抗p21ras单克隆抗体KGH-R1在乳腺癌组织中的免疫反应性

目的 研究p21ras的单克隆抗体KGH-R1对乳腺癌组织的特异性,为临床应用奠定基础.方法 选取56例正常乳腺组织、55例乳腺普通型导管增生、33例乳腺不典型增生、35例乳腺导管原位癌和60例浸润性乳腺癌组织,用本实验室制备的广谱抗p21ras单克隆抗体KGH-R1进行免疫组化染色,计算各样本的阳性细胞百分率和HSCORE分值,比较各组免疫反应性的差异.结果 KGH-R1单克隆抗体与71.67%(43/60)乳腺浸润癌发生免疫反应,阳性样本的平均阳性细胞率66.29%,平均HSCORE评分131.64分,KGH-R1抗体和HSCORE分值与浸润癌患者年龄、性别、淋巴结转移、ER、PR状态无关(P>0.05).与62.86%(22/35)的原位癌发生免疫反应,阳性样本的平均阳性细胞率59.18%,平均HSCORE评分86.75分.与57.58%(19/33)的不典型增生导管上皮发生免疫反应,阳性样本的平均阳性细胞率46.83%,平均HSCORE评分54.86分.与49.09%(27/55)的普通型导管增生组织发生免疫反应,阳性样本的平均阳性细胞率23.15%,平均HSCORE评分21.95分.但KGH-R1单克隆抗体仅与少数(12/56)的正常乳腺组织发生微弱免疫反应(HSCORE<30分).结论 KGH-R1单克隆抗体具有较好的乳腺癌特异性,可以进一步开发为治疗性抗体.     

抗p21Ras单克隆抗体KGH-R1在大肠良恶性病变中的免疫反应性比较

目的:评价抗p21Ras单克隆抗体(mAb)KGH-R1在大肠良恶性病变及正常黏膜组织中的免疫反应性,为该抗体的临床应用奠定基础。方法:用本实验室制备的广谱抗p21 ras mAb KGH-R1对正常大肠黏膜、大肠炎息肉、大肠低级别上皮内瘤变、大肠高级别上皮内瘤变、浸润性大肠癌及癌旁病变组织进行免疫组化染色,计算各样本的阳性细胞百分率和组织学评分(HSCORE)分值,比较各组免疫反应性的差异。结果:mAb KGH-R1与64.89%(61/94)的浸润性大肠癌发生免疫反应,阳性样本的平均阳性细胞数为97.28%,平均HSCORE评分178.98分;与60.24%(50/83)的高级别上皮内瘤变组织发生免疫反应,阳性样本的平均阳性细胞数为95.08%,平均HSCORE评分156.38分;与64.58%(31/48)的低级别上皮内瘤变组织发生免疫反应,阳性样本的平均阳性细胞数为82.52%,平均HSCORE评分103.03分;与39.97%(29/73)的炎性息肉组织发生免疫反应,阳性样本的平均阳性细胞率17.78%,平均HSCORE评分18.66分。虽然mAb KGH-R1也与46.67%(21/45)的正常大肠黏膜上皮发生免疫反应,但阳性样本的平均阳性细胞数为2.64%,平均HSCORE评分为2.64分。大肠癌与癌旁高级别上皮内瘤变组织比较,mAb KGH-R1反应阳性细胞数和HSCORE评分无统计学意义,但高于癌旁低级别上皮内瘤变组织。20/85例(23.53%)癌旁正常黏膜组织与mAb KGH-R1呈微弱阳性反应。结论:mAb KGH-R1与大肠癌具有较强的免疫反应性,有开发为治疗性抗体的可能。     

非小细胞肺癌组织中细胞凋亡与p53蛋白表达的定量研究

目的 :观察非小细胞肺癌 (NSCLC)组织中Caspases介导的细胞凋亡与p5 3蛋白表达情况 ,探讨两者之间的相关关系和意义。方法 :临床手术切除的NSCLC标本 4 4例 ,其中鳞癌 2 0例 ,腺癌 2 4例。采用Caspase特异的单克隆抗体M30Cy toDEATH免疫组化染色显示凋亡细胞 ,计算凋亡指数 (AI) ;免疫组化法检测p5 3蛋白表达 ,用显微图像分析系统测定阳性细胞百分率 (P/A)、平均光密度 (AOD)及阳性水平指数 (PLI)。结果 :NSCLC组织中M30阳性率为 2 5 0 % ,肺鳞癌组织的M 30阳性率明显高于肺腺癌组织 (P <0 0 5 )。低分化鳞癌组织的AI高于中、高分化鳞癌 (P <0 0 5 )。NSCLC组织中 p5 3蛋白阳性率为 5 2 2 %。p5 3蛋白的PLI与癌细胞AI之间无直线相关关系。 结论 :在NSCLC组织中存在Caspases介导的细胞凋亡机制 ,且与组织学类型及组织分化程度有关。p5 3蛋白表达与癌细胞凋亡无相关关系 ,提示突变型 p5 3蛋白失去了激活Cas pases而诱导细胞凋亡的作用 ,与肺癌的发生、发展、治疗及预后有关。     

肺腺癌中CD147、MMP-9和VEGF的表达及临床意义

目的观察肺腺癌中CD147、MMP-9和VEGF蛋白表达,并分析三者表达的相关性。方法采用免疫组化SP法检测90例肺腺癌、10例肺良性病变和10例相对正常肺组织中CD147、MMP-9和VEGF蛋白表达,用Imagepro Plus图像分析软件测试3种蛋白在肺腺癌中的阳性强度。结果 CD147、MMP-9和VEGF蛋白表达的阳性单位(positive unit,PU)值在肺腺癌组织中均显著高于相对正常肺组织和肺良性病变组织(P均0.001)。在90例肺腺癌中,CD147、MMP-9和VEGF的阳性率分别为88.89%(80/90)、84.44%(76/90)、80%(72/90)。CD147、MMP-9和VEGF均分别与肺腺癌组织学分化程度、淋巴结转移及TNM分期有关(P均0.001),三者与肺腺癌患者年龄、吸烟史、大体类型均无关(P均0.05)。CD147表达和MMP-9、VEGF表达均呈显著正相关(r_p=0.992,r_p=0.984,P均0.001)。结论 CD147、MMP-9和VEGF可能在肺腺癌发生、发展中起重要作用,推测CD147刺激肿瘤细胞产生MMP-9和VEGF,进而促进肺腺癌的浸润和转移,可以作为肺腺癌免疫治疗的潜在靶点。     

特异性抗体在乳腺癌和肺腺癌鉴别诊断中的应用及异常表达率的研究

目的探讨GATA3、ER、PR、TTF-1、Napsin A在乳腺癌及肺腺癌中的敏感性、特异性、异常表达率及其组合在鉴别诊断中的应用价值。方法采用免疫组化法检测GATA3、ER、PR、TTF-1、Napsin A在152例乳腺癌及116例浸润性肺腺癌中的表达,阳性细胞所占比率≥10%定义为阳性。用MedCalc软件统计各抗体及组合抗体在乳腺癌及肺腺癌鉴别诊断中的敏感性及特异性。结果 GATA3表达于87.5%的乳腺癌(管腔A/B型100%、HER-2阳性型79.2%、三阴型39.1%)及3.4%的肺腺癌(非黏液腺癌2.7%,黏液腺癌16.7%,P=0.194)。GATA3在ER阳性及阴性乳腺癌中的阳性率差异有显著性(100%vs59.6%,P0.05)。TTF-1表达于96.5%的肺腺癌(非黏液腺癌98.2%,黏液腺癌66.7%,P=0.013)及1.3%的乳腺癌(管腔A/B型0.9%、HER-2阳性型0、三阴型4.3%)。Napsin A表达于87.1%的肺腺癌(非黏液腺癌88.2%,黏液腺癌66.7%,P=0.172),乳腺癌均未见表达。肺腺癌表达ER(10.2%)、PR(1.7%)。GATA3对乳腺癌的敏感性及特异性均优于ER(87.50%vs 69.08%,96.55%vs 89.66%),而TTF-1对肺腺癌的敏感性优于Napsin A(96.55%vs 85.34%)。GATA3/TTF-1组合可鉴别87.5%的乳腺癌及93.1%的肺腺癌,而TTF-1/GATA3均阳性见于3.4%的肺腺癌及1.3%的乳腺癌。结论少数肺腺癌及乳腺癌可异常表达GATA3及TTF-1,特别是在三阴型乳腺癌及肺黏液腺癌中异常表达率偏高。在穿刺活检标本任何一种抗体都不能单独作为鉴别两者的依据。GATA3/TTF-1标记是鉴别乳腺癌及肺腺癌的首选抗体,而ER、PR及Napsin A对于鉴别诊断有较好的辅助作用。     

p16、p21在卵巢常见的上皮性肿瘤中的表达及鉴别诊断

目的探讨p16、p21在卵巢上皮性肿瘤中的表达及其鉴别诊断作用。方法采用免疫组化SP法检测314例卵巢上皮性肿瘤组织中p16、p21的表达。结果 (1)p16在卵巢浆液性腺癌、黏液性腺癌、内膜样腺癌中阳性细胞表达百分数以中位数(四分位间距)表示为80.00%(45.00%)、0.00(20.00%)、35.00%(48.75%),三者之间表达差异有统计学意义(P=0.00);p16在不同级别(良、交界性、恶性肿瘤)的卵巢浆液性肿瘤和子宫内膜样肿瘤中,随着肿瘤级别的升高,p16表达强度及阳性程度亦随之升高,差异具有统计学意义(P=0.001)。而在不同级别的卵巢黏液性肿瘤中p16的表达差异无统计学意义(P>0.05);(2)p21在不同类型卵巢上皮性肿瘤中的表达普遍较低,在卵巢浆液性腺癌、黏液性腺癌和子宫内膜样腺癌中表达阳性细胞百分数以中位数(四分位间距)表示分别为0.00(10.00%)、10.00%(15.00%)、0.00(7.50%),三者之间差异有统计学意义(P=0.000);p21在交界性肿瘤中的表达较在良性肿瘤与恶性肿瘤中的表达高,有统计学意义,卵巢恶性肿瘤与良性肿瘤之间表达差异无统计学意义。结论...     

联合检测TTF1、CK5/6、p63和napsinA在肺鳞癌和腺癌鉴别诊断中的价值

目的 探讨TTF1、CK5/6、p63和napsinA联合检测在肺鳞状细胞癌和腺癌鉴别诊断中的价值.方法 应用免疫组化EnVision法分别检测30例肺鳞状细胞癌和30例肺腺癌中TTF1、CK5/6、p63和napsinA的表达.结果 CK5/6、p63、TTF1和napsinA在肺鳞状细胞癌中的阳性率分别为96.7%、100%、20%和0,同时在肺腺癌中的阳性率分别为10%、20%、86.7%和90%.单个标志物中,CK5/6阳性和napsinA阴性对肺鳞状细胞癌与腺癌的鉴别诊断具有较高的特异性和敏感性;联合标志物检测中,p63和CK5/6共同表达阳性与TTF1和napsinA共同表达阴性在肺鳞状细胞癌的诊断中具有较高的特异性.结论 TTF1、CK5/6、p63和napsinA联合检测可鉴别肺鳞状细胞癌和腺癌,p63和CK5/6共同阳性表达与TTF1和napsinA共同阴性表达更加支持肺鳞状细胞癌,而非肺腺癌,反之亦然.     

MUC5B、Villin、P53在胆囊黏膜幽门腺化生、肠上皮化生及胆囊腺癌中的表达及意义

目的：通过观察MUC5B、Villin、P53蛋白在胆囊黏膜幽门腺化生、肠上皮化生及胆囊腺癌的表达,探讨胆囊黏膜两种化生与胆囊腺癌发生的关系。方法：收集2013年1月至2015年1月兰州市第二人民医院病理科诊断的胆囊黏膜幽门腺化生40例、肠上皮化生40例及胆囊腺癌40例,采用免疫组化方法检测MUC5B、Villin、P53在胆囊黏膜幽门腺化生、肠上皮化生及胆囊腺癌的表达。结果：MUC5B在胆囊黏膜幽门腺化生、肠上皮化生、胆囊腺癌的阳性表达率分别为95.00%(38/40)、75.00%(30/40)、27.50%(11/40);Villin在三组中的阳性表达率分别为0.00%(0/40)、87.50%(35/40)、22.50%(9/40);P53在三组中的阳性表达率分别为2.50%(1/40)、7.50%(3/40)、80.00%(32/40)。各组间比较MUC5B在幽门腺及肠上皮化生的阳性表达率明显高于胆囊腺癌,差异有统计学意义(χ2=42.754,P=0.001);Villin在肠上皮化生的阳性表达率明显高于幽门腺化生及胆囊腺癌,差异有统计学意义(χ2=71.124, P=0.001);P53在胆囊腺癌中的阳性表达率明显高于幽门腺及肠上皮化生,差异有统计学意义(χ2=71.667,P=0.001)。MUC5B、Villin在幽门腺化生、肠上皮化生及胆囊腺癌的阳性表达率递减;P53在幽门腺化生、肠上皮化生及胆囊腺癌的阳性表达率递增。结论：幽门腺及肠上皮化生可能参与了胆囊腺癌的发生。     

恶性上皮性肿瘤中CK20的表达及其临床意义

目的　研究单克隆抗体CK2 0在恶性上皮性肿瘤和卵巢转移性腺癌组织中的表达及其意义。方法　应用S P法对鼻咽非角化性癌、乳腺浸润性导管癌、肺的鳞癌和腺癌、卵巢黏液性囊腺癌、胃腺癌和结肠直肠腺癌各组总计 6 7例和 4 1例分别进行了CK2 0和CK19检测。结果　CK2 0阳性率 :肺腺癌 1/ 7(14 3% ) ,卵巢浆液性和黏液性腺癌 3/ 12 (33 3% ) ,胃腺癌 3/ 9(33 3% ) ,结肠直肠腺癌组 2 1/ 2 2 (95 5 % ) ,其他癌组织均呈阴性。结肠直肠腺癌组组与其他各组间比较差异有显著性 (P <0 0 1)。CK19在上述 4 1例癌组织中均呈强阳性表达。结论　CK2 0表达对鉴别结肠腺癌和直肠腺癌与肺腺癌和乳腺浸润性导管癌具有高度特异性和较高的敏感性 ;CK2 0高表达对鉴别卵巢原发性腺癌与卵巢的结肠腺癌或直肠腺癌转移具有一定的意义     

D2-40在肺非典型腺瘤样增生及腺癌中的辅助诊断价值

目的 探讨D2-40在肺的前驱腺体病变[包括非典型腺瘤样增生(atypical adenomatous hyperplasia, AAH)和原位腺癌(adenocarcinoma in situ, AIS)]、微浸润性腺癌(minimally invasive adenocarcinoma, MIA)及浸润性腺癌中的辅助诊断价值。方法 收集AAH、AIS、MIA及浸润性腺癌组织各20例,每例肿瘤均有癌旁正常肺组织作为对照,采用免疫组化EnVision两步法检测D2-40表达,并复习相关文献。结果 D2-40在所有正常肺组织肺泡上皮中均100%(80/80)阳性,且呈胞质连续性强着色;在20例AAH、AIS和MIA中的阳性率分别为85%(17/20)、50%(10/20)和45%(9/20),其着色强度呈微弱～中等,染色模式呈现不连续或不等间隔,其中MIA中浸润性成分阳性率为30%(6/20),贴壁成分阳性率为45%(9/20);20例浸润性腺癌中仅1例微弱着色,阳性率为5%(1/20)。统计分析显示,AAH、AIS、MIA中D2-40表达分别与浸润性腺癌和正常肺组织相比,差异均有统计学...     

Expression of Lewis X-related antigens in adenocarcinomas of lung

Fifty primary lung adenocarcinomas were examined immunohistochemically using monoclonal antibodies to determine changes in the expression of N-acetyl-lactosamine (blood group type-2 chain), Le^x, Le^Y and sialyl Le^x-i. These antigens were expressed in 60%, 70%, 90% and 94% of carcinomas, respectively; in 8%, 12%, 56% and 86% of normal broncho-bronchiolar epithelium; and in 32%, 0%, 100% and 0% of normal alveolar epithelium. The greater the complexity of the antigenic structure, the greater the incidence of positive staining in the adenocarcinomas. Although the more complex antigens such as sialyl Le^x-i and Le^Y have also been demonstrated in the sera of lung cancer patients, they were not always cancer-selective in our immunohistochemical study. In contrast, the less complex antigens such as N-acetyl-lactosamine (type-2 chain) and Le^x seem to be cancer-selective, as they showed low positivity in normal lung tissue.     

Neurotrophins and neurotrophin receptors in human lung cancer.

The expression of neurotrophins (NTs) and related high- and low-affinity receptors was studied in surgical samples of histologically diagnosed human tumors of the lower respiratory tract. The experiment was conducted with 30 non-small cell lung cancer specimens and in eight small cell lung cancer specimens by Western blot analysis and immunohistochemistry to assess expression and distribution of NT and NT receptor proteins in tissues examined. Immunoblots of homogenates from human tumors displayed binding of anti-nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and NT-3 antibodies as well as of anti-tyrosine-specific protein kinase (Trk) A, TrkB, and TrkC receptor antibodies, with similar migration characteristics than those displayed by human beta-NGF and proteins from rat brain. A specific immunoreactivity for NTs and NT receptors was demonstrated in vessel walls, stromal fibroblasts, immune cells, and sometimes within neoplastic cell bodies. Approximately 33% of bronchioloalveolar carcinomas exhibited a strong membrane NGF and TrkA immunoreactivity, whereas 46% adenocarcinomas expressed an intense TrkA immunoreactivity but a weak immunostaining for NGF within tumor cells. Moreover, squamous cell carcinomas developed an intense TrkA immunoreactivity only within stroma surrounding neoplastic cells. A faint BDNF and TrkB immunoreactivity was documented in adenocarcinomas, squamous cell carcinomas, and small cell lung cancers. NT-3 and its corresponding TrkC receptor were found in a small number of squamous cell carcinomas within large-size tumor cells. No expression of low-affinity p75 receptor protein was found in tumor cells. The detection of NTs and NT receptor proteins in tumors of the lower respiratory tract suggests that NTs may be involved in controlling growth and differentiation of human lung cancer and/or influencing tumor behavior.     

OV-TL 12/30 (keratin 7 antibody) is a marker of glandular differentiation in lung cancer

The immunoreactivity of OV-TL 12/30, a monoclonal antibody to keratin 7 was investigated on paraffin-embedded human lung cancer tissues of 61 patients. A modified AEC-immunoperoxidase method with pepsin pre-digestion was used. In normal lung tissue keratin 7 was found in bronchial and bronchiolar epithelium, pneumocytes and compound glands. Squamous metaplasia of the bronchial tree was negative. All 24 squamous cell carcinomas were negative irrespective of grade of differentiation. All differentiation grades of 20 adenocarcinomas including bronchioalveolar carcinomas were positive. Since six large cell anaplastic carcinomas did not react with keratin 7 antibody these tumours are considered to be of squamous cell rather than adenocarcinomatous origin. Small cell anaplastic carcinomas were negative in 10 of 11 cases. Our study demonstrates that this keratin 7 antibody is useful in differentiating between squamous cell carcinoma and adenocarcinoma of the lung and it may be particularly useful in making the correct diagnosis in small lung biopsy specimens.     

Preservation of cluster 1 small cell lung cancer antigen in zinc-formalin fixative and its application to immunohistological diagnosis

Monoclonal antibodies reactive with cluster 1 small cell lung cancer antigen have been shown to be useful for the distinction of small cell from non-small cell tumours. In previous studies the antibodies have been applied to frozen sections and cold acetone-fixed tissues. However, one of three monoclonal antibodies that we produced, NCC-LU-243, reacted with some small cell lung carcinomas fixed in formalin solution and embedded in paraffin. The addition of zinc sulphate to the formalin solution at a concentration of 2% (v/w) greatly improved antigen immunoreactivity, and reactivity was retained even after prolonged fixation. Occasionally, immunoreactivity was present in a poorly differentiated adenocarcinoma with rosette-like structures. The monoclonal antibody NCC-LU-243 is thus of considerable potential value in the immunohistochemical diagnosis of small cell lung cancers.     

Monoclonal anti-CEA antibodies in the discrimination between primary pulmonary adenocarcinoma and colon carcinoma metastatic to the lung

Lung metastases from colon adenocarcinoma are often difficult to differentiate from primary lung adenocarcinoma. We studied the diagnostic value of a polyclonal anti-CEA antiserum and two monoclonal anti-CEA antibodies (B18, D14) which define antigens overexpressed in colon carcinoma. Autopsy material from 20 patients with colon carcinoma and lung metastases and 20 specimens from patients with primary lung adenocarcinoma were retrieved, stained, and interpreted without knowledge of the origin of the lung tumor. Colon carcinomas, lung metastases and lung primaries stained positively with polyclonal anti-CEA in 90-100% of cases. D14 stained 75% of colonic metastases and 70% of primary lung adenocarcinomas, whereas 95% of colon primaries were positive. Sixty-five percent of colon primaries and 50% of their metastases were positive with B18, whereas 45% of lung primaries were positive. The frequency of B18 positivity was significantly greater in those colon primaries that were surgically derived (7/9, 78%) compared with their autopsy-derived lung metastases (2/9, 22%) (P less than 0.05). Similarly, D14 staining in surgically derived colon primaries (9/9, 100%) was significantly greater than their autopsy-derived lung metastases (5/9, 56%) (P less than 0.05). In surgical/biopsy-derived tissues 9/9 colonic primaries were D14-positive, whereas only 1 of 6 lung primaries was positive (P = 0.002). We conclude that D14 and polyclonal anti-CEA both stain the majority of colon adenocarcinomas and that changes associated with prolonged fixation may reduce the positivity rate with both B18 and D14 monoclonal antibodies. All three antibodies stain autopsy-derived tissue from primary lung cancer to a significant degree.(ABSTRACT TRUNCATED AT 250 WORDS)     

Immunohistochemical and immunoblot analyses of ras-p21 expression in lung carcinomas

Immunohistochemical and immunoblot analyses were performed on 49 cases of surgically resected primary lung carcinoma to examine the expression of ras oncogene product using monoclonal antibodies to ras-p21. Two different monoclonal antibodies, NCC-RAS-001 and RAP-5 were used for immunohistochemical study. Cancer cells of 16 cases (33%) and 15 cases (31%) were strongly positive for NCC-RAS-001 and RAP-5, respectively. The staining pattern of antibodies was heterogenous among cancer cells, even in the same case. Among various histologic type of lung cancers, squamous cell carcinomas and well-differentiated adenocarcinomas had a tendency to react more intensively than other histologic types of carcinoma. Immunoblot analysis using monoclonal antibody NCC-RAS-004 revealed the presence of ras-p21 not only in cancer tissues but also in non-cancerous tissues in all cases analysed. In 13 cases (27%), cancer tissue expressed more than twice as much ras-p21 as non-cancerous tissues. Our study showed that the over-expression of ras-p21 in lung carcinomas compared with non-cancerous tissues was a relatively common phenomenon, especially in squamous cell carcinomas and adenocarcinomas.