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Ploidy, expression of erbB1, erbB2, P53 and amplification of erbB1, erbB2 and erbB3 in non-small cell lung cancer. N. Reinmuth, B. Brandt, W-P. Kunze, K. Junker, M. Thomas, R. Achatzy, H.H. Scheld, M. Semik. #ERS Journals Ltd 2000. ABSTRACT: The aim of this study was to assess the prognostic value of deoxyribonucleic acid analysis, expression of erbB1, erbB2 and P53, and amplification levels of erbB1, erbB2 and erbB3 in non-small cell lung cancer (NSCLC).Consecutive patients with NSCLC who underwent treatment with curative intention (118) were included. In 108 cases, the cell cycle was analysed using flow cytometry and double-staining with propidium iodide and anticytokeratin. In another 108 cases, expression of erbB1, erbB2 and P53 was assessed immunhistochemically. Amplification of the erbB family was determined in the tumours of 53 patients using doubledifferential polymerase chain reaction.Of the tumours, 81% were aneuploid and 14% showed positive staining for erbB1, 18% for erbB2 and 41% for P53. There were normal mean gene copy numbers in 86% for erbB1, 94% for erbB2 and in 96% for erbB3. No significant correlations were noted between erbB1, erbB2 and P53 expression, ploidy status and tumour stage. In a Cox regression model, only tumour stage was shown to be prognostically significant.It seems that ploidy and expression status of erbB1, erbB2 and P53 are not prognostic parameters in non-small cell lung cancer. Amplification of the erbB family does not seem to be a frequent event in non-small cell lung cancer. Eur Respir J 2000; 16: 991±996.

Morphometry is well-established in tumour pathology. To evaluate is potential usefulness for description of developmental processes, histological slides from paraffin-embedded specimens of 67 human fetal lungs were Feulgen-stained, and morphometric characteristics of nuclei of epithelial pulmonary cells were analysed with an automated image analysis system. The measured cytometric features comprised of integrated optical density (IOD), S-phase-related IOD fraction, IOD entropy and nuclear area. Histometric features of the specimens were based upon the minimum spanning tree (MST) and included distances between neighboring epithelial cells, between epithelial cells and neighboring lymphocytes, and assessment of MST entropy. Notably, certain parameters revealed a non-uniform level during prenatal development S-phase-related IOD fraction increased from 5% to 8% between 14 and 16 weeks of gestation, then declined to 6% until birth. The IOD entropy steadily increased during development, whereas the extent of nuclear area remained constant. In accordance with an increase of the S-phase-related fraction the MST entropy displayed a singular peak between 14 and 16 weeks of gestation, which is probably associated with development of glandular structures in the lung. Correlation of expression of binding sites for markers, presumably involved in functional aspects of development, with such alterations, is shown for binding capacities of biotinylated fucoidan and the S-phase-related fraction. This may be helpful to infer immuno- or ligando histochemically defined tissue sites with potential physiological significance in morphometrically distinguished periods of development.

Protein-carbohydrate interactions are supposed to play key roles in the mechanisms of cell adhesion, biosignalling and intracellular routing, warranting the analysis of the developmental course of expression of epitopes of this system. Thus, a panel of carrier-immobilized carbohydrate ligands was used as probes, namely lactose,N-acetylgalactosamine,N-acetylglucosamine, mannose, fucose and maltose. Additionally, an antibody to an endogenous β-galactoside-binding lectin (anti-galectin-1), the biotinylated lectin and two further human lectins, namely the macrophage migration inhibitory factor-binding sarcolectin and serum amyloid P component (SAP) that displays selectivity for sulphated sugars and mannose-6-phosphate, were included. They enabled us to assess the extent of the presence of respective binding sites in fixed sections from human lungs (pulmonary epithelial cells), livers (hepatocytes) and hearts (myocard cells) of 10-50 weeks gestation. Invariably, specific binding was detected in the three organ types, at least in certain stages. In most of the cases, the intensity of staining exhibited developmental regulation. The apparent patterns reveal similarities between the different cell types, as seen with immobilizedN-acetylglucosamine as well as with labelled galectin-1 and sarcolectin. However, drastic differences among such patterns with nearly opposite developmental courses do also occur, as detected for carrier-attached mannose and maltose residues. These results point to a potential importance for the detected glycohistochemical features in human development and substantiate the possibility of differential regulation of the presence of binding sites for distinct sugars within a certain organ and between the individual cell types of the monitored organs.

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