Retinoic acid increases the expression of p53 and proapoptotic caspases and sensitizes keratinocytes to apoptosis: a possible explanation for tumor preventive action of retinoids.

Abstract

Retinoids influence growth and differentiation of keratinocytes (KCs) and are widely used for the management of skin diseases and for prevention of nonmelanoma skin cancer (NMSC) in predisposed patients. Here we investigated the effect of all-trans-retinoic acid (ATRA) on KC apoptosis. When KCs were cultured in confluent monolayers for several days, they acquired resistance against UVB-induced apoptosis. In contrast, when the cells were treated with 1 micromol/L ATRA for 6 days and subsequently irradiated with different doses of UVB, they underwent massive apoptosis as assessed by morphology, expression of activated caspase-3, and DNA fragmentation. The same effect was observed when doxorubicin was used instead of UVB. Analysis by real-time PCR and Western blot revealed that ATRA treatment strongly increased the mRNA and protein expression of p53 and caspase-3, -6, -7, and -9, which are key regulators of apoptosis. UVB irradiation of ATRA-treated cells but not of control cells led to the accumulation of p53 protein and of its target gene Noxa. Inhibition of p53 and caspases with alpha-pifithrin and z-Val-Ala-Asp-fluoromethyl ketone, respectively, blocked UVB- and doxorubicin-induced apoptosis in ATRA-treated KCs. Analogous to the observed ATRA effects in monolayer cultures, in vitro-generated organotypic skin cultures reacted with up-regulation of p53 and proapoptotic caspases and displayed increased sensitivity to UVB-induced apoptosis. The ability of retinoic acid to regulate the expression of proapoptotic genes and to sensitize KCs to apoptosis may play a role in their prevention of NMSC in transplant patients and patients with DNA-repair deficiencies.

• Authors: Paul Mrass, Michael Rendl, Michael Mildner, Florian Gruber, Barbara Lengauer, Claudia Ballaun, Leopold Eckhart, Erwin Tschachler
• Journal: Cancer research (Cancer Res) Vol. 64 Issue 18 Pg. 6542-8 (Sep 15 2004) ISSN: 0008-5472 [Print] United States
• PMID: 15374966 (Publication Type: Journal Article)

Chemical References

• Antibiotics, Antineoplastic
• Caspase Inhibitors
• Isoenzymes
• Tumor Suppressor Protein p53
• Tretinoin
• Doxorubicin
• Caspases

Topics

• Antibiotics, Antineoplastic (pharmacology)
• Apoptosis (drug effects, physiology, radiation effects)
• Caspase Inhibitors
• Caspases (biosynthesis)
• Cells, Cultured
• Down-Regulation (drug effects)
• Doxorubicin (pharmacology)
• Drug Synergism
• Humans
• Isoenzymes
• Keratinocytes (cytology, drug effects, enzymology, radiation effects)
• Skin Neoplasms (metabolism, prevention & control)
• Tretinoin (pharmacology)
• Tumor Suppressor Protein p53 (antagonists & inhibitors, biosynthesis)
• Ultraviolet Rays
• Up-Regulation