Normal Bronchial Tissue Research Articles

Loss of cables protein expression in human non–small cell lung cancer: A tissue microarray study

Loss of heterozygosity (LOH) on chromosome 18q is common in lung cancer. The genes involved in LOH on 18q in lung cancer have not been well characterized. Cables, a cyclin-dependent kinase (cdk) interacting protein, has recently been identified and mapped to human chromosome 18q11-12. Cables inhibits cell growth and suppresses tumor formation in nude mice, making it a candidate gene for 18q LOH in lung cancer. Little is known regarding Cables protein expression in human non–small cell lung cancer (NSCLC). In this study we examined Cables expression in 163 NSCLC and nonneoplastic lung specimens using tissue microarrays. Strong nuclear staining was present in normal lung and bronchial tissue. We also evaluated the Cables protein expression pattern and its correlation with histopathologic features and with clinical course of NSCLC. The results of the present study demonstrate for the first time that numerous NSCLCs (45%) lose Cables expression. Furthermore, more adenocarcinomas show a loss of this novel protein than do squamous counterparts. The relationship between tumor histology type and Cables expression appears to be statistically significant (P = 0.028). Our results suggest that Cables may be involved in the pathogenesis of NSCLC. HUM PATHOL 34:143-149. Copyright 2003, Elsevier Science (USA). All rights reserved.

OCT (Optical Coherence Tomography) for Diagnosis of Bronchial Lesions: Preclinical Experience

PURPOSE: OCT can obtain high-resolution, cross-sectional microimages of tissue potentially allowing for optical biopsy to substitute for conventional excisional biopsy. The purpose of the present study is to investigate the capability of OCT to image the microstructure of the normal and abnormal bronchial tissue.

How to combine cytotoxic and cytostatic agents

In the last two decades some progress has been made in the medical treatment of advanced non-small cell lung cancer (NSCLC) with improvement of polychemotherapy outcomes. Nonetheless, optimization of polychemotherapeutic regimens will not likely produce a substantial increase of patient survival; palliation and quality of life of patients remain the first goal of therapy. As a consequence, investigating the association of new biologic agents with chemotherapy is mandatory, even if the search for the best polychemotherapeutic regimen is still ongoing. Two very large recent randomized clinical trials, performed by the Southwest Oncology Group (SWOG) and the Eastern Cooperative Oncology Group (ECOG) [1,2] gave very significant answers on this issue, showing that the most commonly used new platinum-based regimens are similarly effective in terms of survival, differing only in their toxicity profile. Fortunately, our improved knowledge of tumor biology and mechanisms of oncogenesis suggests several new potential targets for clinical research in cancer therapy. Among the most promising fields of research in this direction there are the epidermal growth factor receptor (EGFR) and attempts to block its activity [3], and cyclooxigenase inhibitor (anti-COX 2) [4,5]. IRESSA is one the most exciting agent acting on EGFR with a favorable toxicity profile and available as oral formulation. Recently, in the Multicenter Italian Lung cancer in the Elderly Study (MILES) we showed that combination chemotherapy with gemcitabine /vinorelbine does not improve any outcome as compared with single agent chemotherapy with gemcitabine or vinorelbine in advanced NSCLC elderly patients [6]. In the same patients population we started a phase II randomized trial of gemcitabine /IRESSA and vinorelbine / IRESSA. Primary end point of the trial is to evaluate activity and toxicity of these new combinations. Adding a promising new biologic agent to single agent chemotherapy could improve current results obtained in this particular subgroup of patients. Cyclooxigenase (COX) catalyses the synthesis of prostaglandins (PGs) from arachidonic acid. There are at least two isoforms of COX: COX-1, which is constitutively expressed in many tissues, and COX-2, not constitutively expressed but frequently induced by various stimuli, such as mitogens, cytokines, carcinogens and growth factors. A substantial body of evidence indicates that COX-2 and PGs play an important role in tumorigenesis. Mechanisms involved in COX-2 participation in tumor growth include angiogenesis stimulation, the inhibition of immune surveillance and the inhibition of apoptosis. COX-2 is overexpressed in various human malignancies and represents an important early event in the development of some human tumors, including lung cancer. Precursor lung cancer lesions such as atypical alveolar epithelium, atypical adenomatous hyperplasias and carcinomas in situ frequently exhibit COX-2 overexpression relative to surrounding normal bronchial tissue. COX-2 upregulation has also been found in up to 90% of invasive lung tumors and is particularly common in well-differentiated adenocarcinomas and, to a lesser extent, in the squamous subtype. Adenocarcinoma cells metastatic to mediastinal lymph nodes also express COX-2 at a higher proportion and level than corresponding primary tumors, suggesting that COX-2 overexpression may enhance metastatic potential; moreover, COX-2 overexpression is a marker of poor prognosis in NSCLC. In various preclinical studies, COX-2 inhibition was shown to produce inhibition of tumor growth, both in vitro and in vivo. Preclinical studies in vivo showed that a selective COX-2 inhibitor, rofecoxib, also has potential as a chemopreventive agent in human intestinal and colon cancer. These evidences support the hypothesis that selective COX-2 inhibitors may prove beneficial in the prevention and treatment of many human tumors, including lung cancer. Based on this background we launched a phase III randomized trial named GEmcitabine-Coxib (GECO), evaluating the E-mail address: cgridelli@sirio-oncology.it (C. Gridelli). Lung Cancer 38 (2002) S39 /S40

HER-2/neu overexpression in patients with radically resected nonsmall cell lung carcinoma. Impact on long-term survival.

Using immunohistochemistry, the authors prospectively investigated the expression of HER-2/neu protein in radically resected specimens of nonsmall cell lung carcinoma (NSCLC) and evaluated its impact on long-term prognosis. Between January 1991 and February 1992, surgical specimens from 130 consecutive patients who underwent radical resection for NSCLC (60 squamous cell carcinoma, 48 adenocarcinoma cases, and 22 large cell carcinomas) and that were staged (according to the TNM staging system) pathologically as Stage I (41 cases [32%]), Stage II (37 cases [28%]), and Stage IIIA (52 cases [40%]) were investigated for the expression of HER-2/neu using an avidin-biotin complex immunohistochemical technique. A semiquantitative four-stage grading system was used (0%, 1-5%, 6-20%, and > 20% positive cells) and an average number of 1500 cells/section was considered. Data were correlated with clinical and pathologic variables. Normal bronchial tissue was found to be completely negative for HER-2/neu expression whereas 21 of the 130 tumor specimens (16%) were positive (range 1-> 20%). HER-2/neu positivity did not appear to differ significantly among pathologic stages and histotypes. Using a predetermined cutoff value of 5% positive cells, 15 tumor specimens (12%) were found to be above this value. The median survival time (85 weeks vs. 179 weeks) and overall survival rate were significantly lower in patients with > 5% HER-2/neu-positive tumors (hazard ratio for the group with > 5% positive cells: 2.94, 95% confidence interval, 1.62-5.34; P < 0.0004). On multivariate analysis, HER-2/neu and extent of tumor emerged as independent factors for disease-related mortality. In NSCLC, the negative impact of HER-2/neu overexpression on survival was maintained in the long-term follow-up of radically resected patients. HER-2/neu overexpression may be a valuable prognostic factor as well as a potential target for biologic therapies.

Spectroscopic analysis of the autofluorescence from human bronchus using an ultraviolet laser diode.

A GaN based ultraviolet (UV) laser diode (LD) was used to study the autofluorescence (AF) spectrum of the normal and tumor human bronchial tissues under ex vivo conditions. The UV LD generates a coherent short wavelength (around 400 nm) light beam with an intensity of about a few watts. AF spectrum data can be obtained without interference by excitation light. A clear blue peak located at around 483 nm was observed along with a green peak at around 560 nm in the normal tissue. The peak intensities observed were very weak for the tumor tissues. The AF imaging and spectrum analysis were performed along with a histopathological study. The spatial distribution of the elastin in the bronchial tissue affected the intensity of the AF whereas the spectrum shape was not affected. Strong AF was observed from regions that include a high density of the elastin. Biopsy measurements were performed for ex vivo samples, and depth profiling of the elastin was studied along with variations of the AF spectrum. AF spectra excited by the UV LD for fluorescence materials including FAD, NADH, and elastin were measured. The spectrum shape of the elastin as well as of NADH was similar to that of normal bronchial tissues.

Spectroscopic differentiation between normal bronchial and lung cancer tissues by laser-induced fluorescence (LIF) with intravenous injection of hematoporphyrin monomethyl ether (HMME)

To study the feasibility of laser-induced fluorescence ( LIF) spectroscopy with intravenous injection of a new agent, hematoporphyrin monomethyl ether (HMME) , in the diagnosis of lung cancer. Fifteen patients with lung cancer were administrated HMME 2. 5 mg/ kg intravenously three hours before lobectomy. Surgical specimens were collected for examination of LIF spectrum. TheLIF spectra of normal bronchial and lung cancer tissues were measured with a detecting system which consists of an YAG laser ( wavelength 355 nm) and an optical multichannel analyzer ( OMA) . The patholog ical examination was performed in each specimen. The fluorescence intensity of the lung cancer tissues ( 31 446±5 017) was much lower than that of the normal bronchial tissues ( 75 430±8 908) ( P < 0. 001) . There was a flat spectrum in the normal bronchial tissues at the wavelength from 580 to 600 nm ( I580nm / I600nm = 1. 081 ±0. 090) , but the cancer tissue spectrum showed a smoothly descending profile ( I580nm/ I600nm = 1. 260±0. 157) . A remarkable characteristic peak ( drug peak) located at the 623. 4 nm±1. 6 nm in the lung cancer spectrum. Using the criterion of I580nm/ I600nm , the sensitivity, specificity, and accuracy of the diagnosis for lung cancer were 80. 0%, 73. 9%, and 76. 7%, respectively , and were 95. 0%, 91. 3% and 93. 0% respectively according to the criterion of the slope of the drug peak. The LIF spectroscopy following intravenous injection of the new photosensitiser HMME can differentiate lung cancer from normal bronchus and can increase the sensitivity, specificity and the accuracy comparing to the laser-induced auto fluorescence in the diagnosis of lung cancer.

Bronchial mucosa produced by tissue engineering: A new tool to study cellular interactions in asthma

Background: The use of fiberoptic bronchial biopsies has improved our understanding of the immunopathology of asthma. However, this approach offers a limited ability to perform mechanistic studies observing cell-cell and cell-matrix interactions, which are a key issue in the study of airway remodeling. Tissue engineering is a technique that combines the use of biology and engineering expertise to generate a limitless amount of tissue from small samples. This technology allows for the study of cell interactions under conditions as close as possible to the natural environment. Objective: The aim of this study was to evaluate the feasibility of an engineered human bronchial mucosa as a model to study cellular interactions in asthma. Methods: Human bronchial fibroblasts from normal and asthmatic donors were incorporated into collagen gel. Bronchial epithelial cells were seeded over this gel and then cultured in an air-liquid interface in the presence or the absence of T lymphocytes. Biopsy specimens from these engineered mucosa were taken for structural and ultrastructural analysis, and T lymphocytes were harvested and used to localize IL-5. Results: Histologic analysis showed that engineered mucosa with normal bronchial cells presented a pseudostratified ciliated epithelium with the presence of mucus secretory cells. The electron microscopy analysis confirmed these histologic results. These features were comparable with those observed in normal bronchial tissues. However, in engineered mucosa from asthmatic subjects, the tissue structure was disorganized, particularly the epithelial cell arrangement. The percentage of IL-5+ lymphocytes was significantly (P = .03) higher in engineered bronchial mucosa from asthmatic subjects (87% ± 2%) compared with mucosa from normal volunteers (2% ± 0.3%). Conclusion: Using tissue engineering, we produced an in vitro model of bronchial mucosa from normal and asthmatic subjects. These models could be a valuable tool to better understand key mechanisms involved in inflammation and airway repair. (J Allergy Clin Immunol 2001;107:36-40.)

Early Detection of Lung Cancer With Laser-Induced Fluorescence Endoscopy and Spectrofluorometry

We performed a clinical trial of laser-induced fluorescence endoscopy (LIFE) for detection of precancerous lesions and cancer including carcinoma in situ (CIS), which are difficult to detect by white-light bronchoscopy. Results with LIFE were compared with the criterion standard, white-light bronchoscopy. The evaluation of these endoscopic results spectrofluorometrically was examined, and pixels of LIFE images composed of digital signals for the intensities of red and green were analyzed. Tertiary-level hospital treating referrals and subjects with suspicious results in mass screening. We examined 65 subjects with suspected lung cancer by both methods, and performed biopsy on 216 lesions. The accuracy of diagnosis by white-light bronchoscopy, with histopathologic results as the standard, was 48.6%. The accuracy by LIFE was 72.7%. The sensitivity of conventional bronchoscopy for detection of severe dysplasia (21 biopsy specimens) or cancer (28 biopsy specimens) was 61.2% and specificity was 85.0%. With results by LIFE added, these values were 89.8% and 78.4%, respectively. Of nine patients with CIS, only LIFE showed one lesion, and only LIFE showed the extent of seven of the lesions. The autofluorescence of eight lesions was measured spectrofluorometrically; normal bronchial tissue, severe dysplasia, and cancerous tissue had spectral differences. The red/green intensity of cancers on histograms of LIFE images generally was greater than the ratios for metaplasia or normal bronchial wall. Use of both methods should facilitate early detection. Evaluation by spectrofluorometry and analysis of digital signal intensity of results by LIFE make results more objective.

Analyses of bronchial bulky DNA adduct levels and CYP2C9, GSTP1 and NQO1 genotypes in a Hungarian study population with pulmonary diseases.

Carcinogen-DNA adducts may represent an intermediate end-point in the carcinogenic cascade and may reflect exposure to chemical carcinogens, as well as susceptibility and, ultimately, cancer risk. Interindividual variability in activity of enzymes involved in the metabolism of polycyclic aromatic hydrocarbons to mutagenic diol epoxides may predict adduct levels and, indirectly, lung cancer risk. Using 32P-postlabeling methods, the levels of bulky DNA adducts were determined in macroscopically normal bronchial tissues obtained from resected lobes of 143 Hungarian patients with lung malignancy and other pulmonary conditions. DNA from normal tissue was also evaluated for polymorphisms in cytochrome P450 2C9 (CYP2C9) at two sites, codons 144 (Arg/Cys) and 359 (Ile/Leu), for glutathione S-transferase P1 (GSTP1) at codon 105 and for NAD(P)H:quinone oxidoreductase (NQO1) at codon 187 (Pro/Ser). Using the Mann-Whitney U-test and analysis of variance, levels of adducts were evaluated in relation to variant genotypes, separately for smokers and non-smokers. As previously reported, bulky DNA adduct levels in smokers (n = 104) were estimated to be 54% higher than in non-smokers (n = 39) (8.6 +/- 4.2 versus 5.6 +/- 3.3 per 10(8) nucleotides, respectively, P < 0.01). Adduct levels were 16-29% higher in individuals with the homozygous Ile359/Ile359 CYP2C9 allele than in those heterozygous for the variant allele (Ile359/Leu359) [8.8 +/- 4.3 (n = 84) versus 7.6 +/- 3.5 (n = 20) for smokers and 5.8 +/- 3.5 (n = 32) versus 4.5 +/- 1.3 (n = 7) for non-smokers], although differences were not statistically significant. There were no clear differences in adduct levels in relation to genotypes of NQO1 or GSTP1. Although numbers of patients in this study are large in relation to many studies of carcinogen-DNA adducts, it is still possible that significant differences were not noted for polymorphisms in xenobiotic metabolizing enzymes due to relatively small numbers in stratified data.

Isolation of protein kinase C-alpha-regulated cDNAs associated with breast tumor aggressiveness by differential mRNA display.

Elevated levels of protein kinase C (PKC) are associated with increased metastatic capacity in both human breast cancer cells and breast tumors. MCF-7 breast cancer cells stably transfected with PKC-alpha were recently shown to display a more aggressive phenotype and increased tumorigenicity in nude mice. To identify genes involved in the progression to the aggressive phenotype, mRNA differential display was performed to isolate cDNAs that are differentially expressed between the parental, non-metastatic MCF-7 cell line and the metastatic derivative MCF-7-PKC-alpha cell line. One cDNA was identified which was upregulated and four cDNAs were downregulated in MCF-7-PKC-alpha cells. The upregulated cDNA may be a differentiation-specific gene as it is 100% homologous to a putative glialblastoma cell differentiation-related protein, GBDR1. DNA sequence analysis and flow cytometry revealed that three of the downregulated cDNAs correspond to histone 3.B, and integrins alpha3 and alpha6. The fourth downregulated cDNA clone, G2Q, is a novel sequence. G2Q is expressed in normal breast and bronchial tissue, but is downregulated in a variety of tumor cell lines and in aggressive primary and secondary breast tumors, suggesting that G2Q may be a useful prognostic indicator of tumor aggressiveness. Further, downregulation of G2Q expression in the non-metastatic MCF-7 cells by antisense oligonucleotides resulted in increased in vitro invasive capacity of these cells in a Matrigel matrice. This study provides the basis for identifying new genes involved in breast tumor progression and the role that PKC plays in the pathogenesis of this cancer.

B7 costimulation is required for IL-5 and IL-13 secretion by bronchial biopsy tissue of atopic asthmatic subjects in response to allergen stimulation.

Asthma is a complex disorder characterized by airway hyperreactivity and inflammation. To analyze cellular interactions required for the secretion of cytokines by the bronchial mucosa, we have evaluated the ex vivo response of tissue explants to allergen. Endobronchial mucosal biopsy tissue from mild atopic asthmatic subjects and normal control subjects were maintained in culture for 24 h. To detect reactivity to allergen, the explants were stimulated with dust mite extract Dermatophagoides pteronyssinus (Der p). Our analysis revealed that without any overt stimulation, mRNA transcripts for interleukin (IL)-5 and IL-13 were expressed by asthmatic but not normal bronchial tissue. In contrast, the expression of interferon-gamma was observed in a higher proportion of cultured bronchial biopsies from the normal control subjects than in those from asthmatic subjects. Addition of Der p allergen did not change the cytokine profile of the explants from control volunteers but augmented the expression of IL-5 mRNA and induced secretion of the protein by the asthmatic bronchial tissue. In most cases, allergen also increased the production of IL-13 by bronchial tissue from asthmatic subjects. The allergen-induced secretion of IL-5 and IL-13 was inhibited by the fusion protein CTLA-4Ig, reflecting a requirement for CD80 (B7-1) and/or CD86 (B7-2) costimulation for the expression of the Th2 cytokines. This requirement for B7/CD28 costimulation is consistent with the hypothesis that IL-5 and IL-13 are produced by allergen-specific T cells resident in the asthmatic bronchial mucosa.

P53 and c-erbB2 as the Immunohistochemical Markers in Patients with Sguamous Cell Carcinoma of the Lung

Background: With the development of the molecular biological methods, studies of the early diagnosis of lung cancer and the detection in the preneoplastic state by using genetic probes in the high risk groups are widely investigated. In lung cancer, squamous cell carcinoma is considered to progress from the normal bronchial mucosa to the preneoplastic state, and finally to the invasive carcinoma. In this study, we investigated the expression of p53 and c-erbB2 in the normal bronchi and the cancer tissues in patients with squamous cell lung cancer to evaluate the possibility of using these immunohistochemical markers as the diagnostic and prognostic parameters of patients with squamous cell lung cancer. Method: The normal and cancerous bronchial tissues of 25 patients with squamous cell carcinoma of the lung, surgically resected from May 1995 to November 1996, were immunohistochemically stained with the monoclonal antibodies to p53(DAKO-p53) and c-erbB2(phamingen 15821A) respectively. We compared the expression status of these markers between the normal bronchial mucosa and the tumor tissue, and also investigated the relationship between the expression status of these markers in tumor tissues and the pathological stage, and the survival time. Results: The pathological stage was as follows; stage I, II were found in 5 patients respectively, stage IIIA was in 8 patients, stage IIIB was in 4 patients, and stage IV was in 3 patients. The expression rate of p53 in the squamous cell lung cancer was 48%, and it was not expressed in the normal bronchial mucosa. The expression status was increased as the pathological stage advanced(p=0.0091 by test of trend). But there were no relationship between the expression of p53 and the median survival time. C-erbB2 did not yield a significantly meaningful result. Conclusion: p53 was not found in the normal bronchial mucosa, but it was expressed in 48% of the tumor tissue. And the expression rate increased as the pathological stage advanced. So it would be helpful to apply the immunochistochemical stain with p53 in the bronchial biopsy specimen in the early diagnosis trial or staging of squamous cell lung cancer.

Smoking-associated bulky DNA adducts in bronchial tissue related to CYP1A1 MspI and GSTM1 genotypes in lung patients.

Relationships between smoking status and levels of bulky DNA adducts were investigated in bronchial tissue of lung patients in relation to their GSTM1 and CYP1A1 MspI genotypes. A total of 150 Hungarian patients undergoing pulmonary surgery were included in the study, 124 with lung malignancies and 26 with non-malignant lung conditions. There were significant relationships between smoking status and bulky DNA adduct levels, as determined by 32P-post-labelling analysis, in macroscopically normal bronchial tissues. There was a highly significant difference in the adduct levels of a combined group consisting of current smokers and short-term ex-smokers (< or = 1 year abstinence) compared with life-time non-smokers and long-term ex-smokers (> 1 year abstinence) (P = 0.0001). The apparent half-life was estimated to be 1.7 years for bulky DNA adducts in the bronchial tissue from ex-smokers. There were no statistically significant correlations between (i) daily cigarette dose and DNA adduct levels in current smokers, (ii) DNA adduct level and histological type of lung cancer, or (iii) GSTM1 and CYP1A1 MspI genotypes and DNA adduct levels after adjustment for either smoking status or malignancy. By multiple logistic regression analysis, smoking and GSTM1 null genotype were found to be risk factors for squamous cell carcinoma. However, bulky DNA adduct levels in bronchial tissue did not appear to be a statistically-significant risk factor for the major histological types of lung cancer.

MDR1-Pgp 170 expression in human bronchus.

MDR1 P-glycoprotein (Pgp 170), a member of the adenosine triphosphate (ATP) binding cassette transporters, acts as an efflux pump for various hydrophobic agents, particularly for xenobiotics such as benzo(a)pyrene. It has also been shown to regulate cell-volume activated chloride channels. Pgp 170 could, therefore, be of particular importance in cellular mechanisms of defence in the airways and in the control of mucus layer composition. For these reasons, we evaluated the precise localization of Pgp 170 in human adult airways. Fresh non-neoplastic bronchial specimens were collected from 33 patients (26 smokers, four exsmokers and three nonsmokers) who underwent surgery for lung carcinoma. The presence of MDR1 messenger ribonucleic acid (mRNA) was demonstrated by reverse transcriptase chain reaction (RT-PCR) in bronchial epithelial cells collected by gentle scraping from either smokers, exsmokers or nonsmokers. Immunodetection of Pgp 170 using a panel of monoclonal antibodies (MRK16, JSB1, C219, C494) was performed either on cryostat or paraffin-embedded sections of histologically normal bronchial tissue. Pgp 170 was constantly detected with intense labelling at the apical surface of ciliated epithelial cells from the surface epithelium or ciliated collecting ducts, and on apical and lateral surfaces of serous cells from bronchial glands. No staining of mucus-secreting cells was observed. Pgp 170 was also demonstrated at the luminal surface of endothelial cells of bronchial capillaries. In conclusion, the expression of MDR1 P-glycoprotein in bronchial structures, particularly at the epithelial apical surface, suggests important roles for this transmembrane protein in human airways.

Suppression of Retinoic Acid Receptor in Non-Small-Cell Lung Cancer In Vivo: Implications for Lung Cancer Development

Retinoids, analogues of vitamin A, are required for the normal growth and differentiation of human bronchial epithelium. They are also able to reverse premalignant lesions and prevent second primary tumors in some patients with non-small-cell lung cancer (NSCLC). These effects are thought to result from modulation of cell growth, differentiation, or apoptosis (programmed cell death). When certain retinoid receptors in the cell nucleus (i.e., retinoic acid receptors [RARs] and retinoid X receptors [RXRs]), which mediate most retinoid actions, are suppressed, abnormal activity may result that could enhance cancer development. This study was designed to determine whether there are abnormalities in the expression of retinoid receptors in surgical specimens from patients with NSCLC. Transcripts of nuclear retinoid receptors were detected in formalin-fixed, paraffin-embedded specimens by use of digoxigenin-labeled riboprobes specific for RAR alpha, RAR beta, RAR gamma, RXR alpha, RXR beta, and RXR gamma for in situ hybridization to histologic specimens from 79 patients with NSCLC and as control from 17 patients with non-lung cancer. The quality and specificity of the digoxigenin-labeled probes were determined by northern blotting, and the specificity of the binding of antisense riboprobes was verified by use of sense probes as controls. All receptors were expressed in at least 89% of control normal bronchial tissue specimens from 17 patients without a primary lung cancer and in distant normal bronchus specimens from patients with NSCLC. RAR alpha, RXR alpha, and RXR gamma were expressed in more than 95% of the NSCLC specimens. In contrast, RAR beta, RAR gamma, and RXR beta expression was detected in only 42%, 72%, and 76% of NSCLC, respectively. These data suggest that the expression of RAR alpha, RXR alpha, and RXR gamma is not altered in NSCLC; however, expression of RAR beta and possibly also of RAR gamma and RXR beta is suppressed in a large percentage of patients with lung cancer. The loss of expression of one or more of these nuclear retinoid receptors may be associated with lung carcinogenesis.

Eosinophils as a potential source of platelet-derived growth factor B-chain (PDGF-B) in nasal polyposis and bronchial asthma.

Tissue remodeling in bronchial tissues from asthmatics as well as in nasal polyp (NP) tissues includes sub-basement membrane deposition of collagen, stromal deposition of extracellular matrix protein, and hypertrophy/hyperplasia of airway smooth muscle cells, which are relevant to the cellular and molecular events induced by platelet-derived growth factor (PDGF). Therefore, we investigated the localization of mRNA and protein of PDGF-B chain (PDGF-B) in NP tissues and bronchial tissues from mild and severe asthmatics by in situ hybridization and immunohistochemistry, respectively. Cells expressing PDGF-B mRNA were found in all nine NP tissues and in bronchial tissues from 2 of 6 normal subjects, 2 of 5 mild asthmatics, and all of 6 severe asthmatics examined. The vast majority of cells expressing PDGF-B mRNA were eosinophils in NP (99.7 +/- 0.2%, mean +/- SD) and asthmatic bronchial tissues (75.0 and 77.8% in mild asthma, and 92.7 +/- 8.1% in severe asthma), but no cells expressing PDGF-B mRNA were eosinophils in normal bronchial tissues. The number of cells expressing the gene in severe asthma tissues (122.3 +/- 32.2/mm2) was similar to that in NP tissues (152.8 +/- 73.9/mm2) and greater than that in mild asthma tissues (4.7 +/- 7.6/mm2, P < 0.01), which was not significantly greater than that in normal bronchial tissues (3.4 +/- 5.2/mm2). Furthermore, we detected immunolocalization of PDGF-B in NP tissues and in asthmatic bronchial tissues. The eosinophils purified from peripheral blood were demonstrated to express PDGF-B gene transcript and immunoreactivity after stimulation with A23187.(ABSTRACT TRUNCATED AT 250 WORDS)

Laser-induced fluorescence spectroscopy at endoscopy: tissue optics, Monte Carlo modeling, and in vivo measurements

The optical properties (absorption coefficient, scattering coefficient and the anisotropic factor of scattering) and fluorescence characteristics of normal and abnormal bronchial tissue were measured in vitro. After adding additional blood optical properties to in vitro optical properties of tissue, the <i>in vivo</i> bronchial fluorescence was simulated and analyzed by Monte Carlo modeling. The Monte Carlo simulation results showed that with an appropriate illumination and fluorescence collection geometry, the distortion of <i>in vivo</i> fluorescence spectra of tissue caused by variations of optical properties at different wavelengths could be much reduced. Based on these results, a spectrofluorometry system was developed for the collection of <i>in vivo</i> laser-induced fluorescence spectra of tissue during endoscopy. In comparing the <i>in vivo</i> fluorescence spectral shape of bronchial tissue collected by this system with the intrinsic one obtained in vitro, we found no obvious distortion in the <i>in vivo</i> spectra. This was completely consistent with the analysis of Monte Carlo modeling. The <i>in vivo</i> measurement results demonstrated that significant differences in fluorescence intensity between normal and diseased bronchial tissue (dysplasia, carcinoma in situ) can be used to differentiate them from each other. Also, changes in fluorescence intensity are more robust for detecting abnormal tissues than the differences in spectral characteristics.

Immunohistochemical localization of cytochrome P450 2E1 in human pulmonary carcinoma and normal bronchial tissue: Kivisto KT, Linder A, Friedel G, Beaune P, Belloc C, Kroemer H et al. Dr Margarete Fischer-Bosch-Institut, Klinische Pharmakologie, Auerbachstrasse 112, D-70376 Stuttgart. Virchows Arch 1995; 426: 243–247

Immunohistochemical localization of cytochrome P450 2E1 in human pulmonary carcinoma and normal bronchial tissue: Kivisto KT, Linder A, Friedel G, Beaune P, Belloc C, Kroemer H et al. Dr Margarete Fischer-Bosch-Institut, Klinische Pharmakologie, Auerbachstrasse 112, D-70376 Stuttgart. Virchows Arch 1995; 426: 243–247

Immunohistochemical localization of cytochrome P450 3A in human pulmonary carcinomas and normal bronchial tissue: Kivisto KT, Fritz P, Linder A, Friedel G, Beaune P, Kroemer HK. Pathologisches Institut, Robert-Bosch-Krankenhaus, Auerbachstrasse 110, D-70376 Stuttgart. Histochem Cell Biol 1995;103:25–29

Immunohistochemical localization of cytochrome P450 3A in human pulmonary carcinomas and normal bronchial tissue: Kivisto KT, Fritz P, Linder A, Friedel G, Beaune P, Kroemer HK. Pathologisches Institut, Robert-Bosch-Krankenhaus, Auerbachstrasse 110, D-70376 Stuttgart. Histochem Cell Biol 1995;103:25–29

Immunohistochemical localization of cytochrome P450 2E1 in human pulmonary carcinoma and normal bronchial tissue.

Cytochrome P450 2E1 (CYP2E1) is a major xenobiotic-metabolizing enzyme but data concerning its extrahepatic expression are few. CYP2E1 can metabolically activate many procarcinogens and therefore its presence in the lung might play a role in bioactivation of procarcinogens, so we studied the expression and localization of CYP2E1 in primary pulmonary carcinomas and surrounding normal bronchial tissue from 28 patients. Seromucous glands showed expression of CYP2E1 in 19 and bronchial epithelium in 18 of the 28 samples of normal bronchial tissue. Thirteen of the corresponding cases of primary pulmonary carcinoma showed staining for CYP2E1. In 11 of these 13 cases, CYP2E1 was also present in normal bronchial tissue. There was no statistically significant difference in the expression of CYP2E1 between adenocarcinomas and squamous cell carcinomas. No association was observed between the expression of CYP2E1 in tumour tissue and normal bronchial tissue. However, there was a significant correlation between the expression of CYP2E1 in seromucous glands and bronchial epithelium (r = 0.61, P < 0.01) of normal tissue. We conclude that CYP2E1 can be present in both normal and neoplastic bronchial tissue.