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Cyclooxygenase-independent induction of p21WAF-1/cip1, apoptosis and differentiation by L-745,337, a selective PGH synthase-2 inhibitor, and salicylate in HT-29 cells

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Abstract

In order to dissect out cyclooxygenase-dependent from cyclooxygenase-independent mechanisms in the antiproliferative effects of selective prostaglandin H synthase (PGHS)-2 inhibitors, we compared the effects of L-745,337 (a highly selective PGHS-2 inhibitor) with sodium salicylate (a weak PGHS inhibitor) on prostanoid production, induction of the cyclin-dependent kinase inhibitor p21WAF-1/cip1, mutant p53 (m273-p53) levels, apoptosis and differentiation in human colon adenocarcinoma HT-29 cells. L-745,337 dose-dependently suppressed the cyclooxygenase activity of HT-29 cells (IC50: 0.24 μM). Four-day treatment with L-745,337 caused a concentration-dependent inhibition of cell growth (IC50: 0.9 mM) associated with the induction of p21WAF-1/cip1 and an increase in the proportion of apoptotic nuclei (EC50: 0.1 and 0.34 mM, respectively) while reducing the levels of m273-p53 (IC50: 0.2 mM). Sodium salicylate, at the concentration of 10 mM that did not affect prostanoid formation, caused a 60% reduction of cell growth associated with a 3-fold induction of p21WAF-1/cip1 and a 60% increase in the proportion of apoptotic nuclei. Ultrastructural analysis showed that L-745,337 (0.5 mM) and sodium salicylate (10 mM) caused the induction of a differentiated phenotype. We conclude that high concentrations of L-745,337 and sodium salicylate inhibit colon cancer cell growth by a mechanism unrelated to cyclooxygenase inhibition that may involve p53-independent induction of the tumor suppressor p21WAF-1/cip1.

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Authors and Affiliations

  1. Departments of Medicine and Aging, Division of Pharmacology, Italy

    G. Santini, M. G. Sciulli, A. Ricciardulli & P. Patrignani

  2. Institut National de la Santè et de la Recherche Médicale, Unité 348, Hopital Lariboisière, 75475, Paris, France

    R. Marinacci & A. Pace

  3. IFR Biologie de la Circulation-Lariboisiére, Institut National de la Santè et de la Recherche Médicale, Unité 348, Hopital Lariboisière, 75475, Paris, France

    J. Maclouf

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  1. G. Santini
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  3. R. Marinacci
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  4. O. Fusco
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  5. L. Spoletini
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Santini, G., Sciulli, M.G., Marinacci, R. et al. Cyclooxygenase-independent induction of p21WAF-1/cip1, apoptosis and differentiation by L-745,337, a selective PGH synthase-2 inhibitor, and salicylate in HT-29 cells. Apoptosis 4, 151–162 (1999). https://doi.org/10.1023/A:1009631204581

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