Two-Dimensional Electrophoresis of Proteins in Tumours of the Lung

Abstract

The proteins of solid lung tumours (15 adenocarcinomas and 10 squamous cell carcinomas) were examined by high resolution two-dimensional electrophoresis (2-DE) and compared with the proteins of adjacent lung tissue which demonstrated no histological evidence of malignant transformation, and with the proteins of other malignant tumours and normal tissues. To investigate tumour cell-specific protein synthesis, we isolated malignant and normal cells enzymatically with collagenase, elastase, and DNase. Tissue and tumour cells were enriched in an additional Step on a Percoll gradient. The 2-DE gel patterns derived from entire tissue and enriched tissue cell preparations were compared.No specific differences were found between the 2-DE protein patterns from adenocarcinomas and squamous cell carcinomas of the lung, but three proteins identified on the 2-DE gels appeared to be tumour-associated. Spot A is present in non-neoplastic and neoplastic epithelial tissues. Spot B is pronounced in 2-DE gels of sarcomas, but is also present in preparations of other malignant tissues. Spot C is present in all malignant cell preparations. These three spots were also demonstrated in 2-DE protein patterns from tissue cultures of mälignaiit cell lines. Spot B and spot C were also present in some normal tissues. Zweidimensionale Elektrophorese von Proteinen in LungentumorenZusammenfassung: Solide Lungentumoren (15 Adenocarcinome und 10 squamöse Zellcarcinome wurden mit der hochauflösenden zweidimensionalen Elektrophorese (2-DE) untersucht. Diese Tumoren wurden mit dem angrenzenden Lungengewebe, das keine histologischen Hinweise auf maligne Transformationen zeigte und mit anderen malignen Tumoren und normalen Geweben verglichen. Um die tumorspezifische Proteinsynthese zu studieren, isolierten wir maligne und normale Zellen aus soliden Tumoren und Geweben enzymatisch mit Kollagenase, Elastase und DNase. In einem zusätzlichen Schritt wurden Gewebe-und Tumorzellen auf einem PereoU-Gradienten angereichert. 2-DE-Gele vom gesamten Gewebe und angereicherten Gewebezellpräparationen wurden verglichen.EndJer et al.: Two-dimensional electrophoresis of proteins in lung tumour Die Adenocarcinome und squamösen Zellcarcinome der Lunge wiesen in der 2-DE keine spezifischen Unterschiede auf. Drei tumor-assoziierte Proteine konnten auf den Gelen identifiziert werden. Fleck A befindet sich in nicht-neoplastischen und neoplastischen epithelialen Geweben. Fleck B kommt insbesondere in 2-DE-Gelen von Sarkomen, aber auch in anderen malignen Geweben vor. Fleck C zeigt sich in 2-DE-Gelen aller maligner Zellpräparationen. Diese drei Proteinflecke ließen sich auch in 2-DE-Gelen von Zellkulturen maligner Zellinien nachweisen. Fleck B und C fanden sich auch in einigen normalen Geweben.

Two-dimensional electrophoresis of synovial tissue, synovial fluid and serum in patients with rheumatoid arthritis and related diseases

“…Likewise, 2D electrophoresis has proved to be a powerful tool for protein studies o f almost every h u m a n body fluid and tissue, e.g. seminal fluid [26], h u m a n brain tissue [27], brain t u m o u r tissue [28], lung t u m o u r tissue [29], uterine tissue [30], tissue taken from patients with undifferentiated leukaemia [31], and urine [32,33]. The present study was aimed at the detection of disease-associated changes in the 2D protein patterns of serum, synovial fluid and synovial tissue extracts obtained from patients suffering from RA or non-RA.…”

Two-dimensional cryostat section electrophoresis: a novel method and its application to the diseased synovial membrane

“…Two-dimensional electrophoresis followed up by silver staining or immunoblotting of human body fluids and tissue is now a well-established technique used to characterize the protein pattern of normal and diseased persons either for diagnostic or scientific purposes (Anderson & Anderson, 1977, 1978Edwards et aI., 1981;P6hling & Neuhoff, 1981;Lonberg-Holm et al, 1982;Manabe et al, 1982Manabe et al, , 1985Tracy et al, 1982;Jackson & Thompson, 1984;Kossmann, 1984;Steisslinger, 1985;Endler & Young, 1986;Guest et al, 1986;Hanash & Baier, 1986;Narayan et al, 1986;Wiederkehr et al, 1986;Peristeris et al, 1987;Marshall et al, 1988;Choukaife eta]., Menzel & Unteregger, 1989;Kovalyov et al, 1990). In rheumatology there are so far only rare examples of its application.…”

Two‐dimensional polyacrylamide gel electrophoresis of human lung cancer: Qualitative aspects of tissue preparation in relation to histopathology

“…Technical improvements are required regarding the preparation of solid tumor tissues in order to generate 2-DE patterns of higher quality. Few studies on solid lung cancers using 2-DE have been made so far [4,6] and the preparation techniques used vary. In general, extremely diverse methods have been proposed for the preparation of solid tumor tissues.…”

“…The main disadvantage of these methods is the difficulty in removing serum Abbreviations: DDT, dithiothreitol; 2-DE, two-dimensional polyacrylamide gel electrophoresis; IEF, isoelectric focusing; Mr, molecular weight; pl, isoelectric point proteins and necrotic material. This problem might be solved by enzymatic extraction and separation of whole cells before adding 2-DE lysis buffer [6]; however, it is not easy to fully control nonspecific digestion of cell membrane proteins. On the other hand, the use of tissue culture techniques might change the expression of critical polypeptides.…”