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Molecular & Cellular Proteomics 1:269-279, 2002.
© 2002 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Proteomic Strategies to Reveal Tumor Heterogeneity among Urothelial Papillomas ^*

Julio E. Celis^,,¶, Pamela Celis, Hildur Palsdottir^,||, Morten Østergaard, Pavel Gromov^,, Hanne Primdahl^**, Torben F. Ørntoft^**, Hans Wolf, Ariana Celis^, and Irina Gromova^,

Institute of Cancer Biology and Danish Centre for Human Genome Research, The Danish Cancer Society, DK-2100 Copenhagen
Department of Medical Biochemistry, The University of Aarhus, Ole Worms Allé, Building 170, DK-8000 Aarhus
^** Departments of Clinical Biochemistry
Urology, Skejby Hospital, DK-8200 Aarhus N, Denmark

Proteomics and immunohistochemistry were used to reveal tumor heterogeneity among urothelial papillomas (UPs) with the long term goal of predicting their biological potential in terms of outcome. First, we identified proteins that were deregulated in invasive fresh lesions as compared with normal urothelium, and thereafter we immunostained UPs with a panel of antibodies against some of the markers. Twenty-two major proteins showing variations of 2-fold or more in at least one-third of the invasive lesions were selected. Specific antibodies against several of the proteins were obtained, but only a few reacted positively in immunostaining. A panel consisting of antibodies against keratinocytes (CKs) 5, 13, 18, and 20 and markers of squamous metaplasia (CKs 7, 8, and 14) was used to probe normal urothelium and 30 UPs collected during a period of five years. Four UPs showed a normal phenotype, whereas the rest could be grouped in five major types that shared aberrant staining with the CK20 antibody. Type 1 heterogeneity (n = 4) showed preferred staining of the umbrella cells with the CK8 antibody. Type 2 (n = 11) was typified by the staining of the basal and intermediate layers with the CK20 antibody. Type 3 (n = 7) was characterized by the predominant staining of the basal cell layer with the CK5 antibody. Type 4 (n = 1) showed areas of CK7 negative cells, whereas type 5 (n = 3) showed loss of staining of the basal cells with the CK20. 29% of the patients experienced recurrences, but none progressed to invasive disease. Patients harboring phenotypic alterations in the basal cell compartment (types 3 and 5) showed the highest number of recurrences (4/7 and 2/3, respectively), and all type 3 lesions progressed to a higher degree of dedifferentiation. Even though a long term prospective study involving a larger sample size is required to assess the biological potential of these lesions, we believe that this approach will prove instrumental for revealing early phenotypic changes in different types of cancer.

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