Immunohistochemical localization of pulmonary surfactant apoproteins in various lung tumors. Special reference to nonmucus producing lung adenocarcinomas

Eighty-nine primary lung carcinomas and 23 metastatic lung tumors were immunohistochemically studied for the expression of pulmonary surfactant apoproteins, by using monoclonal (PE-10) and polyclonal antibodies. Surfactant apoprotein was demonstrated in the cytoplasm and/or nuclear inclusion bodies of only primary lung adenocarcinomas (36 of 75 cases), not in any other histologic type of primary lung carcinoma or in metastatic lung tumors. In primary lung adenocarcinoma, although typical type II pneumocyte type adenocarcinoma was not included in the current series, the majority of surfactant apoprotein-positive single cell type tumors were of the Clara cell type, with a single bronchial surface epithelial cell type, according to the light microscopic subclassification of adenocarcinoma cells. The Clara cell type adenocarcinomas could at times be distinguished only with difficulty from adenocarcinoma of type II pneumocyte type. Normal and hyperplastic type II pneumocytes were of course positive for surfactant apoprotein in the cytoplasm. However, none of the positive cells could definitely be identified as Clara cells in non-neoplastic lungs. The findings obtained in this study indicate that surfactant apoprotein is a good marker to distinguish adenocarcinoma of the lung from other histologic types of lung cancer and from neoplasms metastatic to the lung, and that type II pneumocytes and Clara cells, non-neoplastic and neoplastic, are morphologically and functionally closely related and might belong to the same cell lineage.     

Nm23 GENE EXPRESSION AND ITS CORRELATION WITH LYMPH NODE METASTASIS IN HUMAN LUNG CANCER

雷文东，张汝刚，阎水忠，王秀琴，牟巨伟，张大为，吴Ｎｍ２３ＧＥＮＥＥＸＰＲＥＳＳＩＯＮＡＮＤＩＴＳＣＯＲＲＥＬＡＴＩＯＮＷＩＴＨＬＹＭＰＨＮＯＤＥＭＥＴＡＳＴＡＳＩＳＩＮＨＵＭＡＮＬＵＮＧＣＡＮＣＥＲ￥ＬｅｉＷｅｎｄｏｎｇ；ＺｈａｎｇＲｏｕｇｉｎｇ；...     

Immunohistochemical Analysis of Cathepsin B Expression in Human Lung Adenocarcinoma: The Role in Cancer Progression

Production of cathepsin B by tumor cells has been linked to metastatic potential in several experimental models. Sections of 95 primary lung adcnocarcinomas were examined for expression of cathepsin B using a standard avidin-biotin immunohistochemical technique. Staining for cathepsin B was observed in 22.1% of all cases and 28.0% of those of the Clara cell type. In Clara cell adenocarcinomas, cathepsin B expression correlated with positive lymph node status, presence of distant metastases, and poor prognosis ( P <0.05). However, no correlation with clinical outcome was observed in other cell types. Our data suggest that cathepsin B may be involved in invasion and metastasis in Clara cell lung adenocarcinoma.     

Immunohistochemical analysis of nm23 gene product/NDP kinase expression in pulmonary adenocarcinoma: lack of prognostic value.

Levels of nm23 gene product/nucleoside diphosphate kinase (NDP kinase) expression have been demonstrated to correlate inversely with metastatic potential in several tumours, indicating that this could be a useful tool as a prognostic indicator. Using an antibody to NDP kinase, levels of nm23 gene product/NDP kinase expression in pulmonary adenocarcinoma were examined immunohistochemically. Of 88 patients tested, 39 (44%; Group B) showed strong immunoreactivity for NDP kinase in most of cancer cells within the tumour tissues, while 49 (56%; Group A) contained few or no NDP kinase-positive cancer cells. Nm23 gene product/NDP kinase was expressed independently of clinicopathological factors, and unexpectedly, no correlation of survival rates between both Groups could be demonstrated. Thus, in pulmonary adenocarcinoma, levels of nm23 gene product/NDP kinase expression may lack prognostic value.     

nm23和p53基因与肺腺癌生物学特性的关系

目的：检测肺腺癌组织中nm23表达和p53基因外显子突变情况,寻找有助于准确预测局部肿瘤进展、分型和预后情况的临床参照指标。方法：应用PCR技术检测31例肺腺癌术后存档蜡块组织中nm23表达及p53基因外显子突变情况,并与肺腺癌的病理及临床指标进行相关性分析。结果：nm23表达及p53基因突变率分别为38．7％（12／31）及48．4％（15／31）。nm23表达和p53基因突变与临床分期相关,P值分别为0．005、0．037。结论：nm23表达和p53基因突变与肺癌临床分期和生存相关,在判断肺腺癌细胞增殖、分化程度、恶性程度及预后转归方面有一定价值,可作为判断肺腺癌预后的参考指标,指导高危患者的进一步治疗。     

nm23基因在人胰腺癌中的表达及预后的关系

目的探讨nm23/NDPK癌基因在胰腺癌组织中的表达及其临床意义.方法采用免疫组化SP法,对40例胰腺癌组织及10例正常胰腺组织进行nm23/NDPK癌基因检测.结果40例胰腺癌组织中有26例nm23/NDPK表达阳性,占65%;10例正常胰腺组织中仅3例nm23/NDPK表达阳性,占30%,两者有显著差异(P＜0.05).低分化腺癌nm23/NDPK阳性表达率明显高于高分化胰腺癌(10/11,90.9%;2/8,25%;P＜0.05).其nm23/NDPK表达阳性与淋巴结转移密切相关(10/14,71.4%;6/19 31.5%;P＜0.05).提示胰腺癌nm23/NDPK表达与淋巴结转移及肿瘤侵袭性呈正相关,与癌组织的分化程度呈负相关.结论nm23/NDPK表达可作为胰腺癌恶性程度及预后不良的生物学指标之一.     

nm23-H1基因对人肺癌细胞中细胞外信号调节激酶活性的影响

目的探讨肿瘤转移抑制基因nm23-H1对人高转移大细胞肺癌细胞株L9981中细胞外信号调节激酶ERK1/2活性的影响.方法应用特异性识别总ERK1/2(p44/42 MAP kinase)和双磷酸化ERK1/2(phospho-p44/42 MAP kinase)的抗体及蛋白印迹法(Western blot),检测L9981(缺失nm23-H1基因的原代肺癌细胞株)、L9981-nm23-H1(转染了nm23-H1基因的L9981细胞株)、L9981-PLXSN(转染了空载体的L9981细胞株)中总ERK1/2和磷酸化ERK1/2的水平.磷酸化ERK1/2活性应用非放射性免疫沉淀和Western blot法以及p44/42 MAP kinase分析试剂盒予以检测.结果 L9981-nm23-H1细胞株中磷酸化ERK1/2的水平,以及ERK1/2活性均显著低于L9981细胞株和L9981-PLXSN细胞株(P<0.01),L9981和L9981-PLXSN细胞株间磷酸化ERK1/2水平和ERK1/2活性比较均无显著性差异(P＞0.05).三个细胞株间总ERK1/2水平比较均无显著性差异(P>0.05).结论 nm23-H1基因可明显靶向地抑制人高转移肺癌细胞株L9981中ERK1/2的转录表达和ERK1/2的活性.推测nm23-H1基因的作用机制可能与其抑制了MAPK/ERK信号传导通路有关.     

慢病毒介导的稳定沉默nm23-H1基因的肺癌细胞株的建立及生物学行为改变

背景与目的 nm23-H1基因是重要的肿瘤转移抑制基因。前期研究发现利用化学合成的小干扰RNA(small interfering RNA,siRNA)抑制nm23-H1基因的表达可明显增强肺癌细胞的侵袭力。为了进一步研究nm23-H1基因沉默后的分子生物学机制,本研究利用慢病毒介导的短发夹RNA(short hairpin RNA,shRNA)建立nm23-H1基因稳定沉默的肺癌细胞株。方法将表达特异性抑制nm23-H1基因shRNA的慢病毒转染人大细胞肺癌细胞株NL9980和肺腺癌细胞株A549,通过嘌呤霉素筛选出稳定转染细胞株。逆转录PCR、定量PCR及Western blot法检测nm23-H1基因表达,并通过shRNA抵抗的nm23-H1基因重组质粒转染拯救实验验证,侵袭小室实验检测侵袭力改变。结果逆转录PCR、定量PCR和Western blot法检测稳定转染细胞株NL9980-99和A549-99中nm23-H1基因在mRNA和蛋白水平表达均明显降低;shRNA抵抗的nm23-H1基因重组质粒转染拯救实验重现nm23-H1的正常表达;侵袭小室实验显示NL9980-99和A549-99细胞侵袭力明显增强。结论成功建立nm23-H1基因稳定沉默的人大细胞肺癌细胞株NL9980-99和人肺腺癌细胞株A549-99,nm23-H1基因沉默后使NL9980-99和A549-99细胞的侵袭力明显增强。     

nm23蛋白在非小细胞肺癌患者肺癌组织和淋巴结表达的临床意义

目的探讨ｎｍ２３基因与肺癌转移的关系。方法采用链霉抗生物素蛋白－过氧化物酶（Ｓ－Ｐ）快速免疫组织化学法检测４９例非小细胞肺癌（ＮＳＣＬＣ）患者肺癌组织和４９份配对淋巴结标本ｎｍ２３基因产物－核苷二磷酸激酶（ＮＤＰＫ／ｎｍ２３）。结果ＮＤＰＫ／ｎｍ２３在ＮＳＣＬＣ的肺癌组织表达为５７．１４％（２８／４９），其中鳞癌为６０．８７％（１４／２３），腺癌为５３．８５％（１４／２６）。２６例伴淋巴结转移的肺癌标本中阳性１６例（６１．５４％）。ＮＤＰＫ／ｎｍ２３表达与淋巴结转移没有负相关，在淋巴结转移灶有较高表达（５３．８５％，１４／２６）。结论ｎｍ２３基因在肺癌的发生、转移中可能起着不同的作用。     

肺腺癌及鳞癌MDR1-mRNA和MRP-mRNA表达的研究

观察３０例肺癌组织及癌肺组织ＭＤＲ１－ｍＲＮＡ及ＭＲＰ－ｍＲＮＡ的表达。方法：ＲＴ－ＰＣＲ方法。结果：ＭＤＲ１－ｍＲＮＡ在肺癌组织及癌旁肺组织的表达阳性率分别为４０．０％及１６．６７％（Ｐ＝０．０４５）,ＭＤＲ１－ｍＲＮＡ的表达与细胞分化程度,临床分期及病理类型无关;ＭＲＰ－ｍＲＮＡ在肺癌主癌旁肺组织的表达分别为４３．３３％及２６．６７％,ＭＲＰ＝ｍＲＮＡ的表达与细胞分化程度有关,低分者ＭＲＰ的表     

非小细胞肺癌中乳酸脱氢酶A/B的表达特点及其与肿瘤临床病理参数的关系

目的:探讨非小细胞肺癌中乳酸脱氢酶A/B（lactate dehydrogenase A/B,LDHA/LDHB）表达特征及其与肿瘤临床病理参数的关系。方法:从GEO、TCGA数据库获取非小细胞肺癌、肺腺癌、肺鳞癌基因表达数据及临床信息,通过非参数检验分析LDHA/LDHB在癌组织和正常肺组织的表达差异,通过Kaplan-Meier法和Log-rank test评估癌患者生存状况,采用chi-square test和Spearman's test分析LDHA/LDHB表达与临床特征及肿瘤恶性生物学标志物的关系。结果:LDHA/LDHB在非小细胞肺癌、肺腺癌和肺鳞癌中均高表达。在肺腺癌中,高表达LDHA的病人具有更短的总生存期和无进展生存期,高表达LDHB的病人总生存期更短,LDHA与LDHB均高表达的患者总生存期和无进展生存期皆最短。在肺鳞癌中,LDHA/LDHB表达与生存期无关。在肺腺癌中,LDHA表达与淋巴结转移、肿瘤分期相关,LDHB表达与肿瘤分期相关。在肺鳞癌中,LDHA表达与临床特征关联均不明显,LDHB表达与年龄、淋巴结转移相关。在肺腺癌中,LDHA表达水平与PCNA、Ki67、CDH2正相关,与CDH1无关,而LDHB表达水平与PCNA、Ki67正相关,与CDH2负相关,与CDH1无关。结论:LDHA与LDHB表达和肺腺癌恶性程度相关,高表达LDHA与LDHB可能预示着较差的预后及临床特征。     

Northern印迹杂交分析nm23基因在人肺癌中的表达研究

目的探讨ｎｍ２３基因表达在人肺癌中的作用。方法通过Ｎｏｒｔｈｅｒｎ印迹杂交,检测４０例人肺癌组织和１９例非癌肺组织的ｎｍ２３Ｈ１和ｎｍ２３Ｈ２ｍＲＮＡ表达,采用地高辛标记和检测系统显示杂交信号,并分析ｍＲＮＡ表达与肺癌临床特征的关系。结果低分化鳞癌的ｎｍ２３Ｈ２ｍＲＮＡ表达较中高分化鳞癌显著降低（Ｐ＜０．０１）,小细胞肺癌的ｎｍ２３Ｈ１和ｎｍ２３Ｈ２ｍＲＮＡ表达较肺鳞癌明显降低。但在有或无淋巴结转移的原发癌灶组织间,以及肺癌的临床分期间,ｎｍ２３基因ｍＲＮＡ表达差异无显著性（Ｐ＞０．０５）。结论ｎｍ２３基因ｍＲＮＡ表达与肺癌的组织分化有关,未发现其在肺癌中的癌转移抑制作用。     

Overexpression of nm23-H2/NDP kinase B in a human oral squamous cell carcinoma cell line results in reduced metastasis, differentiated phenotype in the metastatic site, and growth factor-independent proliferative activity in culture.

The metastasis suppressor activity of nm23/nucleoside diphosphate (NDP) kinase was assessed using human oral squamous cell carcinoma (SCC) cell lines. When the expression of nm23/NDP kinase was compared among several SCC cell lines, nm23-H2/NDP kinase B gene product, but not nm23-HI/NDP kinase A gene product, was reduced in the metastatic cells. Transfection of nm23-H2 into the metastatic SCC cell line LMF4 caused reduction in the lung metastasis in an experimental metastasis assay. A histological analysis of the pulmonary metastatic foci revealed that although foci of the control clones were composed of anaplastic squamous cells, those of the nm23-H2-transfected clones consisted of mostly well-differentiated cells mimicking normal stratified epithelial constitution. The transfected cells were morphologically indistinguishable from the control ones in culture, but they differed from each other in that the former cells proliferated faster than the latter, became less serum dependent, and lost responsiveness to growth factors such as platelet-derived growth factor, insulin-like growth factor I, and insulin, although both clones retained sensitivity to transferrin. These results demonstrate that nm23-H2 protein does have metastasis suppressor activity for human SCC cells and suggest that this activity may be elicited by modulating growth and/or differentiation potential in response to environmental factors.     

nm23基因在结肠癌中表达及其临床意义

目的:阐明nm23基因产物在结肠癌中表达意义及其与淋巴结转移、预后的关系。方法:用免疫组化方法研究53例结肠癌中nm23基因产物表达。结果:53例结肠癌病例中nm23基因表达阴性29例占54.7%,阳性24例占36.3%。29例阴性表达病例中,发生淋巴结转移23例占79.3%,5年生存率为31.0%(9/29)。低表达的37例均属于C2、D期。低分化腺癌和黏液腺癌占36例。提示nm23基因的低表达与结肠癌的组织学分型、局部淋巴结转移、临床分期及预后有关系。而与肿瘤的大小、部位无关。结论:检测结肠癌组织中nm23基因产物表达可预测肿瘤的转移、复发及预后。     

P2RY6在非小细胞肺癌中的表达和临床意义分析

  目的  探讨P2RY6在非小细胞肺癌（non-small cell lung cancer,NSCLC）患者中的表达及临床意义。  方法  在基因表达集（Gene Expression Omnibus,GEO）和癌症基因组图谱计划（The Cancer Genome Atlas Program,TCGA）数据库中获取多个NSCLC、肺腺癌和肺鳞癌的基因表达以及临床信息数据集。使用非参数检验分析癌组织与邻近正常组织的P2RY6表达水平差异,并且通过免疫组织化学研究肺鳞癌及肺腺癌组织和正常组织的P2RY6蛋白表达情况。使用χ2检验分析P2RY6表达与肺腺癌、肺鳞癌患者临床特征之间的相关性。使用Kaplan-Meier方法和Log-rank检验评估肺腺癌和肺鳞癌P2RY6表达水平与总生存期和无进展生存期之间的关系。使用Cox比例风险回归模型进一步评估P2RY6表达量对肺鳞癌患者总生存期和无进展生存期的预测效能。  结果  P2RY6在NSCLC、肺腺癌和肺鳞癌中均高表达。在肺腺癌患者中,P2RY6表达与生存无关,而与性别有关。在P2RY6高表达的肺鳞癌患者中,总生存期和无进展生存期较短,P2RY6高表达是独立危险因素。  结论  P2RY6与肺鳞癌患者的生存相关,P2RY6高表达预示着肺鳞癌患者总生存期与无进展生存期较短,是预后不良的独立危险因素。      

Podocalyxin influences malignant potential by controlling epithelial–mesenchymal transition in lung adenocarcinoma

Epithelial–mesenchymal transition (EMT) plays an important role in the progression of lung carcinoma. Podocalyxin (PODXL), which belongs to the CD34 family and regulates cell morphology, has been linked to EMT in lung cancer, and PODXL overexpression is associated with poor prognosis in several different classes of cancers. The aim of this study was to clarify the role of PODXL overexpression in EMT in lung cancer, and to determine the prognostic value of PODXL overexpression in tumors from lung cancer patients. The morphology, EMT marker expression, and migration and invasion abilities of engineered A549 PODXL‐knockdown (KD) or PODXL‐overexpression (OE) lung adenocarcinoma cells were examined. PODXL expression levels were assessed by immunohistochemistry in 114 human clinical lung adenocarcinoma specimens and correlated with clinical outcomes. PODXL‐KD cells were epithelial in shape, whereas PODXL‐OE cells displayed mesenchymal morphology. Epithelial markers were upregulated in PODXL‐KD cells and downregulated in PODXL‐OE cells, whereas mesenchymal markers were downregulated in the former and upregulated in the latter. A highly selective inhibitor of phosphatidylinositol 3‐kinase‐Akt signaling attenuated EMT of PODXL‐OE cells, while a transforming growth factor inhibitor did not, suggesting that PODXL induces EMT of lung adenocarcinoma cells via the phosphatidylinositol 3‐kinase pathway. In lung adenocarcinoma clinical specimens, PODXL expression was detected in minimally invasive and invasive adenocarcinoma, but not in non‐invasive adenocarcinoma. Disease free survival and cancer‐specific survival were significantly worse for patients whose tumors overexpressed PODXL. PODXL overexpression induces EMT in lung adenocarcinoma and contributes to tumor progression.     

肺癌的淋巴结转移与nm23—H1,PCNA表达的关系

目的研究ｎｍ２３－Ｈ１,ＰＣＮＡ在肺癌中的表达与肺癌转移的关系。方法应用免疫组化ＳＰ法检测４７例肺癌中ｎｍ２３－Ｈ１,ＰＣＮＡ的表达。结果原发性肺癌中ｎｍ２３－Ｈ１,ＰＣＮＡ的阳性率分别为５１．０６％和６５．９６％。其中ｎｍ２３－Ｈ１在鳞癌中的阳性率４３．３３％（１３／３０）,腺癌６４．７０％（１１／１７）。在鳞癌中无肺门及纵膈淋巴结转移阳性率６２．５％（１０／１６）,有转移组阳性率２１．４２％（３／１４）,差异有显著性（Ｐ＜０．００１）,ｎｍ２３－Ｈ１表达与鳞癌的淋巴结转移呈负相关。在ＰＣＮＡ表达中有淋巴结转移阳性率８２．６％（１９／２３）,无转移组５４．１６％（１３／２４）（Ｐ＜０．００１）,同时低分化癌的阳性率８８．８８％（８／９）明显高于高分化癌４３．７５％（７／１６）,（Ｐ＜０．００１）。结论ｎｍ２３－Ｈ１,ＰＣＮＡ的表达对肺癌予后评估有一定意义。     

p53、C-erbB-2和nm23基因表达在非小细胞肺癌中的临床意义

目的：探讨p53．C-erbB-2和nm23基因在非小细胞肺癌（NSCLC）组织中表达的临床意义。方法：组织学或细胞学证实的NSCLC89例,采用免疫组化法检测p53、C-erbB-2和nm23蛋白表达,并取癌旁正常肺组织87例作为对照。结果：肺癌组织p53和C-erbB-2蛋白表达率分别为60％和67％,均高于正常肺组织5％和3％（P〈0．05）;nm23蛋白表达为67％,则显著低于正常肺组织93％（P〈0．05）：p53蛋白表达与肺癌组织的病理类型、原发肿瘤范围（T）和肿瘤分化程度有显著的相关性（P〈0．05）,与患者的性别、年龄、疾病分期（TNM）、淋巴结受侵范围（N）和远处转移状况（M）没有明显的相关性。C-erbB-2蛋白表达与患者的性别、年龄、原发肿瘤大小（T）、淋巴结受侵范围（N）和远处转移状况（M）、疾病分期（TNM）及肿瘤分化程度均没有明显的相关性。nm23蛋白表达与肿瘤的淋巴结受侵范围（N）、远处转移状况（M）和疾病分期（TNM）存在显著相关性（P〈0．05）,与患者的性别、年龄、原发肿瘤大小（T）及分化程度没有显著相关性。结论：p53基因编码的蛋白在NSCLC表达增高及nm23基冈编码的蛋白表达降低,提示p53和nm23基因在NSCLC的发生发展、病理特征和转移中可能起重要作用。     

山东地区非小细胞肺癌分子靶向治疗驱动基因表达情况及临床特征分析

背景与目的分子生物学靶向治疗已逐渐成为非小细胞肺癌（non-small cell lung cancer, NSCLC）的一个重要治疗手段,本研究通过分析山东地区NSCLC多种驱动基因表达情况及临床病理特征,为筛选分子靶向治疗目标人群提供理论依据。方法采用荧光探针PCR法检测表皮生长因子受体（epidermal growth factor receptor, EGFR）、棘皮动物微管相关蛋白4-间变性淋巴瘤激酶（echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase, EML4-ALK）、肉瘤致癌因子-受体酪氨酸激酶（ROS proto-oncogene 1, receptor tyrosine kinase, ROS1）、鼠类肉瘤病毒癌基因（Kirsten rat sarcoma viral oncgene, KARS）基因表达情况,回顾性分析阳性病例的临床病理特征。结果 EGFR基因突变阳性率为36.70%,主要为19、21外显子突变,突变人群主要为女性、腺癌、不吸烟患者,组间差异有统计学意义。EML4-ALK融合基因重排阳性率为9.37%。人群特征主要为60岁以下不吸烟人群,组间差异有统计学意义,基因突变与病理类型和性别间无明显差异。ROS1融合基因重排阳性率为3.67%,均为60岁以下患者,组间差异有统计学意义。23份病例标本开展KRAS基因检测,阳性标本数2例,阳性率为8.70%。2份阳性标本均为60岁以上病例,男女各占1例,病理类型均为腺癌,均无吸烟史。此外,未发现有两种基因同时突变的病例。结论 EGFR、EML4-ALK、ROS1、KARS基因在NSCLC患者中存在较高的突变率,且具有不同的人群特征,在选择靶向治疗人群中具有重要意义。