Adrenomedullin expression does not correlate with survival in lung cancer

It is suggested that adrenomedullin (AM) plays a role in lung carcinogenesis although, to confirm this suggestion, further clinical studies are needed to determine its relationship with prognosis in lung cancer. Archived 50 paraffin-embedded tumor samples of the lung were retrospectively evaluated for AM expression by immunohistochemistry and analyzed for a possible correlation with patient characteristics and survival. Quantitation of immunoreactivity was accomplished using an immunohistochemical scoring system. The pulmonary resection specimens contained 22 squamous cell carcinomas, 15 adenocarcinomas, and 13 small cell carcinomas. Non-small cell carcinomas of the lung were more likely to express AM than small cell carcinomas of the lung. Ninety-one percent of squamous cell carcinomas and 87% of adenocarcinomas expressed AM at a moderate to strong level and grade2–4 (30-100%), which were significantly higher from the non-neo-plastic lung tissue. Twenty-three percent of small cell carcinomas of lung expressed AM. Interestingly, AM immunoreactivity was essentially weak and grade 1 (<%30) in this group. AM expression is upregulated in non-small cell carcinomas of the lung, whereas it is downregulated in small cell carcinomas and non-neo-plastic lung tissues. AM expression did not show any correlation with the differentiation of the tumor, the stage of cancer, and the overall survival of patients. These results did not support the role of adrenomedullin as an independent survival factor for lung cancer. However, AM inhibition in conjunction with other anti-angiogenic agents may be useful in the prevention and treatment of malignancies.     

Integrin (alpha 6/beta 4) expression in human lung cancer as monitored by specific monoclonal antibodies

In this study, the expression of the alpha 6/beta 4 integrin complex was analyzed in human lung carcinomas both in vitro and in vivo, using two monoclonal antibodies which recognize the integrin subunits alpha 6 (Mab 135-13C) and beta 4 (Mab 439-9B). Immunoprecipitation patterns obtained from established human lung carcinoma cell lines demonstrated that the alpha 6 and the beta 4 subunits were differentially expressed in carcinomas of different types. The alpha 6 subunit was expressed in all the cell lines tested (squamous cell carcinoma A431, adenocarcinoma A549, large cell carcinoma DG3, and small cell carcinoma AE2). The beta 4 subunit was expressed in non-small cell cancer lines but was not detectable in the small cell cancer line tested. Using a quantitative two-site assay, we measured the concentration of the alpha 6/beta 4 integrin in matched biopsies from primary lung tumors and from normal lung. These studies confirmed that the complex was differentially expressed in non-small versus small cell lung cancers and that it was also detectable in lysates from normal lung at low levels. The highest levels of alpha 6/beta 4 were found in moderately differentiated squamous cell carcinomas. By immunohistochemistry, the beta 4 subunit was detectable in all the squamous cell carcinoma and adenocarcinomas tested (a total of 59), but not in 10 small cell cancers. The patterns of immunoreactivity were consistent with the expected distribution of membrane glycoproteins and, in some squamous cell carcinomas, were suggestive of the localization displayed by molecules involved in carcinoma-stroma interaction. Immunohistochemical staining indicated that beta 4 was also expressed in specific types of nonrespiratory pulmonary epithelial cells.     

High frequency of epidermal growth factor receptor mutations with complex patterns in non-small cell lung cancers related to gefitinib responsiveness in Taiwan.

PURPOSE: Epidermal growth factor receptor (EGFR) mutations related to gefitinib responsiveness in non-small cell lung cancer have been found recently. Detection of EGFR mutations has become an important issue for therapeutic decision-making in non-small cell lung cancer. EXPERIMENTAL DESIGN: Mutational analysis of the kinase domain of EGFR coding sequence was done on 101 fresh frozen tumor tissues from patients without prior gefitinib treatment and 16 paraffin-embedded tumor tissues from patients treated with gefitinib. Detection of phosphorylated EGFR by immunoblot was also done on frozen tumor tissues. RESULTS: The 101 non-small cell lung cancer tumor specimens include 69 adenocarcinomas, 24 squamous cell carcinomas, and 8 other types of non-small cell lung cancers. Mutation(s) in the kinase domain (exon 18 to exon 21) of the EGFR gene were identified in 39 patients. All of the mutations occurred in adenocarcinoma, except one that was in an adenosquamous carcinoma. The mutation rate in adenocarcinoma was 55% (38 of 69). For the 16 patients treated with gefitinib, 7 of the 9 responders had EGFR mutations, and only 1 of the 7 nonresponders had mutations, which included a nonsense mutation. The mutations seem to be complex in that altogether 23 different mutations were observed, and 9 tumors carried 2 mutations. CONCLUSIONS: Data from our study would predict a higher gefitinib response rate in lung adenocarcinoma patients in Chinese and, possibly, other East Asian populations. The tight association with adenocarcinoma and the high frequency of mutations raise the possibility that EGFR mutations play an important role in the tumorigenesis of adenocarcinoma of lung, especially in East Asians.     

非小细胞肺癌中WWOX蛋白及ErbB2蛋白的联合检测

 目的 探讨WWOX(WW domain-containing oxidoreductase)基因在非小细胞肺癌组织中的表达及与ErbB2蛋白之间的关系。 方法 用免疫组化方法检测50例肺鳞癌,31例肺腺癌及20例癌旁正常组织中WWOX基因蛋白的表达,并分析其与各临床病理指标及ErbB2蛋白之间的相关性。 结果 WWOX基因在72.8%的非小细胞肺癌中失表达或表达减少,而在癌旁正常组织中有80.0%正常表达。WWOX基因的表达与组织类型、细胞分化程度密切相关（P<0.05）,在鳞癌和低分化癌中WWOX基因失表达或表达减少的发生率更高。肺癌中ErbB2的过度表达与肺癌的临床分期、淋巴结转移相关,WWOX蛋白阳性表达与ErbB2负相关(r=–0.239,Ｐ<0.05)。 结论 WWOX蛋白的低表达与ErbB2的过表达可以协同判断预后较差的非小细胞肺癌。     

Expression of the neu gene-encoded protein (P185neu) in human non-small cell carcinomas of the lung

The neu protooncogene is a recently described transforming gene originally isolated from ethylnitrosourea-induced rat neuroblastomas. We have examined the expression of the neu gene in human non-small cell lung carcinomas using immunoprecipitation and immunohistochemistry. The neu protein product (p185neu) was present in eight of 22 non-small cell carcinoma cell lines derived from human lung tumors. Expression of p185neu was found in all histological subtypes of non-small cell carcinomas including large cell carcinomas, squamous cell carcinomas, and adenocarcinomas. Extension of these data to biopsy specimens of human lung tumors demonstrated that normal ciliated bronchial epithelium of the peripheral airways expressed p185neu at low levels. Neoplastic cells in four of 12 adenocarcinomas and three of five squamous cell carcinomas also expressed p185neu at levels higher than the normal ciliated bronchial epithelium. Together these studies indicate that p185neu expression is a common feature of human lung tumors.     

Immunohistochemical study of transforming growth factor-alpha in human lung cancers.

The expression of transforming growth factor-alpha (TGF alpha) was assessed by immunohistochemical staining in 52 human lung tumor samples. All of the 8 small cell lung cancers were negative whereas all of the 18 adenocarcinomas and 23 of the 26 squamous cell carcinomas showed positive immunoreaction to TGF alpha. Distribution of TGF alpha stainings in the squamous cell carcinomas was weaker and more heterogeneous as compared to the adenocarcinomas. Ultrastructural localization of TGF alpha in the squamous cell lung carcinomas by indirect immunogold staining revealed that TGF alpha is present in the cytoplasm as well as the cell membrane but not in the nucleus. This suggests that the lung cancer cells are not only the producer of TGF alpha, but also the target cells of the TGF alpha action. The expression of TGF alpha in lung tumors may be useful diagnostically in differentiating small cell lung cancer from non-small cell lung cancer and may also be important in the study of the biological properties of primary lung cancers.     

Argpyrimidine-modified Heat shock protein 27 in human non-small cell lung cancer: a possible mechanism for evasion of apoptosis

Tumors generally display a high glycolytic rate. One consequence of increased glycolysis is the non-enzymatic glycation of proteins leading to the formation of advanced glycation end-products (AGEs). Therefore, we studied the presence of AGEs in non-small cell lung cancer and consequences thereof. We show the presence of two AGEs, i.e. the major AGE N(epsilon)-(carboxymethyl)lysine (CML) and the methylglyoxal-arginine adduct argpyrimidine, in human non-small cell lung cancer tissues by immunohistochemistry. We found in squamous cell carcinoma and adenocarcinoma tissues a strong CML positivity in both tumour cells and tumour-surrounding stroma. In contrast, argpyrimidine positivity was predominantly found in tumor cells and was strong in squamous cell carcinomas, but only weak in adenocarcinomas (2.6+/-0.5 vs. 1.2+/-0.4, respectively; P<0.005). In accordance, argpyrimidine was found in the human lung squamous carcinoma cell line SW1573, while it was almost absent in the adenocarcinoma cell line H460. Heat shock protein 27 (Hsp27) was identified as a major argpyrimidine-modified protein. In agreement with a previously described anti-apoptotic activity of argpyrimidine-modified Hsp27, the percentage of active caspase-3 positive tumor cells in squamous cell carcinomas was significantly lower when compared to adenocarcinomas. In addition, incubation with cisplatin induced almost no caspase-3 activation in SW1573 cells while a strong activation was seen in H460 cells; which was significantly reduced by incubation with an inhibitor of glyoxalase I, the enzyme that catalyzes the conversion of methylglyoxal. These findings suggest that a high level of argpyrimidine-modified Hsp27 is a mechanism of cancer cells for evasion of apoptosis.     

Novel candidate tumor marker genes for lung adenocarcinoma

Using the representational difference analysis (RDA) technique on pooled mRNA of five primary lung adenocarcinomas and their corresponding non-neoplastic lung tissues, we identified six genes that were putatively overexpressed in this type of lung cancer. Five corresponded to previously isolated genes, while one (Lc19) matched with the sequence of an unannotated EST. Real-time RT-PCR analyses of expression levels in a panel of 34 paired primary non-small cell lung cancer (NSCLC) and corresponding grossly normal appearing lung tissues confirmed the common overexpression of these genes in non-small cell lung cancer. Among these genes, overexpression of Lc19, hyaluronan binding protein 2 (HABP2) and crystalline-mu appeared more specific to adenocarcinoma, whereas ceruloplasmin, integrin alpha-11 and collagen type XI alpha 1 were overexpressed at high frequency among both adenocarcinoma and squamous cell carcinoma. These genes represent novel candidate tumor biomarker genes for NSCLC and its histological subtypes.     

RON及其变异体在肺癌中的表达

目的研究RON及其变异体在肺腺癌中的表达及意义。方法应用免疫组化方法检测106例肺腺癌及35例肺鳞癌中RON的表达情况,并结合临床资料进行统计学分析;通过Western-Blot技术检测31例新鲜肺腺癌组织及9例肺鳞癌组织中RON及其变异体的表达情况,并对RON在不同组织学分级和不同组织学类型中的表达进行统计学分析。结果RON在肺腺癌及鳞癌中都有不同程度的表达,两组比较RON的表达差异具有统计学意义（P〈0.05）;不同分化程度的肺腺癌比较,RON的表达也有差异,低分化的肺腺癌明显高于中分化的腺癌和高分化的腺癌（P〈0.05）;Western-Blot结果显示在肺腺癌和肺鳞癌中均有RON表达;在肺腺癌中观察到RON的变异体,而肺鳞癌中则没有。结论肺癌组织中有RON的过表达,并且肺腺癌中RON的表达明显高于鳞癌（P〈0.05）。肺腺癌中RON的表达与肿瘤分级相关,并存在变异体,而鳞癌则无变异体的表达。     

多聚免疫球蛋白受体(pIgR/SC)在肺癌组织的表达及其临床意义

背景与目的:探讨多聚免疫球蛋白受体(polymeric immunoglobulin receptor/secretory component,pIgR/SC)在肺癌组织中的表达情况及其临床意义.材料与方法:采用大样本石蜡包埋组织(包含344例肺鳞癌、93例肺腺癌、47例小细胞肺癌以及30例同组患者的正常肺组织)构建的组织微阵列,进行免疫组织化学(SP法)染色,分析pIgR/SC蛋白表达情况及其与肺癌临床分期和病理分级间的关系.结果:pIgR/SC蛋白在肿瘤组织(尤其是非小细胞肺癌)中的表达水平(54.1%)显著高于正常对照组织(6.7%),其差异具有统计学意义(P＜0.001);在肺癌组织不同病理类型之间的差异具有统计学意义(P＜0.001),其表达的阳性率从高到低依次为:鳞癌66.3%＞腺癌34.4%＞小细胞肺癌4.3%.而pIgR/SC蛋白表达水平在肺癌的临床分期和组织分级间的差异均无统计学意义(P＞0.05).结论:pIgR/SC蛋白的异常表达反应了肺癌细胞组织学来源的差异,其过表达可能出现在肺癌(尤其是鳞癌)发生发展的早期阶段.     

p185neu expression in human lung adenocarcinomas predicts shortened survival

p185neu is the protein product of the HER2/neu protooncogene. This protein has characteristics of a tyrosine kinase growth factor receptor and is postulated to be important in human carcinogenesis. To define the significance of the expression of this protein in human non-small cell lung cancer, 55 tumors from patients with squamous cell carcinoma (16), adenocarcinoma (29), or large cell carcinoma (10) of the lung were examined for p185neu using immunohistological methods. Five of 16 squamous cell carcinomas and 10 of 29 adenocarcinomas were found to overexpress p185neu relative to levels of expression seen in uninvolved bronchiolar epithelium. For the adenocarcinomas, p185neu expression was associated with older age (66.6 +/- 10.1 versus 57.5 +/- 10.8 years) (P = 0.04) and shortened survival (83.7 +/- 94.1 versus 188.5 +/- 120 weeks) (P = 0.01). In this group, using Cox's multivariate survival analysis, p185neu expression was found to be a significant determinant of survival (P = 0.04) even after accounting for the effect of tumor stage. For the squamous cell carcinomas, p185neu expression was not correlated with any of our clinicopathological parameters. Our findings indicate that non-small cell lung cancers which express p185neu do so at levels higher than that found in normal bronchiolar epithelium, and expression in adenocarcinomas of the lung is independently associated with diminished survival intervals.     

Expression pattern of the scaffold protein IQGAP1 in lung cancer

IQGAP1 is a scaffold protein whose function relates to signal transduction, cell adhesion, local invasion, and distant metastasis of cancer cells. We examined the expression patterns of this protein and clinicopathologic features of lung cancer, and the antibody against IQGAP1 was used for immunohistochemical analysis. Of the 70 surgical specimens examined, there were 40 adenocarcinomas, 19 squamous cell carcinomas, 5 large cell carcinomas, 3 small cell carcinomas, 2 carcinoid tumors, and 1 mucoepidermoid carcinoma. The localization of IQGAP1 was classified into three types: 1) cytoplasmic, 2) membranous, and 3) reduced expression. In adenocarcinoma, the 3 types were observed equally, and differentiation grade was related to the expression pattern. The cytoplasmic type was common in well-differentiated adenocarcinomas, and membranous or reduced expression was frequently seen in moderately- or poorly-differentiated adenocarcinomas. In squamous cell carcinoma, the membranous type was most common. Although the staining pattern of IQGAP1 did not correlate with the positivity of regional lymph nodes, survival in those patients with a cytoplasmic type was significantly better than others with adenocarcinoma (p=0.0144). Expression typing of IQGAP1 in lung cancer was associated with histologic type and can be used to predict survival in patients with adenocarcinoma of the lung.     

Clinical and histopathologic evaluation of the expression of Ha-ras and fes oncogene products in lung cancer.

The expression of Ha-ras and fes oncogenes was investigated with the immunohistochemical method in formalin-fixed, paraffin-embedded tissue specimens of 147 lung carcinomas. Positive immunoperoxidase reactions for Ha-ras p21 were found in 80.5% of the adenocarcinomas, 39.5% of the squamous cell carcinomas, 21.4% of the large cell carcinomas, and 15.4% of the small cell carcinomas; those for fes P85 were found in 51.2% of the adenocarcinomas, 26.3% of the squamous cell carcinomas, 35.7% of the large cell carcinomas, and 15.4% of the small cell carcinomas. Both Ha-ras p21 and fes P85 were expressed most frequently and most strongly in adenocarcinoma. In addition, adenocarcinoma showed significantly higher incidence of concomitant expression of Ha-ras p21 and fes P85 as compared with other histologic types of lung cancer. Thus, the authors suggest that the cooperative effects of Ha-ras and fes oncogenes are especially important in the carcinogenesis of adenocarcinoma. In adenocarcinoma, the incidence and grade of Ha-ras p21 expression increased with the degree of histologic differentiation, suggesting that Ha-ras oncogene might be related to cellular differentiation. Papillary adenocarcinoma showed more frequent Ha-ras p21 expression in comparison with acinar adenocarcinoma. In well- or moderately differentiated adenocarcinoma, the incidence and grade of Ha-ras p21 immunoreactivity in the cases with poor prognosis were significantly higher than in those with good prognosis if other major prognostic factors were equivalent in the two groups. The authors propose that the expression of Ha-ras p21 may be one of the useful prognostic factors in such carcinomas.     

Reduced expression of laminin alpha 3 and alpha 5 chains in non-small cell lung cancers.

The basement membrane is considered to act as a barrier which hinders cancer cells from invading the surrounding stroma. In order to assess changes in essential components during neoplasia in the lung, we immunohistochemically studied distribution patterns of laminins alpha 3 and alpha 5 in 40 adenocarcinomas and 8 squamous cell carcinomas. The a 5 chain was generally preserved at the periphery, frequently disrupted in foci with alveolar collapse and absent in foci of fibroblastic proliferation within adenocarcinomas. Fragmentation and absence of laminin alpha 3 chain were more prominent than for alpha 5 chain. Laminin alpha 3 chain was partially fragmented or absent in peripheral areas of adenocarcinomas, being significantly different from alpha 5 chain. Non-small cell lung cancers with reduced alpha 5 chain showed a tendency for greater lymph node metastasis. In cultured normal air way epithelial cells, both laminin alpha 3 and alpha 5 chains were found to be expressed by northern analysis. Eleven of the twelve cultured lung cancer cell lines did not express alpha 3 chain and expression of alpha 5 chain was reduced in three. Quantitative RT-PCR analysis also demonstrated expression of laminin alpha 3 chain in adenocarcinoma tissues to be significantly lower than in normal lung tissues. These results suggest that expression of laminin alpha chains is often reduced in lung cancer cells and this might contribute to basement membrane fragmentation and subsequent proliferation of stromal elements, as well as play some role in the process of cancer cell invasion.     

ERCC1在非小细胞肺癌中的表达及其与临床病理特征的关系

目的 分析非小细胞肺癌（NSCLC）癌组织中ERCC1的表达状况及其与NSCLC的临床病理特征的关系。方法采用免疫组织化学法检测64例初治中晚期NSCLC患者癌组织中ERCC1基因的表达情况。结果ERCC1表达的阳性率为43．8％，其表达与吸烟有密切关系，在吸烟组中的阳性率（27．8％）显著低于不吸烟组（64．3％）（X^2=8．530，P=0．004）。随着原发灶和疾病分期增加，ERCC1的阳性率有上升的趋势，但组间比较差异均无显著性（均为P〉0．05）。ERCC1在腺癌和低分化癌组中的阳性率分别高于鳞癌和高、中分化组，但差异无显著性（P〉0．05）。ERCC1的表达与年龄、性别和淋巴结转移状况等均未见显著性关联（均为P〉0．05）。结论ERCC1表达与NSCLC的一些临床病理学特征密切相关。     

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。     

Stat3 downstream genes serve as biomarkers in human lung carcinomas and chronic obstructive pulmonary disease

Smoking causes lung cancer and chronic obstructive pulmonary disease (COPD) that impose severe health problem to humans. Both diseases are related to each other and can be induced by chronic inflammation in the lung. To identify the molecular mechanism for lung cancer formation, a CCSP-rtTA/(teto)(7)Stat3C bitransgenic model was generated recently. In this model, persistent activation of the Stat3 signaling pathway induced pulmonary inflammation and adenocarcinoma formation in the lung. A group of Stat3 downstream genes were identified by Affymetrix GeneChip microarray analysis that can be used as biomarkers for lung cancer diagnosis and prognosis. To determine which human lung cancers are related to the Stat3 pathway, multiple Stat3 downstream genes were screened in human lung cancers (adenocarcinomas and squamous cell carcinomas) and lung tissue with COPD. In both cancer and COPD, the Stat3 gene was up-regulated. A panel of Stat3-up-regulated downstream genes in mice was up-regulated in human adenocarcinomas, but not in human squamous cell carcinomas. This panel of genes was also modestly up-regulated in lung tissue with COPD from patients with a history of smoking and not up-regulated in those without histories of smoking. Several Stat3-down-regulated downstream genes also showed differential expression patterns in carcinoma and COPD. These studies support a concept that Stat3 is a potent oncogenic molecule that plays a role in formation of lung adenocarcinomas in both mice and humans. The carcinogenesis of adenocarcinoma and squamous cell carcinoma is mediated by different molecular mechanisms and pathways in vivo. Stat3 and its downstream genes can serve as biomarkers for lung adenocarcinoma and COPD diagnosis and prognosis in mice and humans.