Deletion mapping on the short arm of chromosome 3 in squamous cell carcinoma and adenocarcinoma of the lung.

We examined loss of heterozygosity in 49 adenocarcinomas and 18 squamous cell carcinomas of the lung with 19 RFLP markers on the short arm of chromosome 3. Although no interstitial deletions were observed in any squamous cell carcinomas, interstitial or partial deletions were detected in 23 adenocarcinomas. Identification of two common regions of deletion in adenocarcinomas, at 3p21.3 and 3p14.1-21.1, suggested the presence of at least two tumor suppressor genes on 3p within the same regions commonly deleted in renal cell carcinomas. Correlation between the frequency of loss of heterozygosity on 3p and histopathological grade of adenocarcinoma also was observed. These results imply an etiological difference between two major types of non-small cell lung cancers, adenocarcinoma and squamous cell carcinoma.     

THE STUDY ON RELATIONSHIP BETWEEN CIGARETTE SMOKING AND THE p53 PROTEIN AND P21 PROTEIN EXPRESSION IN NON-SMALL LUNG CANCER

This paper discusses the relationship betweencigarette smoking and the p53 protein and P21 proteinexpression by the immunohistochemical analysis in 93cases with lung cancer in which squamous cell carcinomaaccounted for 45 cases, adenocarcinoma 48 cases. Theresults showed that positive proportion of p53 proteinexpression was 74.20% (28 of 37 squamous cell carcinoma,21 of 30 adenocarcinomas) in cigarette smoking groupwith lung cancers, and 38.46% (3 of 8 squamous cellcarcinoma, 7 of 18 adenocarcinomas) in nonsmokinggroup with lung cancers. The difference was statisticallysignificant. Odds ratio was 4.14 and confidence limitsfor OR was 1.42-12.52. A dose-related presents in thep53 protein expression for the smoking amount andsmoking years. The positive proportion of P21 proteinexpression was 79.31% (21 of 28 squamous cell carcinoma,25 of 30 adenocarcinomas) in cigarette smoking groupwith lung cancers, and 82.75%(10 of 11 squamous,14 of18 adenocarcinomas) in nonsmoking group with lungcancers, the difference     

Overexpression of the aldo-keto reductase family protein AKR1B10 is highly correlated with smokers' non-small cell lung carcinomas.

PURPOSE: Squamous cell carcinoma (SCC) and adenocarcinoma of the lung are currently subject to similar treatment regimens despite distinct differences in histology and epidemiology. The aim of this study is to identify a molecular target with diagnostic and therapeutic values for SCC. EXPERIMENTAL DESIGN: Genes specifically up-regulated in SCC were explored through microarray analysis of 5 SCCs, 5 adenocarcinomas, 10 small cell lung carcinomas, 27 normal tissues, and 40 cancer cell lines. Clinical usefulness of these genes was subsequently examined mainly by immunohistochemical analysis. RESULTS: Seven genes, including aldo-keto reductase family 1, member B10 (AKR1B10), were identified as SCC-specific genes. AKR1B10 was further examined by immunohistochemical analysis of 101 non-small cell lung carcinomas (NSCLC) and its overexpression was observed in 27 of 32 (84.4%) SCCs and 19 of 65 (29.2%) adenocarcinomas. Multiple regression analysis showed that smoking was an independent variable responsible for AKR1B10 overexpression in NSCLCs (P < 0.01) and adenocarcinomas (P < 0.01). AKR1B10 staining was occasionally observed even in squamous metaplasia, a precancerous lesion of SCC. CONCLUSION: AKR1B10 was overexpressed in most cases with SCC, which is closely associated with smoking, and many adenocarcinoma cases of smokers. These results suggest that AKR1B10 is a potential diagnostic marker specific to smokers' NSCLCs and might be involved in tobacco-related carcinogenesis.     

COPD相关性腺癌与鳞癌基因遗传差异的研究进展

在世界范围内,肺癌是癌症死亡的主要原因。其中非小细胞肺癌占85%,主要病理类型为腺癌和鳞癌。研究提示COPD独立于吸烟,为其高危因素。相同的致病因子作用下,COPD相关性腺癌与鳞癌的差异提示,两者之间除常见的慢性炎症、氧化应激、免疫应答等机制外,还存在不同的基因遗传机制。本文就COPD相关性腺癌和COPD相关性鳞癌基因遗传关系的研究进展作一综述。     

肺癌患者血浆外泌体microRNAs标志物的筛选

目的:筛选肺癌患者血浆外泌体中的异常microRNA(miRNA),以期为肺癌筛查及辅助诊断提供新的标志物和方向。方法:选择6名肺癌患者(腺癌3例、鳞癌2例,小细胞肺癌1例)和4名健康对照者血浆样本,采用差速离心法提取血浆外泌体,采用Agilent Bioanalyzer对抽提的RNA进行质量检验,对血浆外泌体miRNA进行高通量测序分析,采用TargetScan、PITA和miRNAorg数据库进行靶基因预测,应用R软件进行靶基因的KEGG和GO分析。结果:miRNA表达分析发现,与正常对照者比较,肺癌患者血浆外泌体中有142种miRNAs表达异常,其中62种miRNAs表达上调,80种miRNAs表达下调。GO分析显示,这些miRNAs的靶基因主要参与了以DNA为模板的转录、转录调控及信号转导等。KEGG分析显示,这些miRNAs的靶基因主要参与轴突导向信号通路、钙信号通路、肌动蛋白细胞骨架调节等。结论:通过对肺癌患者和健康对照者血浆外泌体miRNAs进行高通量测序,初步筛选出差异表达miRNA,经生物信息学分析,推测其对肺癌转移具有良好预测潜力,有望成为肺癌筛查和辅助诊断的候选生物标志物。     

Mutational Damages in Malignant Lung Tumors

Background: Today, genomic changes are an important cause of the occurrence, growth and progression of cancer. Technological advances in cancer genomic analysis platforms have made it possible to identify genomic alterations that may influence response to lung cancer treatment. Methods: The study examined tumor growth-inhibiting oncogenes and genes responsible for cell growth and division to identify mutations characteristic of malignant lung tumors. The mutations were studied in 400 postoperative samples after amplifying p53 and HRAS fragments and p53, p21Waf1, MDM2 mRNA. p53 or p21Waf1 were expressed in 50% of squamous cell carcinomas and adenocarcinomas of the lung. Results: The study examined tumor growth-inhibiting oncogenes and genes responsible for cell growth and division to identify mutations characteristic of malignant lung tumors. The mutations were studied in 400 postoperative samples after amplifying p53 and HRAS fragments and p53, p21Waf1, MDM2 mRNA. p53 or p21Waf1 were expressed in 50% of squamous cell carcinomas and adenocarcinomas of the lung. HRAS mutations were present in most squamous cell carcinomas and adenocarcinomas of the lung. EcoR1- and Pst1- restriction enzymes destroyed the RT-PCR product of the p53 and p21Waf1 mRNA and increased the level of detected mutations in lung adenocarcinoma to 75% and 50 %, respectively. EGFR mutations were more frequent in lung adenocarcinoma than in lung squamous cell carcinoma. Mutations in EGFR exons 19 and 21 found in 65 of 263 lung tumor samples indicated the tumor sensitivity to EGFR tyrosine kinase inhibitors. EGFR deletions in exon 19 occurred mainly in adenocarcinoma, L858R mutations in EGFR exon 21 were quite common in lung adenocarcinoma. Conclusion: The mutations detected in most squamous cell carcinomas and adenocarcinomas of the lung could be used to diagnose and predict the disease severity and targeted therapy efficacy.     

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.     

癌基因K-ras、C-myc、erbB-2的激活与非小细胞肺癌组织学类型相关性研究

为探讨癌基因突变、扩增与非小细胞肺癌发生及类型间的关系，应用核酸杂交的方法检测了63伤1肺手术标本中（腺癌19例，鳞癌23例，腺鳞癌3例，大细胞癌3例，小细胞癌3例，其它12例）三种癌基因（c－myc、K－ras、erbB－2）的扩增及突变。实验结果显示：50％（24／48）的非小细胞癌分别出现三种癌基因的激活；31．6％（6／19）的腺癌出现K－ras12密码子突变及K－ras扩增；30．4％（7／23）的鳞癌有erbB－2扩增；10．4％（5／48）的非小细胞癌存在c－myc扩增。提示K－ras突变可能为肺腺癌所特有，而erbB－2扩增是肺鳞癌发生的重要因素。     

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.     

Cell type specificity of lung cancer associated with arsenic ingestion.

Arsenic is a well-documented human carcinogen. Previous studies on urinary bladder and skin cancers have shown that arsenic can cause specific cell types of malignancy. To evaluate whether this is also true for lung cancers, we conducted a study on 243 townships in Taiwan. We identified patients through the National Cancer Registry Program and compared the proportion of each major cell type between an endemic area of arsenic intoxication with exposures through drinking water, which includes 5 of the townships and the other 238 townships. To control for gender and age, we analyzed data on men and women separately and divided patients into four age groups. A total of 37,290 lung cancer patients, including 26,850 men and 10,440 women, was diagnosed between January 1, 1980 and December 31, 1999 in study townships. Patients from the endemic area had higher proportions of squamous cell and small cell carcinomas, but a lower proportion of adenocarcinomas. These findings were similar across all age groups in both genders, although the lack of data on smoking is a limitation of our study. The results suggested that the carcinogenicity of arsenic on lungs is also cell type-specific: squamous cell and small cell carcinomas appeared to be related to arsenic ingestion, but not adenocarcinoma. Whereas data in the literature are limited, the association between adenocarcinoma and arsenic exposures through inhalation appeared to be stronger than that of squamous cell carcinoma. Therefore, we speculate that arsenic may give rise to different mechanisms in the development of lung cancers through different exposure routes.     

Molecular heterogeneity of non‐small cell lung carcinoma patient‐derived xenografts closely reflect their primary tumors

Availability of lung cancer models that closely mimic human tumors remains a significant gap in cancer research, as tumor cell lines and mouse models may not recapitulate the spectrum of lung cancer heterogeneity seen in patients. We aimed to establish a patient‐derived tumor xenograft (PDX) resource from surgically resected non‐small cell lung cancer (NSCLC). Fresh tumor tissue from surgical resection was implanted and grown in the subcutaneous pocket of non‐obese severe combined immune deficient (NOD SCID) gamma mice. Subsequent passages were in NOD SCID mice. A subset of matched patient and PDX tumors and non‐neoplastic lung tissues were profiled by whole exome sequencing, single nucleotide polymorphism (SNP) and methylation arrays, and phosphotyrosine (pY)‐proteome by mass spectrometry. The data were compared to published NSCLC datasets of NSCLC primary and cell lines. 127 stable PDXs were established from 441 lung carcinomas representing all major histological subtypes: 52 adenocarcinomas, 62 squamous cell carcinomas, one adeno‐squamous carcinoma, five sarcomatoid carcinomas, five large cell neuroendocrine carcinomas, and two small cell lung cancers. Somatic mutations, gene copy number and expression profiles, and pY‐proteome landscape of 36 PDXs showed greater similarity with patient tumors than with established cell lines. Novel somatic mutations on cancer associated genes were identified but only in PDXs, likely due to selective clonal growth in the PDXs that allows detection of these low allelic frequency mutations. The results provide the strongest evidence yet that PDXs established from lung cancers closely mimic the characteristics of patient primary tumors.     

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。     

Detection of ras gene mutations in human lung cancers by single-strand conformation polymorphism analysis of polymerase chain reaction products

A simple, sensitive method of DNA analysis of nucleotide substitutions, namely, single-strand conformation polymorphism analysis of polymerase chain reaction products (PCR-SSCP analysis), was used for detection of mutated ras genes in surgical specimens of human lung cancer. Of a total of 129 tumors analysed, 22 contained a mutated ras gene. Of the 66 adenocarcinomas analysed, 14 contained an activated c-Ki-ras2 gene (the mutations in codon 12 in 6, in codon 13 in 4, in codon 18 in one, and in codon 61 in 3), one contained a c-Ha-ras1 gene with a mutation in codon 61 and 3 contained N-ras genes with mutations (in codon 12 in one and in codon 61 in 2). Mutated rats genes were also found in 2 of 36 squamous cell carcinomas (c-Ha-ras1 genes with mutations in codon 61) and 2 of 14 large cell carcinomas (c-Ki-ras2 genes with mutations in codon 12). No mutation of the ras gene was detected in 8 small cell carcinomas and 5 adenosquamous cell carcinomas. These results indicate that activation of the ras gene was not frequent (17%) in human lung cancers, that among these lung cancers mutation of the ras gene was most frequent in adenocarcinomas (27%) and 73% of the point mutations were in the c-Ki-ras2 gene in codon 12, 13, 18 or 61.     

黑色素瘤抗原基因MAGE-1、MAGE-3和抑癌基因p53在肺癌中表达的研究

目的:分析肺癌组织MAGE-1和MAGE-3基因以及p53基因表达及其相互关系。方法:取肺癌组织标本30例,另取癌周组织作为对照。采用逆转录聚合酶链反应(RT-PCR)方法,检测MAGE-1和MAGE-3基因以及p53基因的表达。结果:MAGE-1,MAGE-3在肺癌中有较高表达,MAGE-3的表达高于MAGE-1的表达。p53在肺癌中表达较低,在正常组织中表达较高;MAGE-1在肺腺癌中表达较鳞癌高,MAGE-3在肺鳞癌中表达较高;MAGE-1,MAGE-3的表达可能与肿瘤分化程度无关;p53在小细胞肺癌中的表达有待进一步研究;p53与MAGE在体内表达成负相关。结论:MAGE-1、MAGE-3和p53在肺癌组织与正常组织中有不同的表达,MAGE-1、MAGE-3的表达与病理类型有关,与肿瘤分化无关。     

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.     

Glycosaminoglycans in human lung cancer

The quantitative changes of glycosaminoglycans in tumor tissue of human lung cancers (2 squamous cell carcinomas, 4 adenocarcinomas and 5 small cell carcinomas) were studied. The total amount of glycosaminoglycans in human lung cancer tissues increased 1.4 to 4 times in comparison with that in normal lung tissues. The increase in tissue content of glycosaminoglycans was accompanied by an increase in the chondroitin sulfate level in every histologic type of lung cancer, as well as by a marked increase in hyaluronic acid level in squamous cell carcinomas, and a moderate increase in its level in small cell carcinomas. The concentrations of dermatan sulfate and heparan sulfate in lung cancer tissues did not show any significant changes compared with those in normal lung tissues. The increase in total amount and changes in the composition of glycosaminoglycans in human lung cancer tissue were closely related to the histologic type of the tumor.     

Clinicopathologic characteristics of adenosquamous carcinoma of the lung

Fifty-six cases of surgically resected adenosquamous carcinoma of the lung were studied clinicopathologically, and their outcome was compared with that of adenocarcinomas and squamous cell carcinomas of the lung. The frequency rate of adenosquamous carcinoma was 2.6% of 2160 primary lung cancers resected in the National Cancer Center Hospital (Tokyo, Japan). The survival curves of patients with adenosquamous carcinomas, adenocarcinomas, and squamous cell carcinomas indicated that the outcome of adenosquamous carcinoma was poorer than that of adenocarcinomas and squamous cell carcinomas, particularly in Stages I and II. The amount of adenocarcinoma component did not affect the survival rate, although the histologic features of metastatic lymph nodes was somewhat influenced by the histologic type of the primary tumors. The histologic subtype of adenosquamous carcinoma was one of the independent prognostic determinants.