Early detachment of colon carcinoma cells during CD95(APO-1/Fas)- mediated apoptosis. I. De-adhesion from hyaluronate by shedding of CD44

doi:10.1083/jcb.134.4.1089

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Early detachment of colon carcinoma cells during CD95(APO-1/Fas)- mediated apoptosis. I. De-adhesion from hyaluronate by shedding of CD44

AR Gunthert, J Strater, U von Reyher, C Henne, S Joos, K Koretz, G Moldenhauer, PH Krammer and P Moller
Institute of Pathology, University of Heidelberg, Germany.

Ligation of CD95 (APO-1/Fas) cell surface receptors induces death in apoptosis-sensitive cells. Induction of apoptosis in adherent gamma interferon-stimulated HT-29 and COLO 205 colon carcinoma cells by cross- linking CD95 with anti-APO-1 monoclonal antibody resulted in detachment of the cells from hyaluronate starting about 1 h after antibody exposure. Loss of adhesion was paralleled by a substantial reduction of the multifunctional cell surface adhesion molecule CD44. As evidenced by cycloheximide treatment, this effect was not caused by impaired protein synthesis. Depletion of surface CD44 was also not due to membrane blebbing, since cytochalasin B failed to inhibit ascension from hyaluronate. Instead, ELISA and time kinetics showed increasing amounts of soluble CD44 in the supernatant of CD95-triggered cells. SDS- PAGE revealed that soluble CD44 had an apparent molecular mass of about 20 kD less than CD44 immunoprecipitated from intact cells. Thus, CD95- triggering induced shedding of CD44. Shedding is a novel mechanism operative in early steps of CD95-mediated apoptosis. Shedding surface molecules like CD44 might contribute to the active disintegration of dying epithelial cells in vivo.
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