Article thumbnail

Cell surface antigens in renal tumour cells: detection by immunoluminescence and enzymatic analysis

By F Laube, B Göhring, H Sann and I Willhardt

Abstract

Two renal cell carcinoma cell lines (49RC 43STR and 75RC 2STR) were characterized by detection of the cell surface proteins: CD44(var), intercellular adhesion molecule-1 (ICAM-1), urokinase-type plasminogen activator (uPA) and its receptor and aminopeptidase N (APN). To detect their localization the immunoluminescent technique was used. In addition, the enzyme activity of uPA and APN was investigated in cell suspensions as well as in monolayers. The latter procedure was more advantageous since the additional use of HPLC permits a single registration of the fluorescent hydrolysis-product AMC (7-amino-4-methylcoumarin) without interference by cellular autofluorescence or non-reacted fluorescent substrate. Unlike 75RC 2STR, the cell line 49RC 43STR expressed high levels of uPA and APN. Contrary to that the cell line 75RC 2STR expressed high levels of ICAM-1 and CD44(v6), whereas 49RC 43STR showed a low level of ICAM-1 and no distinct light signal with anti-CD44(v6). The uPA activity was measured directly as well as indirectly (via plasmin) with the substrate Z-Gly-Gly-Arg-AMC. Both activator and plasmin activity were inhibited by D-Val-Phe-Lys-CMK and phenylmethylsulfonyl fluoride. The anti-catalytic antibody to uPA and that to uPA receptor were found to be inhibiting the uPA activity in a concentration-dependent manner. APN activity was assayed using alanine-p-nitroanilide. Peptidase activity was effectively inhibited by 1,10-phenanthroline and partly inhibited by ethylenediamine-tetraacetic acid. © 2001 Cancer Research Campaignhttp://www.bjcancer.co

Topics: Regular Article
Publisher: Nature Publishing Group
OAI identifier: oai:pubmedcentral.nih.gov:2375078
Provided by: PubMed Central

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.

Suggested articles

Citations

  1. (1991). Academic Press: San Diego Evans CW
  2. (1996). Correlation of very late activation integrin and CD44 expression with extrarenal invasion and metastasis of renal cell carcinomas. Human Pathol 27: 1355–1360
  3. (1997). Molecular regulation of intercellular adhesion molecule 1 (ICAM-1) expression in renal cell carcinoma.
  4. (2001). Pathol 151: 1655–1661 927 BJOC 01-2004 924-929 10/9/01 2:00 pm Page 928Immunophenotyping of renal tumour cells 929
  5. (1997). The urokinase-type plasminogen activator system in cancer metastasis: A Review.