胎肺及肺癌花生凝集素受体的观察

应用生物素化花生凝集素以胎肺及４５例肺癌进行组织化学研究。结果显示：胎肺内支气管上皮顶部胞膜及胞质呈ＰＮＡ阳性。肺鳞癌及肺腺癌ＰＮＡ受体增多，且分布失去极性。本文提出ＰＮＡ受体改变可作为内胚层肿瘤的标志之一，尤其对腺癌有相对特异性。ＰＮＡ受体有胚胎及肺癌均表达可能提示ＰＮＡ是一种癌胚抗原。     

肺腺癌表型标志、生物学功能分型及临床分期和预后的关系

为了研究和探讨与肺癌发生、发展相关的表型抗原变型规律,预测肺腺癌发育与分化程度,确切反映癌组织本身恶性生物学行为,本文应用6种单克隆抗体(McAbs)和花生凝集素(PNA),对60例肺腺癌进行了免疫组化表型标志。观察他们在肺腺癌中的表达特点,并据此进行生物学功能分型,探讨与临床分期和预后的关系。     

神经内分泌细胞在胎肺的发育与在肺癌的胚性重演

<正> 目的:肺内存在肺神经内分泌细胞(Pulmonary neruroen-docrine cell,PNEC).PNEC在胎肺发育过程中异常活跃,与其在正常成体肺细胞的表现存在极大差异.本实验观察胚胎发育过程PNEC的形态、数量、种类及功能,旨在探讨PNEC对胎肺发育的作用.在肺癌,类癌、小细胞癌组织中存在的PNEC,与胎肺发育时期PNEC的分化具有较多相似处;非小细胞肺癌中,也伴有PNEC分化现象.本实验将研究PNEC在肺癌组织的“胚性重演”,探讨其生物学意义.方法:胚胎12-36周正常人胎肺组织30例,肺类癌4例、小细胞癌16例、不同分化程度鳞癌20例、腺癌24例及大细胞癌6例,采用组织化学Grimelius嗜银反应、免疫组织化学及透射电镜技术,检测分泌胃泌素释放肽(Gastrin-releasing peptide,GRP)、降钙素(Calcitonin,CT)及嗜铬素A(Chromogranin A,CGA)PNEC的分布、数量及超微结构,经显微图象分析仪,对其在胚胎时期细胞数量的变化进行定量分析.结果:1.胎12周胎肺内可见PNEC,分泌GRP;胎16周,出现分     

不同组织学类型人肺癌细胞系的维甲酸受体表达

目的：观察来源于不同组织学类型的人肺癌细胞系（肺巨细胞癌细胞系PLA－801，肺鳞状上皮细胞癌细胞系C－57，个旧肺腺癌细胞系GLC－82和肺腺癌细胞系PC－84045）的维甲酸受体的含量和分布特征。方法：用ELISA和免疫组化显色计算机图像分析方法，对4株具有不同侵袭潜能、不同组织学类型的肺癌细胞系的维甲酸受体（RARs、RARβ和RXRs)进行检测。结果：4株细胞系的RARβ表达极少，甚至缺如；RARs主要分布在癌细胞质中，但RXRs主要位于细胞核内，表达水平较高；RARs和RXRs在PLA－801细胞中表达最低。结论：肺癌中普遍存在着RARβ的缺失与RARs的表达分布异常，这些改变可能与肺癌的异常分化有关，也可能是肺癌细胞对维甲酸诱导分化不敏感的重要原因。RXRs较高水平的表达主要位于核内，提示用9－顺式维甲酸来诱导肺癌的分化效果可能会较好。     

肺癌(NSCLC)性激素受体和单抗Ki67的检测

我们对非小细胞肺癌的32例支气管镜活检标本和52例手术标本进行性激素受体和单抗Ki67检测,以探索在临床开展肺癌内分泌治疗的意义。结果表明:男性ER阳性率为17.5%女性为58%提示女性肺腺癌比男性肺癌更多地受到雌激素影响。TR阳性率男性为35%女性30%,其中女性患者年龄大多超过50岁提示女性肺癌     

12种凝集素受体在子宫内膜癌中的定位研究

应用12种生物素化的凝集素对85例子宫内膜腺癌、8例正常宫内膜和2例胚胎宫内膜进行免疫组织化学研究。结果发现:①正常宫内膜的凝集素受体分布于腺体的腔缘,腺癌宫内膜的凝集素受体主要分布于细胞浆内,且比正常宫内膜有明显增加;②Con A、RCA、WGA在胚胎、正常及腺癌宫内膜中均呈阳性反应,可能与子宫内膜的组织发生源相关;③BSL、SBA、PHA、PNA受体与宫内膜腺癌的分化程度相关。④LCA、PSA受体丢失与宫内膜腺癌的肌层浸润有关;LCA、PSA、PHA受体缺损与宫内膜腺癌的淋巴结转移有关。这些结果对宫内膜腺癌的糖连接物研究和治疗可能有一定的指导意义。     

非小细胞肺癌分子标志物EML4-ALK的研究进展

近几年来,中国肺癌的发病率和病死率均呈显著升高趋势.据世界卫生组织(WHO)统计,2008年,我国肺癌总病死率是28.7/10万,约45万余人,占全部癌症死亡人数的23.1％.西方国家,肺癌也是癌症死亡的主要原因;其中,非小细胞肺癌(NSCLC)约占75％～ 80％,5年总生存率约为10％～15％[1].目前已经发现一些分子靶向药物能明显改善治疗效果.表皮生长因子受体( EGFR)基因突变是原发性肺腺癌的发生原因之一,靶向针对EGFR的酪氨酸激酶抑制剂能在某种程度上有效控制肺腺癌疾病的发展[2].分子靶向治疗发展中最关键的步骤就是要明确肺癌的关键致癌基因.Soda等[3]发现另外一种有增殖活性的酪氨酸激酶融合基因棘皮动物微管相关蛋白类似因子4( EML4)-间变性淋巴瘤激酶(ALK)与NSCLC有关.EML4-ALK融合基因阳性的NSCLC患者通常没有EGFR突变及K-ras突变,多发生于不吸烟或轻度吸烟的年轻肺腺癌患者,预后较好,特别是用ALK抑制剂治疗可获约80％以上的疗效[4],对培美曲塞加顺铂化疗亦有明显效果[5].这些均提示EML4-ALK融合蛋白是肺腺癌特异性较高的分子标志物.     

国际肺癌研究协会/美国胸科学会/欧洲呼吸学会国际多学科肺腺癌分类(2011年版)解读

世界上发病率及病死率最高的恶性肿瘤是肺癌,腺癌是其中最常见的组织学类型,约占肺癌的半数.近年来,由于临床和基础研究的进展,特别是确定了肺腺癌中存在表皮生长因子受体(EGFR)突变及对EGFR酪氨酸激酶抑制性药物有着治疗反应,目前医药界投入了大量的人力和物力,从事临床治疗和分子生物学研究.而组织学分类、诊断术语及诊断标准的统一,是观察疗效和比较各试验是否有效的重要基础,因此迫切需要修订世界卫生组织( WHO)2004年版分类.     

Sonic hedgehog信号通路在肺发育及肺癌发生中的意义

在胎肺的发育过程中,支气管上皮细胞和间质之间的通讯联系对胎肺的正常形态构筑以及功能成熟都起到了重要的调控作用。Sonic hedgehog信号通路(Shh信号通路)则在这种联系中发挥着重要作用,其与其他细胞因子相互协调,形成了上皮与间质间调节作用的网络,确保了胎肺的正常发生和发育。近年来,在一些肺癌组织中也发现了Shh信号通路的异常激活,提示Shh信号通路在肺癌的发生发展过程中可能也起到了一定的作用。     

人鼻咽粘膜上皮及鼻咽癌PNA受体的观察

采用辣根过氧化物酶标记的花生凝集素(PNA),对68例人鼻咽粘膜上皮和鼻咽癌的石蜡切片进行染色。结果表明,鼻咽癌细胞中PNA受体的分布及含量与鼻咽粘膜上皮相比显著不同,提示鼻咽癌细胞具有不完整的糖蛋白合成。     

原发性肺癌的凝集素定位研究

用６种凝集素对１００例原发性肺癌进行了定位研究，结果发现：６种凝集在肺癌中表现出不同的阳性率，且分布与组织学类型有关。西蛎单叶豆（ＢＳＬ）、花生素（ＰＮＡ）、荆豆素（ＵＥＳ１）的质膜阳性率及ＰＮＡ的腔膜阳性率分别为鳞癌及腺癌的组织学分级的关系有显著性差异（Ｐ＜０．０５）。ＢＳＬ质膜定位及阴性者有生存５年以上及半年内死亡的两组鳞癌之间有显著性差异（Ｐ＜０．０５）。提示ＰＮＡ“ＢＳＬ、ＰＮＡ对肺癌有相     

肺癌组织中人乳头瘤病毒16和18型DNA相关序列的检测

采用聚合酶链反应技术（ＰＣＲ）及用ＰＣＲ法制备生物素标记探针的斑点杂交法,检测５０例肺癌、１８例肺良性病及４例胎肺组织中高危险型人乳头瘤病毒（ＨＰＶ）１６、１８型ＤＮＡ相关序列,以探讨ＨＰＶ与肺癌之间发生的关系。结果３２％的肺癌组织中检测出ＨＰＶ１６、１８ＤＮＡ,其中ＨＰＶ１６ＤＮＡ阳性１０例,ＨＰＶ１８ＤＮＡ阳性５例,１例同时含ＨＰＶ１６、１８ＤＮＡ。２７例鳞癌组织中ＨＰＶＤＮＡ阳性率为４８．２％。而肺良性病组织及胎肺组织均未发现ＨＰＶＤＮＡ。ＨＰＶＤＮＡ呈阳性的肺癌患者绝大多数为重度吸烟者。提示原发性肺癌的发生可能与ＨＰＶ感染有关。     

花生凝集素在胃癌及其癌旁粘膜标记的定位研究

The histochemical binding of fluorescein isothiocyanate conjugated peanut agglutinin (PNA) to paraffin sections of 52 cases of gastric carcinomas and their adjacent mucosa was investigated. PNA could recognize the terminal Gal and beta-D-Gal (1-3)-GalNac. A high positive rate was obtained in PNA binding to malignant cells particularly in cases of signet ring cell carcinoma and mucinous adenocarcinoma. Three characteristic patterns of PNA binding to the gastric carcinoma were identified. Type I: intracytoplasmic, covering the entire cytoplasm of the malignant cells. Type II: linear, distribution along the luminal surface of the malignant cells. Type III: only very weak reaction present in a few cells or negative. Significance of PNA binding patterns in respect of the histologic classification and differentiation degree of the gastric carcinoma are discussed. Neuraminidase digestion was noticed to enhance PNA binding only in well-differentiated adenocarcinoma and PNA receptor have no specificity for tumor.     

Differences in lectin binding of malignant pleural mesothelioma and adenocarcinoma of the lung.

In order to differentiate between malignant pleural mesothelioma and adenocarcinoma of the lung, the glycoconjugate profiles of 6 reactive mesothelial lesions, 23 mesotheliomas (17 epithelial, 1 desmoplastic, 2 biphasic, and 3 fibrous types), and 28 well-differentiated pulmonary adenocarcinomas were evaluated with the use of 8 lectins in addition to anti-carcinoembryonic, anti-keratin and anti-epithelial membrane antigen. Formalin-fixed, paraffin-embedded tissues were stained with the avidin-biotin peroxidase complex method. Reactions of wheat germ (WGA) and peanut (PNA) agglutinin with neuraminidase treatment lectins were positive in 5 of 6 (83%) and 3 of 6 (50%) cases, respectively, in reactive mesothelial lesions. Thirteen of 23 (57%) malignant mesotheliomas of the pleura showed a positive reaction for WGA and PNA with neuraminidase treatment; other lectins were low-positive, below 9%. In contrast, pulmonary adenocarcinomas showed positive reactions in 27 of 28 cases (96%) for PNA, 26 of 28 (93%) for Ricinus communis (RCA-I), 25 of 28 (89%) for WGA, and 22 of 28 (79%) for succinylated WGA (SucWGA). The findings suggest that malignant pleural mesothelioma and pulmonary adenocarcinoma have consistent and distinct glycoconjugate profiles, and that stains for RCA-I and SucWGA may be useful for differential diagnosis.     

Image analysis for quantitation of estrogen receptor in formalin-fixed paraffin-embedded sections of breast carcinoma.

Monoclonal antibody to human estrogen receptor (ER) provides a useful immunohistochemical tool for the evaluation of ER content in breast carcinoma, but visual interpretation is subjective. Computer-assisted image analysis has proved effective in immunohistochemical quantitation of ER in fresh tumor imprints and cryostat sections. We examined the usefulness of this technique in 5-microns-thick formalin-fixed paraffin-embedded tissue sections of 66 cases of primary breast carcinoma previously assayed by dextran-coated charcoal (DCC) analysis. Immunohistochemistry was automated and performed on a Code-on slide stainer (Instrumentation Laboratories, Lexington, MA) using Pronase predigestion, a monoclonal antibody (ER-ICA; Abbott, Chicago, IL), and a biotin-labeled secondary antibody. Detection was achieved with an avidin-alkaline phosphatase conjugate and nitroblue tetrazolium (NBT) bromochloroindoyl phosphate (BCIP) substrate. The immunohistochemical ER staining was analyzed visually and with the CAS/200 image analyzer (Elmhurst, IL). The visual semiquantitative histologic scores (HSCORE), the automated quantitative assays including the percentage of positive nuclear areas (PNA), and the quantitative immunocytochemical scores (QIC SCORE = PNA x % of positive stain/10) were compared with the corresponding DCC results. Linear correlations were demonstrated between all immunohistochemical assays and the logarithm of DCC, the strongest correlation seen with PNA (r = 0.91). Threshold points for positive HSCORE, QIC SCORE, and PNA assays were extrapolated using DCC as the reference. ER immunodetection by PNA as compared with visual examination alone was enhanced by 18% (up to 88%) in sensitivity and 34% (up to 94%) in specificity, and the DCC concordance rate increased by 26% (up to 91%). A comparative chart extrapolating DCC from PNA was thus established.(ABSTRACT TRUNCATED AT 250 WORDS)     

PNA Lectin-Based Separation of Thymocytes into Mature and Immature Subpopulations: CD4-8- Double Negative Cells Display Characteristics of PNAlo Mature Thymocytes

Cortical (immature) thymocytes are widely reported to express intermediate to high levels of receptors for the lectin, peanut agglutinin (PNA). Light-scatter studies of murine fetal thymocytes stained with PNA or anti-mouse CD4 and CD8 monoclonal antibodies indicated, however, that the most immature CD4-8- (DN) thymocyte subpopulation binds levels of PNA commonly described as PNA^lo. Evaluation of the PNA binding characteristics of fetal thymocytes negative for the CD8 antigen confirmed the existence of a major population (approximately 20% of total cells) of CD4^-8^- PNA^lo fetal thymocytes. The majority of these DN thymocytes were subsequently found to bind sub-agglutinating levels of PNA, similar to mature CD4⁺ or CD8⁺ single positive (SP) thymocytes. Given this information, an immunomodulating compound (2,3,7,8-tetrachlorodibenzo-p-dioxin; TCDD) known to produce a maturational delay in murine thymocytes was tested for a possible concurrent effect on thymocyte PNA lectin binding. A TCDD- induced increase in DN thymocytes was found to be paralleled by an increase of equal magnitude in PNA^lo thymocytes. Taken together, these data provide evidence that acquisition of the PNA receptor may be a maturational event occurring during the DN stage of thymocyte ontogeny. Further, these results suggest that separation of thymocytes into mature (medullary) and immature (cortical) subpopulations by PNA agglutination may result in contamination of medullary cells by the most immature (DN) subpopulation of thymocytes.     

Immunohistochemical progesterone receptor assay. Measurement by image analysis.

To determine the efficiency of image analysis in immunohistochemical progesterone receptor (PgR) measurement, 94 primary breast carcinoma tissue samples were evaluated for PgR by biochemical dextran-coated charcoal assay (DCC) and an immunohistochemical method. Frozen sections immunostained for PgR with a monoclonal antibody (Abbott PgR-ICA, Chicago, IL) and the peroxidase-antiperoxidase technique were scored semiquantitatively histologic score by microscopy and quantitatively (percentage nuclear area immunopositivity [PNA] using the CAS 200 image analyzer (Cell Analysis Systems, Elmhurst, IL). There was a positive correlation between dextran-coated charcoal assay and both histologic score (r = 0.82) and PNA (r = 0.69). Selected cutoff points of 60 histologic score and 6.5% PNA based on sensitivity/specificity calculations yielded a predictive value of a negative test of 73% and 80%, respectively, and a positive predictive value of 100% for both; ranges of fmol/mg protein PgR correspond to ranges of histologic score and PNA. The use of an image analyzer to measure PNA in PgR-immunostained sections is a viable alternative to dextran-coated charcoal assay, especially when insufficient fresh tissue is available.     

Immunoexpression of the α subunit of a guanine nucleotide-binding protein (Go) in pulmonary neuroendocrine cells and neoplasms

The α subunit of a GTP-blndlng protein, Go, was investigated in pulmonary neuroendocrine neoplasms and fetal tissues of the lung by an immunohistochemlcal method. Positive immunostaining for the α subunit of Go (Goα) was found predominantly on the cell membrane and found occasionally in the cytoplasm. Typical carcinoids were all positively stained (9/9), and small cell carcinoma showed weaker and less frequent staining (5 positive cases in 10). Atypical carcinoids were variously stained (3/4). The tendency for obvious neuroendocrine differentiation to be immunohistochemically determined in typical carcinoids and not in small cell carcinoma is also true of staining for neuron specific enolase (NSE), chromogranin A (CG-A) and synaptophysin. In the lung, Goα-immunostaining was positive not only in nerve tissues but also in the airway epithelium. In the fetal lung, serial sections immunostained for NSE, CG-A and Goα confirmed that Goα-immunoreactive cells belong to the neuroendocrine cell population. The biological significance of Goα is unclear in normal and neoplastic lung tissues, but Goα is a useful marker of neuroendocrine cells and neoplasms of the lung.     

Lectin histochemistry of normal lung and pulmonary adenocarcinoma

In order to evaluate the staining pattern of glycoconjugate profiles in adenocarcinomas of the lung, pulmonary adenocarcinomas were classified according to their: (a) degree of differentiation; (b) cellular subtyping and mucus secretion; and (c) immunohistochemical characteristics. Studies were performed on 42 pulmonary adenocarcinomas using eight lectins. Formalin-fixed, paraffin-embedded tissues were stained with avidin-biotin peroxidase complex methods. Four lectins [wheat germ (WGA), succinylated WGA (SucWGA), peanut (PNA) with neuraminidase (N) treatment, and Ricinus communis (RCA-I)] showed strong positive staining reactions in well-differentiated adenocarcinomas. Bronchial surface epithelial type, one of the subtypes among 26 cases of well- and moderately differentiated adenocarcinomas, displayed strong positive staining for WGA, SucWGA, PNA N(+), RCA-I, and Bandeirea simplicifolia (BSA-I). Goblet cell types stained positive for all lectins except Dolichos biflorus (DBA). Bronchial gland cell types also showed a strongly positive stain for WGA, SucWGA, soybean (SBA), PNA N(+), RCA-I, and Ulex europaeus (UEA-I). The lectin positive staining reaction was related to the degree of mucus secretion within the tumor cells. These results revealed that the glycoconjugate profile of pulmonary adenocarcinomas was basically sialic acid, together with N-acetyl-glucosamine and beta-D-galactose. The observation that UEA-I showed a strong staining reaction in mucus-producing adenocarcinomas, such as goblet cell and bronchial gland cell types, indicates that localization of alpha-L-fucose may be a specific carbohydrate from non-mucus-producing pulmonary adenocarcinomas.     

PNA Lectin-Based Separation of Thymocytes into Mature and Immature Subpopulations: CD4–8- Double Negative Cells Display Characteristics of PNAlo Mature Thymocytes

Cortical (immature) thymocytes are widely reported to express intermediate to high levels of receptors for the lectin, peanut agglutinin (PNA). Light-scatter studies of murine fetal thymocytes stained with PNA or anti-mouse CD4 and CD8 monoclonal antibodies indicated, however, that the most immature CD4–8- (DN) thymocyte subpopulation binds levels of PNA commonly described as PNA^lo. Evaluation of the PNA binding characteristics of fetal thymocytes negative for the CD8 antigen confirmed the existence of a major population (approximately 20% of total cells) of CD4^?8^? PNA^lo fetal thymocytes. The majority of these DN thymocytes were subsequently found to bind sub-agglutinating levels of PNA, similar to mature CD4⁺ or CD8⁺ single positive (SP) thymocytes. Given this information, an immunomodulating compound (2,3,7,8-tetrachlorodibenzo-p-dioxin; TCDD) known to produce a maturational delay in murine thymocytes was tested for a possible concurrent effect on thymocyte PNA lectin binding. A TCDD- induced increase in DN thymocytes was found to be paralleled by an increase of equal magnitude in PNA^lo thymocytes. Taken together, these data provide evidence that acquisition of the PNA receptor may be a maturational event occurring during the DN stage of thymocyte ontogeny. Further, these results suggest that separation of thymocytes into mature (medullary) and immature (cortical) subpopulations by PNA agglutination may result in contamination of medullary cells by the most immature (DN) subpopulation of thymocytes.