NSE和嗜铬素A在胃肠胰神经内分泌细胞和肿瘤中…

应用免疫组化方法，观察ＮＳＥ和嗜铬素Ａ在胃肠胰神经内分泌细胞和肿瘤组织中的表达。结果发现：ＮＳＥ在肿瘤组阳性率为９０．９％，嗜铬素Ａ为６０．６％；而在正常细胞组，ＮＳＥ阳性率３２．４％，嗜铬素Ａ为１００％，我们认为ＮＳＥ和嗜铬素Ａ对胃肠胰神经内分泌肿瘤的诊断都具有特殊的意义，联合应用能提高其诊断率。     

适用免疫组化的突触素与嗜铬素A 单克隆抗体的制备与临床初步评价

目的:制备突触素与嗜铬素A 免疫组化单克隆抗体,期望可初步应用于临床。方法:设计Syn 与CgA 融合基因,并构建至表达载体上表达获得融合蛋白。经过抗原免疫、细胞融合后筛选获得目标抗体。通过临床样本对比验证,研究Syn 和CgA 两者的特异性,评价相关系数。结果:通过筛选,共获得2 株分别针对Syn 与CgA 的抗体3D9 和4A12。通过对比抗体试剂共同验证19 种蜡块组织,统计结果分析,结果符合性较好,相关系数分别达到r =0.989 2 与r =0.993 9。结论:该方法成功制备获得了可初步适用于免疫组化的突触素与嗜铬素A 抗体,该方法可为免疫学抗体研究提供一定的借鉴意义。     

p-糖蛋白在结肠癌组织中的表达

ｐ┐糖蛋白在结肠癌组织中的表达沈金辉吴璇蔡广玲曾绍文我们应用免疫组化方法检测多药耐药（ＭＤＲ）基因产物ｐ－糖蛋白在结肠癌组织中的表达，探讨ＭＤＲ基因与结肠癌的关系，为结肠癌的防治提供客观的依据。１材料和方法选用１９９２～１９９５年间本院结肠癌根治术标...     

NSE和嗜铬素A在胃肠胰神经内分泌细胞和肿瘤中的诊断意义

应用免疫组化方法,观察NSE和嗜铬素A在胃肠胰神经内奋泌细胞和肿瘤组织中的表达。结果发现:NSE在肿瘤组阳性率为90.9％,嗜铬素A为60.6％;而在正常细胞组,NSE阳性率32.4％,嗜铬素A为100％。我们认为NSE和嗜铬索A对胃肠胰神经内分泌肿瘤的诊断都具有特殊的意义,联合应用能提高其诊断率。     

蛋白质组学检测β-原肌球蛋白在结肠癌组织中的表达

目的：通过比较结肠腺癌与正常结肠黏膜的蛋白质组表达差异,寻找与结肠腺癌发生相关的蛋白质,筛选结肠癌诊断的分子标志物。方法: 运用蛋白质组学技术,对8例结肠腺癌组织和8例正常结肠黏膜组织进行胶内差异双向电泳（2-D）,选择差异表达超过2倍的蛋白质进行MALDI-TOF/TOF质谱分析和生物信息学分析,并对结肠癌组织中表达下调的蛋白β-原肌球蛋白(TM β)进行Western blotting验证。结果: 成功建立结肠腺癌和正常结肠黏膜的双向凝胶电泳图谱,结肠腺癌和正常黏膜组织凝胶电泳图谱中平均蛋白质斑点数分别为3 289和3 066,其中表达差异超过2倍的斑点共有31个,质谱分析和数据库检索共鉴定出18种蛋白质,包括cytokeratin 8、cytokeratin 10、S100A6、TM β、protein disulfide isomerase 等。从功能分析,这些差异蛋白与癌细胞的发生、增殖、分化、转移等相关;TM β在结肠癌组织中的表达水平Western blotting结果与电泳结果一致。结论: 蛋白质组学能有效地分离和鉴定结肠腺癌组织与正常结肠组织间的差异表达蛋白质,结肠癌组织中表达下降的TMβ,可能成为结肠癌诊断的分子标记物。     

血管内皮生长因子受体3和Podoplanin在人结肠癌组织中的表达

目的:观察血管内皮生长因子受体3(vascular endothelial growth factor receptor-3,VEGFR-3)和Podoplanin在人结肠癌组织中的表达,以探讨二者标记淋巴管的特异性。方法:选择55例人结肠癌组织标本,应用免疫组织化学法观察VEGFR-3和Podoplanin在人结肠癌组织中的表达。结果:Podoplanin主要表达于淋巴管内皮细胞上,微血管极少着色;VEGFR-3主要表达于淋巴管内皮细胞上,另外在小血管内皮也有较丰富的表达。结论:Podoplanin在淋巴管内皮细胞的表达具有较高特异性,可以用来标记淋巴管。     

结肠癌组织中Fas表达的定量分析

目的：探讨Fas在结肠癌中的表达及其与肿瘤发生、发展及肿瘤分化程度的关系。方法：用直接免疫荧光－流式细胞技术对50例结肠腺癌组织及手术切缘的正常结肠黏膜组织中Fas的表达进行定量检测，分析Fas表达与肿瘤大体类型、浸润深度、分化程度及淋巴结转移的关系。结果：Fas在结肠癌组织中的平均阳性率及平均荧光指数均低于其手术切缘的正常结肠黏膜组织，在12例伴有转移的结肠癌组织中Fas的平均阳性率及平均荧光指数均低于38例没有转移的结肠癌；9例肿瘤限于肌层以内的结肠癌组织中Fas平均阳性率及平均荧光指数均高于41例肿瘤侵及肌层以外的结肠癌病例。不同分化程度、不同大体类型的结肠癌组织中Fas的平均阳性率及平均荧光指数差异无显著性。结论：结肠腺癌组织中Fas的表达受到了抑制，Fas的表达与肿瘤的转移及浸润深度呈负相关，与肿瘤的分化程度及大体类型无关。     

结肠嗜酸细胞癌2例临床病理分析

目的 探讨结肠嗜酸细胞癌的病理组织学特点、诊断及鉴别诊断。方法 对2例结肠嗜酸细胞癌手术切除标本进行光镜、免疫组化染色及电镜观察。结果 结肠嗜酸细胞癌组织学特点为细胞圆形、多边形,胞质丰富,胞质内见弥漫分布的强嗜酸性颗粒,细胞核中度异型性,核仁明显;癌细胞呈巢状排列,伴有腺管及乳头形成。免疫表型：癌细胞CK19、EMA阳性表达,不表达SMA、S-100蛋白及CgA。PAS染色阴性。电镜观察细胞质充满大量线粒体,缺乏内分泌及外分泌颗粒。结论 嗜酸细胞癌是不同于低分化腺癌及神经内分泌癌的一种特殊类型细胞癌。     

性激素受体在前列腺病变组织中的表达

用抗雄、雌及孕激素受体（ＡＲ、ＥＲ和ＰＲ）的多克隆抗体，采用ＡＢＣ法检测３０例良性前列腺增生（ＢＰＨ）和３０例前列腺癌（ＰＣ）石蜡标本中性激素受体（ＳＲ）的表达。结果显示：ＢＰＨ组和ＰＣ组ＡＲ、ＥＲ和ＰＲ表达的阳性率差异均无显著性，因而不能根据ＡＲ、ＥＲ和ＰＲ阳性率区分ＢＰＨ和ＰＣ。另外，ＡＲ、ＥＲ或ＰＲ阳性者的存活时间均长于ＳＲ阴性者，提示ＳＲ表达与ＰＣ患者内分泌治疗的预后有一定关系，受体阳性率     

5氟尿嘧啶上调干细胞标记物CD133在结肠癌细胞中的表达

目的: 研究通过5氟尿嘧啶(5-FU)对结肠癌干细胞标记物CD133表达的影响,探讨5-FU对结肠癌干细胞的影响。方法: 用流式细胞仪检测CD133在结肠癌细胞株表面的表达, 磁珠细胞分离的方法分离结肠癌细胞株DLD1中CD133阳性和阴性的细胞群,细胞克隆形成实验检测2群细胞的自我更新能力,新型四唑氮盐方法(MTS)检测2群细胞对5-FU敏感性的差异,qPCR方法检测5-FU处理结肠癌细胞后CD133mRNA水平的变化。结果: 结肠癌细胞株DLD1、HT29、SW480、HCT116、Lovo、RKO细胞表面CD133的表达率分别为30.20%、82.00%、0.34%、91.80%、85.30%、0.28%。DLD1细胞中以CD133为标记有2群明显的细胞,MACS方法分离后阳性细胞群中CD133为87.21%±5.33%, 而阴性细胞群中阴性细胞的比例为84.30%±4.65%;CD133阳性的细胞与未分离及CD133阴性细胞相比,克隆形成能力强(46.33%±4.44% vs 31.00%±2.00%,P<0.05),对5-FU的敏感性下降20%,P<0.01。在DLD1和HT29细胞中,5-FU 1 mg/L上调CD133mRNA水平的表达,从1升为1.684±0.012(P<0.01)、HT29细胞从30.702±0.284升为49.379±0.460(P<0.01)。结论: 与CD133阴性细胞相比CD133阳性细胞克隆形成能力强,对5-FU的敏感性下降;5-FU上调干细胞标记物CD133mRNA水平的表达,CD133阳性的结肠癌干细胞在5-FU的治疗过程中被富集。     

结肠肿瘤中wisp-1的表达及意义

目的探讨wisp-1在原发性结肠癌组织中的表达及临床意义。方法应用免疫组化EnVision两步法检测wisp-1蛋白在138例结肠癌组织(48例含癌旁组织)、37例结肠腺瘤组织及66例结肠正常黏膜组织中的表达;应用实时荧光定量PCR检测wisp-1基因在37例新鲜原发性结肠肿瘤标本及对照组37例正常结肠中的表达;分析wisp-1蛋白、基因表达与患者临床病理特征的关系。结果 wisp-1蛋白在正常结肠、结肠腺瘤、癌旁组织及结肠癌中阳性率分别为53.0%、67.6%、75.0%、76.1%,其在结肠癌及癌旁组织中表达高于正常结肠(P<0.05);wisp-1蛋白表达与淋巴结转移及浸润深度有关(P<0.05);wisp-1基因在结肠肿瘤中平均表达是正常结肠中的1.349倍(P>0.05)。结论 wisp-1蛋白异常可能在结肠癌的发生过程中起重要作用,而其基因在结肠癌中的作用尚需进一步研究。     

Expression of Lewisx sugar structure in the liver metastasis of mouse colon carcinoma (colon 26) cells

Immunohistochemical aspects of the process of experimentally induced metastasis were examined by light and electron microscopy employing a series of labeled carbohydrate-specific monoclonal antibodies as probes. Liver metastasis was induced by injecting mouse colon carcinoma cell (colon 26) into the spleen of Balb/c mice. Labeled anti-Lewis^x (Le^x) antibody stained the metastasized colon 26 cells strongly compared with the heterogeneous and faint staining in non-metastasized tumor foci in the spleen or in the subcutaneous space. Other antibodies having specificities for Lewis-related antigens other than Le^x, e.g. those against Le^y, Le^a, Le^b, sialyl Le^x and sialyl Le^a, did not show any differences in binding between metastasized cells and non-metastasized tumor foci. Immunoelectron microscopy revealed the expression of Le^x-carbohydrate in the plasma membranes as well as in the intercellular spaces of metastasized colon 26 cells in the liver. Based on these results, it is likely that sugar chains containing the Le^x-carbohydrate structure are involved in the interactions between colon 26 cells and hepatic cells during the process of liver metastasis.     

结直肠癌合并糖尿病患者组织中EMT相关蛋白的表达

目的观察结直肠癌合并糖尿病患者组织中促进上皮间质转换(EMT)进展的相关蛋白的表达。方法收集直肠癌及合并糖尿病患者的肿瘤组织及非肿瘤组织,分为对照组和肿瘤组;根据是否合并糖尿病,又分为伴及不伴糖尿病组。用免疫组化检测晚期糖基化终末产物(AGE)及其相应受体(RAGE)、上皮标志物钙黏连蛋白E(E-cad)、β-连环蛋白(β-catenin)、波形蛋白(vimentin);蛋白质印迹法检测各组中不同分化程度下相关蛋白的表达。结果 RAGE、AGE、E-cad和vimentin主要染色于细胞膜上,β-catenin主要染色于胞质内,且两个肿瘤组染色较对照组更深(P0.01);与对照组相比,不伴糖尿病组及伴糖尿病中各蛋白表达均增高,E-cad表达降低(P0.01);在同一分化状态下,伴糖尿病组中各蛋白表达水平较不伴糖尿病组高,E-cad较之降低(P0.01)。结论结直肠癌合并糖尿病患者组织中相关蛋白β-catenin和vimentin表达上调,E-cad表达下降,从而促进EMT进展,对结直肠癌发生发展起重要促进作用。     

Expression of the nuclear oncogene p53 in colon tumours

The nuclear tumour antigen p53 is expressed by a gene localized on the p-arm of human chromosome 17, a region frequently deleted in colon carcinomas. Using a monoclonal antibody to p53 antigen, immunohistochemical analysis of carcinomas and dysplastic tubular adenomas of the colon has been performed to study the relation between p53 expression and dysplasia or malignancy. With this methods p53 was detectable in 55 per cent of colon carcinomas (n = 29). In 8 per cent of adenomas (n = 74), focal nuclear p53 expression was found in dysplastic epithelial cells. In general, these p53-positive regions of the polyps were histologically indistinguishable from the neighbouring tubuli. Sometimes the p53-positive nuclei were found in a focus of more highly dysplastic epithelium. The results suggest that expression of the p53 gene may be part of the process of malignant transformation of dysplastic colon polyps.     

Immunohistochemical detection of the receptor for urokinase plasminogen activator in human colon cancer

Paraffin-wax embedded specimens from 30 cases of colonic adenocarcinoma were investigated for immunoreactivity for the receptor of urokinase-type plasminogen activator (uPAR). In all cases there was a strong signal, predominantly at the invasive foci. The positive cells were mainly tumour-infiltrating macrophages but neutrophils and eosinophils were also strongly stained. The neoplastic cells were positive in 19 of the samples with staining of occasional or a moderate number of cells. In uninvolved, normal-appearing mucosa adjacent to the malignant infiltrates, immunostaining of both macrophages and neutrophils was seen, but the labelling was less intense than that seen in the malignant lesions. Weak to moderate staining of normal intestinal epithelium was also seen at the luminal surface. Comparison between immunoreactivity and in situ hybridization showed a similar distribution of protein and mRNA with two exceptions: first, neutrophils (strongly immunoreactive for uPAR) were negative or only weakly positive for uPAR mRNA; and second, many cancer cells at invasive foci showed prominent hybridization signals but no detectable uPAR immunoreactivity. Together with previous findings of urokinase plasminogen activator (uPA) protein and mRNA being expressed in tumour-infiltrating fibroblast-Iike cells at the invasive foci, these results support the view that the uPA pathway of plasminogen activation is involved in tissue degradation in colon cancer. The results also extend and consolidate an emerging picture of non-neoplastic tumour stromal cells producing molecules involved in the generation and regulation of extracellular proteolysis in cancer.     

结肠癌组织中LKB1的表达及其与上皮-间质转化的关系

目的探讨LKB1在结肠癌组织中的表达及其与肿瘤上皮-间质转化的关系。方法采用免疫组化法检测75例结肠癌组织及对应癌旁正常结肠组织中LKB1、上皮-间质转化(epithelial mesenchymal transformation,EMT)相关标志物E-cadherin、N-cadherin、vimentin的表达,分析LKB1表达与结肠癌临床病理特征及EMT的关系。结果 49例结肠癌组织中LKB1低表达,55例正常结肠组织中LKB1高表达;LKB1表达降低与淋巴结转移、TNM肿瘤分期相关(P0.05);E-cadherin、N-cadherin、vimentin分别在25例、48例、26例结肠癌组织中高表达,LKB1表达与E-cadherin表达呈正相关(r=0.648,P0.05),与Ncadherin(r=-0.891,P0.05)和vimentin(r=-0.660,P0.05)表达呈负相关。结论 LKB1在结肠癌组织中表达下调,与EMT的发生呈正相关,可作为结肠癌转移发生的预测指标。     

Monoclonal antibodies to cytoskeletal proteins: An immunohistochemical investigation of human colon cancer

Monoclonal antibodies raised to a number of microfilament-associated proteins were shown to recognize the appropriate proteins in extracts from human colon tissue. They were then used in an immunohistochemical study of normal colonic mucosa, adenomas, and adenocarcinomas. A strong reaction was seen in stromal cells within the tumours (both adenomas and adenocarcinomas) when frozen sections were stained with antibodies to filamin and caldesmon. In addition, a similar reaction was seen in the adenocarcinomas when stained with antibodies to talin and gelsolin. We believe that immunohistochemical staining with these antibodies reveals a tumour-induced process in the surrounding cells, possibly related to a host response to tumours.     

Osteopontin and colon cancer progression

Human colon cancer affects nearly 150,000 patients and results in 60,000 deaths in the United States per year. Despite significant advances in the management of the colon cancer patient, little change in survival rates has been appreciated over the past 50 years. The primary cause of death relates to the development of distant metastases to organs such as the liver and lungs. Colon cancer represents an important disease to study in order to better understand tumor progression and metastasis primarily because there is almost a stepwise advancement of the disease that is marked by measurable genetic and associated phenotypic alterations. Metastasis appears to be the end product of the development of ‘Herculean’ cell clones capable of independent growth, invasion, adhesion, avoidance of apoptosis, and angiogenesis. Although significant progress has been made in understanding the sequential genetic events leading to the development of cancer, the precise genes and the associated molecular pathways underlying the development of metastatic potential are still poorly understood. Moreover, our enhanced genetic knowledge has had relatively little trickle down effect on our clinical management of this deadly disease. For this reason, we undertook a comprehensive study to develop a molecular encyclopedia of new tumor markers and markers of tumor progression, some of which will hopefully prove useful in the clinical management of colon cancer patients by means of their capacity to detect and predict the stage and disease burden. This review will focus on the application of gene expression profiling technology to the problem of identifying new tumor markers and progression markers, and the discovery of osteopontin as the leading candidate clinical marker derived from a screen of approximately 12,000 named genes.     

IL—6转基因表达诱导大肠癌细胞凋亡

通过分子克隆技术将ＩＬ－６基因导入大肠癌细胞ＨＴ－２９，建立了目的基因转导株ＨＴ－２９ＩＬ－６细胞，该转导株接种子裸鼠皮下后，与非转导株相比，长出肿瘤的时间延后，最终瘤径明显较小，而且移植瘤标本镜下可见大量的核固缩，核染色质加深，核断裂等凋亡细胞，用携带ＩＬ－６基因的重组逆转病毒液感染大肠癌细胞Ｌｏｖｏ后，仅以１．２５μｍｏｌ／Ｌ的ＶＵＭＯＮ－２６（简称ＶＭ－２６）即可诱导该转染株发生明显凋亡，而