Gastrin in human bronchogenic carcinomas: constant expression but variable processing of progastrin.

Using a library of radioimmunoassays against essential sequences of human progastrin and procholecystokinin, we have examined the occurrence of gastrin, cholecystokinin, and their precursors in bronchogenic adenocarcinomas, large-cell, small-cell, and squamous-cell carcinomas (n = 17). Progastrin and some of its bioactive (i.e., alpha-carboxyamidated) products were present in all tumors, irrespective of histological classification. The concentration of progastrin varied from 0.2 to 21.9 pmol/g tissue; glycine-extended intermediates constituted less than 0.1 to 0.5 pmol/g; and bioactive, carboxyamidated gastrin ranged from less than 0.1 to 6.1 pmol/g. Chromatography showed that the bioactive gastrins were exclusively gastrin-17 peptides, half of which were tyrosine O-sulfated. Neither procholecystokinin nor its processing products were found in the tumor extracts. Six samples of nonneoplastic human lung tissue contained traces of progastrin (range, less than 0.1-0.8 pmol/g), but neither bioactive gastrins nor any cholecystokinin. The results show that the gastrin gene is expressed in all classes of bronchogenic carcinomas. Due to incomplete posttranslational processing measurement of progastrin may be necessary to detect such expression.

Soluble proteins of human bronchogenic carcinomas.

Solid-phase affinity chromatography has been used to search for tumor-associated proteins of bronchogenic carcinomas. All of the apparently normal proteins were removed from extracts of squamous cell carcinoma, large cell carcinoma, and adenocarcinoma of the lung. The remaining soluble proteins were partially characterized as to heat stability, approximate molecular size, electrophoretic mobility, and biologic function. These tumor-associated proteins were not tumor-specific by current definition. Neither lung cancer patients' lymphocytes nor serum proteins were specifically reactive with these tumor-associated proteins.

Usefulness of tissue polypeptide antigen in staging, monitoring, and prognosis of lung cancer.

A study of tissue polypeptide antigen (TPA) serum levels was conducted in patients with various cell types of bronchial carcinoma. A total of 548 assays (226 pretreatment) were evaluated. In all, the sensitivity of the test at diagnosis was 61 percent. It varied considerably in relation to the extent and site of disease. Individual pretreatment values of TPA correlated significantly with the stage of disease. Similarly, posttreatment TPA values related to disease evolution. In small cell carcinoma, eight of the 29 meaningful changes in follow-up levels of TPA preceded clinical recognition of disease status variations. Raised pretreatment values of TPA were significantly associated with a shortened survival. These results indicate that TPA may contribute to staging, monitoring, and prognosis of lung cancer.

Microsequencing evidence for the maturation of human proopiomelanocortin into an 18 amino acid beta-melanocyte stimulating hormone [h beta MSH(5-22)] in nonpituitary tissue.

Sixty pmoles of a material with molecular size, immunological, and RP-HPLC characteristics identical to that of h beta MSH(5-22) were purified from a bronchial carcinoid tumor responsible for the ectopic ACTH syndrome. The first 16 cycles of microsequencing revealed the following sequence: Asp-Glu-Gly-Pro-Tyr-Arg-Met-Glu-X-Phe-Arg-Trp-Gly-X-Pro- Pro-, identical to the first 16 amino acids of h beta MSH(5-22). Since this material was recognized by an antibody which requires the free COOH-terminal Asp22 residue, it can be assumed that it is indeed h beta MSH(5-22). We also show that neither the 5 N acetic acid nor the 1 N HCl extraction procedure artefactually generated h beta MSH-like material in normal or tumoral human pituitaries and in nonpituitary tumors. We conclude that h beta MSH(5-22) is a normal maturation product of proopiomelanocortin in the human nonpituitary tissues which express its gene, including the hypothalamus and ACTH-secreting tumors.

Serum ferritin in patients with cancer: determination with antibodies to HeLa cell and spleen ferritin.

Some malignant tissues and cell lines contain acidic isoferritins and it has been suggested that the assay of such isoferritins in serum may be of value in the diagnosis of malignancy. This paper describes a radioimmunoassay for acidic ferrtin purified from HeLa cells. Examiniation of purified heart, kidney, liver and spleen ferritin showed that the assay was highly specific for acidic isoferritins. Ferritin concentrations have been measured with antibodies to HeLa cell and spleen ferritin in extracts of normal and tumour tissue. Although the tumours contained more HeLa type ferritin than the corresponding normal tissue the HeLa/spleen type ferritin ratio was low. HeLa-type ferritin concentrations have been compared with values obtained with anti-spleen ferritin in over 1000 sera from normal subjects and patients with cancer and leukaemia. HeLa-type ferritin as not detected ( less than 2 micrograms/l) in most normal sera. Concentrations of up to 53 micrograms/l were found in sera from patients with malignant disease but the HeLa/spleen type ferritin ratio was always very low. There appears to be little application for antibodies to HeLa cell or heart ferritin in the diagnosis or monitoring of cancer.

Immunohistochemical studies of keratin in human bronchus and lung tumors.

Lung tumors of various types, fixed in 4% formaldehyde-1% glutaraldehyde, were stained for keratin proteins. The results were compared with previous ultrastructural evidence of intermediate filament bundles (IFBs), presumed to be keratin. Electron and light microscopic methods were largely complimentary and the results were in agreement in 79% of cases. Light microscopy was superior for demonstrating keratinizing foci containing numerous well-developed IFBs, whereas electron microscopy was superior when keratin filaments were sparsely distributed in cells throughout a tumor. Fetal and adult bronchial specimens were also studied. Normal adult bronchus, fixed in aldehydes, was unreactive but keratin was observed in similarly fixed bronchi that showed epidermoid metaplasia and/or intraepithelial carcinoma. Keratin was demonstrated in normal adult bronchi fixed in ethanol. Keratin was not observed in fetal lung until the 14th week of gestation, when it appeared in basal cells and a few columnar cells of the larger bronchi. Thereafter, keratin progressively appeared in the more distal branches. As specimens from gestations of less than 14 weeks were fixed in aldehydes, the apparent lack of immunoreactivity may have been artifactual. Nevertheless, keratin was demonstrable in aldehyde-fixed fetal bronchi at 16 and 23 weeks' gestation.

Bronchoalveolar T-lymphocytes and subsets in pulmonary tuberculosis and bronchogenic carcinoma.

Analysis of T-lymphocytes and their subsets in the blood and bronchoalveolar lavage fluids from 12 patients with active pulmonary tuberculosis, 12 patients with bronchogenic carcinoma and 11 healthy volunteers was aimed at identifying their immunologic functions and interrelationships. In patients with active pulmonary tuberculosis, there was a significant increased in the percentage of T-cells bearing IL-2 receptor both in the blood and bronchoalveolar lavage fluids, whereas patients with bronchogenic carcinoma exhibited an increase in the suppressor T-cells and T-cells bearing IL-2 receptor in the blood only. The presence of T-cells bearing IL-2 receptor is generally accepted to be the hallmark of recently active specific antigen activation of the helper T-lymphocytes together with monokine IL-1 stimulation. Suppressor T-cells, on the other hand, play a role in the immunopathogenesis of lung cancers and lung metastasis.

Analysis of lung air from patients with bronchogenic carcinoma and controls using gas chromatography-mass spectrometry.

Volatile metabolites present in expired lung air were collected by odor sampling techniques and analyzed by gas chromatography-mass spectrometry. The study population included controls matched for age and smoking history with patients newly diagnosed with lung carcinoma. Significantly greater concentrations of o-toluidine were found in the lung air of patients with lung carcinoma than either age-matched or younger controls. Aniline was present in half of the patient population but absent in age-matched controls.

Analysis of the cores of ferruginous (asbestos) bodies from the general population. I. Patients with and without lung cancer.

Ferruginous (asbestos) bodies may be found in the lungs of almost everyone in the population, but little information is available as to whether such bodies are nucleated on asbestos or on some other fibrous dust. In this study morphologically "typical" ferruginous bodies were isolated from the lungs of 23 autopsy and surgical patients, none of whom had primary asbestos exposure. Eleven patients had carcinoma of the lung. To determine the nature of the core, 328 bodies were examined by electron diffraction. Of these, 264 (80%) showed the diffraction patterns of amphibole asbestos, whereas six showed the pattern of chrysotile asbestos. No amorphous cores or crystalline nonasbestos cores were identified. Fifty-eight (18%) bodies could not be diffracted because of the thickness of the iron-protein coat. No differences were seen between patients with and without lung cancer. We conclude that typical ferruginous bodies have asbestos cores, which are usually amphibole type. The findings suggest widespread exposure to asbestos dust; occupational histories appeared to indicate the source of exposure in some but not all patients.

Association of asbestos and bronchogenic carcinoma in a population with low asbestos exposure.

Quantitative counts of ferruginous bodies were performed on digests of lungs from 100 control and 30 lung cancer patients. It was found that the lung cancer group had significantly higher levels, although only 1 patient was known to be occupationally exposed to asbestos. It is suggested that even extremely low levels of asbestos exposure may have a carcinogenic effect.

Quantitative DNA determinations by image analysis. I. Application to human pulmonary cytology.

A description is made of the application of image analysis to quantitative deoxyribonucleic acid (DNA) determinations in a variety of cells. Optical density of the video signal from Feulgen-stained material was integrated across the area of the nucleus and values obtained related to values derived from known diploid cells. Analysis was performed on three classes of material: (1) biological material with known haploid, diploid and tetraploid nuclear DNA (standards), (2) metaplasias of the bronchial epithelium and (3) bronchogenic carcinomas. Results obtained with image analysis corresponded with expected data for the biologic standards and with microspectrophotometric data obtained for metaplasia and epidermoid carcinoma of the bronchial system. Application of this and other quantitative methods to cancer screening and biology is discussed.

Asbestos bodies in the lung: Southampton (UK) and Wellington (New Zealand).

Two series of surgically removed and unselected necropsy lungs were examined for asbestos bodies. In the series from Southampton (UK) 30-mu sections were used and in the Wellington (New Zealand) series a modified Gold's digestion method was used for all cases, together with 30-mu sections for some. A further series of Southampton lungs was assessed by Gold's method six years after the initial study. The 30-mu section technique found asbestos bodies in 13% of Southampton men and none in women: the corresponding Wellington figures were 8% for men and nil for women. In Wellington the more sensitive digestion method found asbestos bodies in 78% of men and 63% of women. In Southampton the investigation by Gold's method found asbestos bodies in 83% of men and 74% of women. No statistically significant association was found between the number of asbestos bodies and the various types of lung carcinoma. The prevalence of asbestos in lungs removed between 1956 and 1965 did not differ significantly from its prevalence in those removed in the late 1970's. There was no significant increase in asbestos after the age of 20.

Clinical value of a multiple biomarker assay in patients with bronchogenic carcinoma.

In 98 newly diagnosed patients with histologically proven bronchogenic carcinoma seen at Cuneo Hospital of Chest Diseases from July 1983 to December 1984, multiple biomarker assays were performed. Fiftynine cases had more than one carcinoembryonic antigen (CEA) and/or tissue polypeptide antigen (TPA) assay during the course of the disease, at 3- to 12-week intervals. A total of 209 CEA (91 pretreatment), 170 TPA (80 pretreatment), 62 human chorionic gonadotropin (HCG)-beta subunits and 60 lactate dehydrogenase (LDH) was assayed. In addition, serum samples were taken from 141 blood donors and their TPA values were used as a control. The percentages of elevated values were, respectively, 37%, 52%, 18%, and 25%. In 85% of the patients at least one biomarker was found to be higher than normal. Neither significant differences between mean biomarker levels in tumors of various histologic types nor positive intermarker correlations were found. The number of patients with elevated CEA, TPA, and LDH serum levels and their mean values increased significantly according to the disease extent. Among evaluated markers TPA showed the highest accordance to tumor burden. The raising of two markers was never associated with Stage I-II disease, except in one patient. Both CEA and TPA concentrations changed significantly during the course of the illness in relation to the clinical status assessment. Abnormal pretreatment levels of CEA, LDH, and particularly, TPA were independently and significantly associated with a poor outcome. Patients with abnormal levels of TPA and LDH and, to a lesser degree, TPA and beta-HCG had shorter survival as compared with patients with high TPA values, irrespective of the LDH and beta-HCG levels, although not significantly so.

Flow-cytometric assessment of DNA index and percent S phase cells in bronchogenic epidermoid carcinoma.

The DNA index and percent S phase cells were assessed in 45 bronchogenic epidermoid carcinomas from paraffin-embedded retrieved material by flow cytometry. Previously, a comparison between flow cytometric findings from fresh and from paraffin-embedded material was made. The DNA index and the percent S phase cells from fresh and paraffin-embedded material were strongly correlated. Only 3 of the 45 cases were diploid. The DNA index was significantly lower in grade 1 than in grade 2-3 tumors. No correlations were found between DNA index and presence of lymph node metastases, but the percent S phase cells was significantly higher in lymph-node-positive tumors.

[Significance of alpha-1-antitrypsin and alpha-2-pregnancy-associated glycoprotein in the serum of patients with bronchial carcinoma]. / Die Bedeutung des Alpha-1-Antitrypsins und des Alpha-2-schwangerschaftsgebundenen Glykoproteins im Serum von Kranken mit Bronchialkarzinom.

The authors investigated alpha-1-antitrypsin and pregnancy associated alpha-2-glycoprotein at diagnosis and follow-up of patients with bronchogenic carcinoma. Both proteins were determined by single radial immunodiffusion according to Mancini in 60 patients with bronchogenic carcinoma, in 31 patients with nontumorous respiratory diseases, and in 10 patients with tumour metastases in the lungs. The statistical significance of differences was evaluated using Student's t-test. None of the determined proteins was found to be a specific and sensitive marker of bronchogenic carcinoma. The concentration of alpha-1-antitrypsin is increasing with the growth of the tumour, and the values of pregnancy associated alpha-2-glycoproteins are decreasing at the same time. Alpha-1-antitrypsin can be used in follow-up after tumour resection, where recurrent increase of its concentration may indicate a relapse of the tumour.

Quantitative DNA determination by image analysis. II. Application to human and canine pulmonary cytology.

Comparative DNA measurements in human and canine preneoplastic and neoplastic tracheobronchial cells were made with the application of computerized image analysis. Canine studies demonstrated that the sequence of cellular events that precede epidermoid lung cancer simulates precisely the progression observed in humans. DNA studies concomitantly confirmed that there is a stepwise increase in DNA content with advancing nuclear atypia in metaplastic respiratory cells in both species. All carcinomas, regardless of histologic type, were significantly hyperploid to aneuploid (4c to 6c). Small-cell carcinoma exhibited a narrow modal distribution in the 4c region. The uniformity of the cytologic and quantitative DNA changes among these disparate species tends to confirm that humans and canines share biologic characteristics in bronchogenic carcinogenesis. The quantitative DNA measurements provide an objective cellular marker and may be used clinically for diagnostic purposes.

Tumor markers in bronchogenic carcinoma. An evaluation of carcinoembryonic antigen, tissue polypeptide antigen, placental alkaline phosphatase and pseudouridine.

From 62 patients with bronchogenic carcinoma, carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), placental alkaline phosphatase (PLAP) in serum and pseudouridine, a modified nucleoside, were analysed in urine. About 60 per cent of the patients had squamous cell carcinoma, and 20 per cent had small cell carcinoma. The patients were allocated into 3 different clinical stages based upon tumor burden, and the markers were analysed before treatment and thereafter. TPA and PLAP had limited value as biologic markers. For both CEA and pseudouridine the frequency of elevated values increased parallel to clinical stage. Elevated levels of these 2 markers were also correlated to shorter survival.

Immunohistochemical localization of carcinoembryonic antigen (CEA), CEA-S, and nonspecific cross-reacting antigen (NCA) in carcinomas of lung.

Antisera to carcinoembryonic antigen (CEA), to a physicochemical subset of CEA, namely CEA-S, and to nonspecific cross-reacting antigen (NCA) were used for the immunohistochemical localization of these antigens in human bronchogenic carcinomas using a triple layer immunoperoxidase technique. The study is based on an analysis of tumors from 130 patients. CEA, CEA-S, and NCA were all identified in the membrane and/or cytoplasm of neoplastic cells, and a good correlation between the antigens was observed in a majority of tumors. The presence or absence of these tumor-associated glycoproteins appeared to be correlated with the histologic type of the tumors, especially in small cell anaplastic carcinoma and adenocarcinoma, and the degree of histologic differentiation of adenocarcinomas correlated positively with these tumor-associated antigens. Data from this group of patients suggest that analysis of tissue CEA at the time of biopsy or surgical resection may facilitate a more objective interpretation of serial plasma CEA assays.

Subretinal fluid examination of LDH, PGI, and CEA in a case of metastatic bronchogenic carcinoma of the choroid.

A 58-year-old white male with a history of bronchogenic carcinoma presented with a total retinal detachment overlying a choroidal metastasis. His main tumor burden had been extirpated by pneumonectomy followed by radiation therapy four months prior to admission. With the development of a painful, glaucomatous eye, unresponsive to conventional therapy, enucleation was performed. Histologic examination of the enucleated globe revealed a metastatic tumor to the choroid, consistent with primary bronchogenic carcinoma. Aqueous humor and plasma examination revealed elevated ratios (Aqueous humor:Plasma) of lactate dehydrogenase (LDH) and phosphoglucose isomerase (PGI). Furthermore, subretinal fluid examination demonstrated concentrations of LDH and PGI higher than aqueous humor. While the level of carcinoembryonic antigen (CEA) in the plasma was normal (less than 2.5 ng/ml) following pneumonectomy, it was 121 ng/ml in the subretinal fluid. This would suggest that a choroidal lesion alone, in the absence of a clinically detectable primary tumor, is insufficient to elevate the plasma CEA.