素受体关系的研究

目的　研究乳腺癌中bcl- 2基因表达与p5 3基因和雌激素受体的关系以及对乳腺癌生物学行为影响。方法　应用S -P免疫组织化学方法对 4 5例乳腺癌石蜡包埋组织进行检测。结果在人的乳腺癌中bcl- 2、p5 3蛋白和雌激素受体阳性表达率分别为 5 5 5 %、5 1 1%和 4 6 7%。bcl- 2阳性表达与肿瘤大小、淋巴结是否发生转移无关 (P >0 0 5 ) ;与p5 3阳性表达呈负相关 (P <0 0 5 ) ;与乳腺癌恶性程度和雌激素受体阳性表达呈正相关 (P <0 0 5 )。结论　bcl- 2基因、p5 3基因和雌激素共同参与了乳腺癌的发生和发展 ;bcl- 2阳性表达常会减弱p5 3的表达 ;为综合评价bcl- 2基因在乳腺生物学行为中的作用提供了一些线索。     

细胞周期蛋白D1、Ki-67和bcl-2的表达与CD117阳性的胃肠道间质瘤生物学行为的关系

目的 检测细胞周期蛋白D1（cyclin D1）、Ki-67和bcl-2在胃肠道间质瘤（GIST）中的表达,探讨它们在GIST发生、发展中的作用及临床病理意义。方法 59例手术切除GIST标本进行CD117、CD34、平滑肌肌动蛋白（SMA）、结蛋白、S-100、cyclin D1、bcl-2和Ki-67免疫组织化学染色,同时进行病理形态学观察包括形态学类型、肿瘤大小、坏死和核分裂象。所有病例随访2～9年。所有数据进行单因素、多因素和相关分析。结果 随访40例患者一直健在,15例患者死于GIST,4例患者死于其他原因。统计学分析显示肿瘤直径〉5cm、有坏死、核分裂象在每50个高倍视野〉5个、Ki-67增殖指数（LI）〉5％、cyclin D1和bcl-2免疫组织化学染色强阳性都可以作GIST患者手术后的预测指标,且具有统计学意义;核分裂象和Ki-67增殖指数是独立的预测指标;Ki-67 LI≥5％和核分裂象≥5／50 HPF呈正相关（r=0．532,P〈0．01）;cyclin D1与bcl-2强阳性表达呈正相关（r=0.273,P〈0．05）。结论 肿瘤大小、坏死、核分裂象、cyclin D1、Ki-67增殖指数和bcl-2可作为GIST患者临床预测指标;核分裂象和Ki-67增殖指数可作为独立的预测指标;cyclinD1与bcl-2呈明显相关性,Ki-67免疫组织化学染色可以代替核分裂象作为一项很有用的预测指标。     

Bcl-2和ER在甲状腺乳头状癌中的表达及意义

目的：探讨细胞凋亡相关基因bcl-2和雌激素受体（ER）在甲状腺乳头状癌中的表达及意义。方法：用免疫组织化学sP法研究不同甲状腺组织中ER和bcl-2的表达，其中甲状腺乳头状癌50例、甲状腺良性腺瘤30例、腺瘤旁正常组织16例。结果：甲状腺乳头状癌组织中ER和bcl-2蛋白阳性率分别为54．0％（27／50）、66．0％（33／50），甲状腺良性腺瘤中ER和bcl-2蛋白阳性率分别为26．7％（8／30）、36．7％（11／30），正常甲状腺组织中ER阳性率为12．5％（2／16）、而bcl-2则不表达。甲状腺乳头状癌中ER的表达明显高于甲状腺良性腺瘤和正常甲状腺组织（P〈0．05），bcl-2的表达明显高于甲状腺良性腺瘤（P〈0．05）；且甲状腺乳头状癌中bcl-2和ER表达存在正相关关系（P〈0．05）。结论：雌激素在甲状腺乳头状癌的发生、发展中有促进细胞增殖的作用；对ER和bcl-2的联合检测在甲状腺乳头状癌的诊断及内分泌治疗有一定的临床意义。     

胃肠道间质瘤中PDGFRα基因突变的检测

目的检测胃肠道间质瘤（gastrointestinal stromal tumors，GIST）中PDGFRa基因的突变位点及类型，探讨其在GIST发病机制中的作用。方法运用PCR—SSCP法和DNA测序检测22例GIST标本中PDGFRaexon12和exon18的基因突变，并以3例平滑肌瘤、1例神经鞘瘤、20例正常胃肠道组织作为对照。结果22例GIST中共有5例发生了PDGFRα突变（22．7％），平滑肌瘤、神经鞘瘤和正常胃肠道组织中未检测出PDGFRα的突变。PDGFRa的突变在良恶性、性别、年龄、肿瘤部位、大小及组织学分型之间的差异均无显著性。结论PDGFRα基因突变可能参与了GIST的发生发展，但突变是否有助于判断肿瘤的良恶性和提示预后，仍需更多的研究和观察。     

膀胱移行细胞癌中c-erbB-2、p53及p16 蛋白的表达

目的　探讨膀胱癌组织中p16、c erbB 2和p5 3蛋白表达与膀胱癌病理分级、临床分期和转移的关系。方法　应用免疫组化SP法对 75例膀胱癌组织中p16、p5 3及c erbB 2蛋白表达进行检测。结果　 75例膀胱癌中p16、p5 3及c erbB 2的阳性率分别为 41.3% (31 75 )、44 .0 % (33 75 )和40 0 % (30 75 ) ,p16和c erbB 2蛋白在膀胱癌中的阳性率与肿瘤病理分级和临床分期有显著意义 (P<0 0 5 ) ,p5 3和c erbB 2阳性率与肿瘤临床分期及转移有密切的关系 (P <0 .0 1)。 77.3% (5 8 75 )肿瘤有上述蛋白的阳性表达 ,其中 5 3.3% (40 75 )的肿瘤同时有多个蛋白的阳性表达。结论　肿瘤的多因素分析比单因素分析更有价值 ,癌基因c erbB 2和抑癌基因p5 3、p16蛋白的表达异常及协同作用在膀胱癌的发生发展中起重要作用     

鼻咽癌中p27^kip1表达及其与临床病理特征相关研究

目的检测p27^kip1蛋白在鼻咽癌中的表达,分析其与临床病理特征的关系。方法收集60例手术活检切取的鼻咽癌组织蜡块,与30例非肿瘤鼻咽组织石蜡标本作比较。应用免疫组织化学技术检测鼻咽癌组织中p27^kip1蛋白的表达,回顾性研究鼻咽癌患者的临床病理特征。结果p27^kip1蛋白在鼻咽癌细胞核和细胞质中均有表达。鼻咽癌组织中细胞核表达阳性率为46．7％（28／60）,低于非肿瘤鼻咽组织细胞核表达的90％（27／30）,P〈0．01;而在细胞质中阳性率为68．3％（41／60）,显著高于非肿瘤鼻咽组织的细胞质表达的20％（6／30）,P〈0．01。未发现p27^kip1蛋白在细胞核和细胞质表达与鼻咽癌原发灶的范围和局部侵犯程度有关,但p27^kip1蛋白胞核表达与淋巴结转移、远处转移和TNM临床分期有关;细胞质表达仅与淋巴结转移和TNM临床分期有关,可见p27^kip1蛋白胞核表达与临床病理的关系更为密切。结论与非肿瘤鼻咽组织相比,p27^kip1蛋白为细胞核低表达、细胞质高表达。鼻咽癌中p27^kip1蛋白存在不同于非肿瘤鼻咽组织的错误的核质定位,其在细胞核中表达的减少或丢失可能与鼻咽癌的恶性发展有关,提示p27^kip1蛋白的异常表达和定位可能涉及鼻咽癌的分期和转移。     

应用半套式聚合酶链反应检测非何杰金淋巴瘤bcl—2／JH融合基因

为研究我国非何杰金淋巴瘤（NHL）患者bcl－2基因重排的发生情况，以及其可能的临床应用价值，应用半套式聚合酶链反应（semi－nestedPCR）检测45例非何杰金淋巴瘤bcl-2／JH融合基因，发现12例阳性，其中15例滤泡性淋巴瘤中有9例阳性（60％）；30例弥漫性淋巴瘤中3例阳性（10％），5例反应性淋巴组织增生者阴性。并发现bcl-2基因断裂点绝大部分位于主要断裂区（11／1291．7％），少数位于次要断裂区（1／12，8．3％）。结果提示，形成bcl-2／JH融基因是B细胞非何杰金淋巴瘤发生的重要病因，检测bcl－2／JH融合基因对鉴别淋巴组织的良恶性增生、明确克隆来源，提示预后及检测微小残留病灶均有重要意义。     

bcl-10蛋白在MALT淋巴瘤中的表达

目的 观察bcl-10蛋白在黏膜相关淋巴组织样结外边缘区B细胞淋巴瘤（MALT淋巴瘤）中的表达及其意义。方 法对62例MALT淋巴瘤进行回顾性研究,采用免疫组织化学SP法检测bcl-10及Ki-67的表达。结果 62例MALT淋巴瘤中,60例（96．8％）表达bcl-10,其中胞核和胞质同时表达33例（53．2％）,仅有胞质表达27例（43．6％）。10例桥本甲状腺炎bcl-10均表达于胞质。bcl-10核阳性组发病平均年龄（51．4岁）比bcl-10核阴性组（56．6岁）小5．2岁,bcl-10核阳性组以男性占优势,bcl-10核阴性组以女性发病较多。不同解剖部位核阳性表达率差异有统计学意义（P〈0．05）,肺和胃肠道检出率较高,而甲状腺较少;不同临床分期之间核阳性表达率差异P〉0．05;bcl-10核表达与不表达两组在肿瘤细胞形态上差异P〉0．05;在胃肠道40例病例中,不同浸润组别的核阳性率表达差异P〈0．05,浸出浆膜外的病例核阳性率高于局限在浆膜以内的病例;bcl-10核阳性组与核阴性组Ki-67增殖指数的差异无统计学意义。随访52例（83．9％）,bcl-10核阳性组（29例,96．3％）与核阴性组（23例,66．4％）的5年生存率差异无统计学意义。结论 bcl-10在MALT淋巴瘤中主要有胞质和胞核两种表达模式;胞核的表达在不同解剖部位之间检出率不同,并与肿瘤的浸润深度相关。bcl-10在MALT淋巴瘤的诊断、治疗和预后方面有一定的提示作用。     

Cell death in retinoblastoma: electron microscopic, immunohistochemical, and DNA fragmentation studies

Apparent cell loss by apoptosis occurs in carcinomatous tissue. To investigate cell death in retinoblastoma (Rb), ultrastructural examination, ApopTag staining, electrophoresis to detect apoptotic DNA fragmentation, and flow cytometric studies were performed. Immunostaining for the oncogenic products bcl-2 and p53 was also carried out. Relationships between the proliferation fraction (PF), apoptotic index (AI), and the distribution of bcl-2 and p53 were investigated according to the degree of histologic differentiation of Rb. Ultrastructurally, two patterns of cell death were seen. Necrotic cells exhibited vacuolation of cytoplasmic organelles with a marked lytic change in the karyoplasm and cytoplasm. In contrast, apoptotic cells were characterized by crescentic margination of chromatin, condensation of karyoplasm and cytoplasm, and fragmentation of the nucleus. Differentiated Rb had a low AI value (< 1%), whereas undifferentiated Rb had a high AI value (> 8%). The PF of undifferentiated RB (31%) was significantly higher than that of differentiated RB (14%). Analysis of DNA fragmentation using 3'-end labeling with terminal transferase indicated that undifferentiated Rb has increased DNA cleavage. The distribution of apoptotic bodies within Rb was inversely correlated with the expression of bcl-2. A majority of tumor cells of differentiated Rb were negative for p53, whereas 20-40% of tumor cells of undifferentiated Rb showed a positive reaction for p53. These findings suggest that the degree of susceptibility to apoptosis is closely related to PF, is inversely related to the degree of differentiation of Rb, and is protected by oncogene bcl-2.     

胃肠道间质瘤的电镜和免疫组织化学研究

目的 探讨胃肠道间质瘤的组织起源和神经分化特征。方法 应用电镜和免疫组织化学对20例胃肠道间质瘤的超微结构和神经分化相关抗原表达进行研究。结果 20例胃肠道间质瘤c—kit表达均阳性。其中7例超微结构存在神经分化,12例未见神经或肌细胞分化,仅有1例存在向平滑肌纤维分化。神经分化形态表现为瘤细胞胞质和胞质突起内可见散在或簇状分布的致密核心颗粒,并形成突触样结构。并可见神经元样突起、饮液空泡和团丝样纤维。神经分化伴有致密核心颗粒病例在良性、交界性和恶性组各有1例、1例和5例。神经分化组病例神经分化相关抗原神经元特异性烯醇化菌、CD99、S—100和CD56阳性表达分别有7例、7例、5例和4例,均高于未定分化组。结论 胃肠道间质瘤和所谓的胃肠道自主神经肿瘤在组织形态和免疫表型都存在相互重叠现象,神经分化超微结构观察和神经分化相关抗原分析有助于确定胃肠道间质瘤的神经分化改变以及潜在的生物学行为。     

bcl-2和p53表达在结直肠癌预后评估中的价值

目的　探讨bcl 2和 p5 3表达水平与结直肠癌肿瘤生物学特征及其预后的关系。 方法　应用免疫组化S P法 ,检测93例结直肠癌标本中bcl 2与p5 3蛋白的表达 ,并与临床病理特征进行相关分析。结果　bcl 2蛋白在结直肠癌的阳性表达率为 5 7 0 % (5 3/ 93) ,与淋巴结转移呈负相关 (P <0 0 1) ,p5 3表达阳性率为 4 3% (4 0 / 93) ,与淋巴结转移无相关 (P >0 0 5 ) ,但p5 3表达阳性患者的预后差于 p5 3阴性组 (P =0 0 1)。当分析bcl 2和 p5 3联合表达时 ,Dukes分期及淋巴结转移各组间有差异 (P <0 0 5 )。其中bcl 2 (+) / p5 3(- )表达形式多见于DukesA和B期肿瘤 (4 1 0 % ,2 3/ 5 6 )。经Cox模型多因素分析结果表明 ,淋巴结转移 (P <0 0 1)和 p5 3表达 (P <0 0 1)是结直肠癌独立预后影响因素。其他指标 ,包括bcl 2和 p5 3联合表达情况等均未显示出统计学意义。结论　bcl 2和p5 3表达水平可提示结直肠癌的生物学行为 ,但bcl 2的表达与淋巴结转移状况相关不是一个单独作用的预后指标 ,而 p5 3虽与其它生物学特性关系不大 ,却是一个相对独立的结直肠癌预后指标。     

胃肠道间质瘤中p53和bcl-2表达与预后的关系

目的探讨细胞凋亡相关基因p53和bc l-2在胃肠道间质瘤(gastrointestinal strom al tumors,G ISTs)中的表达与预后关系。方法对194例G ISTs构建组织微阵列(TMA),采用免疫组化EnV ision法检测G ISTs组织中p53和bc l-2基因蛋白的表达。结果在p53 TMA中,184例可评估(94.8%),在bc l-2 TMA中181例可评估(93.3%)。p53和bc l-2基因蛋白阳性率分别为34.8%和59.1%。p53蛋白阳性表达率与肿瘤大小、NIH分级、肿瘤部位、坏死、细胞密集程度、核分裂象和转移复发有关。bc l-2阳性表达率与肿瘤大小、NIH分级、肿瘤部位、坏死和黏膜受累及有关。p53蛋白阳性和阴性表达者的5年生存率分别为40.1%和76.5%,两者比较差异有显著性(P<0.01)。bc l-2阳性和阴性表达者的5年生存率分别为55.1%和76.2%,两者比较差异有显著性(P<0.05)。p53蛋白阳性组和阴性组与bc l-2蛋白的阳性表达差异有显著性(P<0.01)。结论p53、bc l-2表达与G ISTs预后有关,p53、bc l-2可作为判断G ISTs预后的标志物。     

Cell Death in Retinoblastoma: Electron Microscopic,Immunohistochemical, and DNA Fragmentation Studies

Apparent cell loss by apoptosis occurs in carcinomatous tissue. To investigate cell death in retinoblastoma (Rb), ultrastructural examination, ApopTag staining, electrophoresis to detect apoptotic DNA fragmentation, and flow cytometric studies were performed. Immunostaining for the oncogenic products bcl-2 and p53 was also carried out. Relationships between the proliferation fraction (PF), apoptotic index (AI), and the distribution of bcl-2 and p53 were investigated according to the degree of histologic differentiation of Rb. Ultrastructurally,two patterns of cell death were seen. Necrotic cells exhibited vacuolation of cytoplasmic organelles with a marked lytic change in the karyoplasm and cytoplasm. In contrast, apoptotic cells were characterized by crescentic margination of chromatin, condensation of karyoplasm and cytoplasm, and fragmentation of the nucleus. Differentiated Rb had a low AI value (&lt; 1%),whereas undifferentiated Rb had a high AI value (&gt; 8%). The PF of undifferentiated RB (31%) was significantly higher than that of differentiated RB (14%). Analysis of DNA fragmentation using 3'-end labeling with terminal transferase indicated that undifferentiated Rb has increased DNA cleavage. The distribution of apoptotic bodies within Rb was inversely correlated with the expression of bcl-2. A majority of tumor cells of differentiated Rb were negative for p53, whereas 20-40% of tumor cells of undifferentiated Rb showed a positive reaction for p53. These findings suggest that the degree of susceptibility to apoptosis is closely related to PF, is inversely related to the degree of differentiation of Rb, and is protected by oncogene bcl-2.     

胃肠MALT淋巴瘤中bcl-10 mRNA和蛋白的表达

目的　探讨bcl- 10基因在胃肠黏膜相关淋巴组织(MALT)淋巴瘤中的表达情况及意义。方法　采用免疫组化S P 法及原位杂交技术检测40例胃肠MALT淋巴瘤和14例正常淋巴结中bcl- 10基因的表达。结果　40例MALT淋巴瘤中有36 例(90.0%)表达bcl- 10蛋白,其中21例仅在胞质表达,15例在胞质胞核同时表达;39例(97.5%)表达bcl 10mRNA。bcl -10 蛋白与mRNA表达之间差异无统计学意义(P>0.05)。MALT淋巴瘤临床分期与bcl- 10蛋白核表达明显相关(P<0.01)。14 例淋巴结中,8例(57.1%)表达bcl -10蛋白。淋巴滤泡内生发中心B细胞呈高度表达,边缘区B细胞中等强度表达,套区细胞 微弱表达。结论　bcl -10的高度表达在MALT淋巴瘤发生发展可能起着重要作用。bcl -10蛋白核表达与进展期MALT淋巴瘤 相关。bcl -10蛋白在淋巴滤泡各区域的表达差异提示它对B细胞分化成熟有着重要意义。     

Association of Epstein-Barr virus infection with p53 protein accumulation but not bcl-2 protein in nasopharyngeal carcinoma

AIMS: The aim of this study was to investigate the association of Epstein-Barr virus (EBV) infection with status of p53 protein expression in nasopharyngeal carcinoma (NPC). The expression of EBV gene and gene product, p53 protein and bcl-2 protein in NPC was histopathologically studied. METHODS AND RESULTS: In-situ hybridization using oligonucleotide probe to EBV-encoded small RNAs (EBERs) and immunohistochemistry using monoclonal antibodies against EBV latent membrane protein 1 (LMP1), p53 protein and bcl-2 proteins were performed in 56 primary NPCs. EBERs were detected in 46 (82%) cases and LMP1 in 17 (30%) cases. While 30 of 32 (94%) cases in differentiated nonkeratinizing carcinoma (NKC, WHO type 2) and 16 of 17 (94%) cases in undifferentiated carcinoma (UC, WHO type 3) showed EBERs expression, neither five cases of keratinizing squamous cell carcinoma (KSCC, WHO type 1) nor two cases of adenocarcinoma showed EBERs. bcl-2 protein was detected in 50 (89%) cases, but its expression did not depend on expression of LMP1. p53 protein was detected in 31 (55%) cases, and there was a correlation between expression of EBERs and p53 protein (P < 0.05) but not between LMP1 and p53 protein. CONCLUSION: In this study, close association of NKC and UC but not KSCC with the latent infection with EBV was demonstrated. The induction of bcl-2 protein by LMP1, as shown in vitro, was not demonstrated. The association between overexpression of p53 protein and the presence of EBV suggests that some EBV-encoded protein, which may be different from LMP1, may play a role for nuclear accumulation of p53 protein.     

Immunohistochemical analysis of bcl-2 and p53 expression in breast carcinomas: their correlation with Ki-67 growth fraction

We examined 59 breast cancers for p53 and bcl-2 protein expression by immunohistochemistry. The results were correlated with Ki-67 immunostaining. p53-negativity was noted in 40 cases and the remaining 19 tumours were p53-positive. Thirty-six tumours showed strong expression of bcl-2 and in 23 no staining for this protein was observed. We found statistically significant reverse correlation between expression of p53 and bcl-2 in majority of carcinomas: 31 cases were bcl-2 positive and p53-negative, and 14 tumours were bcl-2-negative and p53-positive. Six carcinomas showed no nuclear staining for Ki-67 and in the remaining 53 the percent of cancer cells positive for Ki-67 ranged from 1 to 60 (mean: 14.6). In these 53 cases we found that bcl-2-positive tumours were characterized by lower proliferation than bcl-2-negative tumours, the mean value of Ki-67 immunostaining being 10.7% and 23.0%, respectively. p53-negative tumours showed lower proliferation than p53-positive tumours: mean Ki-67 index was 10.2% and 23.9%, respectively.We conclude that immunohistochemically detected p53 and bcl-2 proteins show a significant inverse relationship in majority of breast carcinomas and their expression correlates with tumour proliferation (Ki-67 immunostaining).     

p53 expression in gastrointestinal stromal tumors

The understanding of gastrointestinal stromal tumors (GIST) is complex because of their divergent differentiation and unpredictable behavior. However, our understanding is becoming clearer, despite some cases of tumors which are exceptions from the typical cases. Tumor size, mitotic rate and, to a lesser degree, location, are the most important predictive parameters for the behavior of GIST. In this study, expression of p53 protein was evaluated in 15 cases of GIST. Tumors were divided into three groups: (i) benign (mitotic index [MI] < 5/50 high-power fields [HPF] and size < 5 cm); (ii) borderline (MI < 5/50 HPF and size > or = 5 cm); and (iii) malignant (MI > or = 5/50 HPF, irrespective of size). The mean values of p53 expression in the three groups were significantly different (benign, 10.6%; borderline, 33.8%; and malignant, 71%). The conclusion of the present study is that p53 overexpression correlates well with the malignant potential of GIST.     

人神经母细胞瘤中bcl—2基因表达与肿瘤细胞凋亡间的关系及p16…

目的探讨人神经母细胞瘤中ｂｃｌ－２基因表达状况与细胞凋亡的关系，以及ｐ１６基因的表达况状。