Methotrexate induces apoptotic cell death in human keratinocytes.
The mechanism by which low doses of methotrexate act in psoriasis to restore a clinically normal skin is poorly understood. Apoptosis is a programmed cell death activated when cell removal is needed. The purpose of the present work was to examine using an organotypical model of keratinocyte culture, the possibility that low doses of methotrexate can induce apoptosis of keratinocytes. Epidermal explants were cultivated on dead deepidermized dermis under air-exposed conditions. After 10 days, methotrexate (10(-7) M) was added. After a further 5 days, one part of each culture was fixed and submitted to routine histology, DNA nick end labelling (TUNEL) to detect DNA fragmentation (a molecular marker of apoptotic cell death) and immunohistochemical detection of p53 (a protein involved in apoptosis induced by DNA-damaging agents). The other part of each culture was processed for electron microscopy. A significant proportion of keratinocytes (1%) were damaged and exhibited the morphological features of apoptotic cell death. Immunohistochemical overexpression of p53 was detected in the basal layer of the cultures treated with methotrexate. Low doses of methotrexate induce apoptosis. This mode of action could explain the reduction in epidermal hyperplasia during treatment of psoriasis with methotrexate.
Department of Dermatology, Hôpital Erasme, Université Libre de Bruxelles, Belgium.
M
M
Heenen
M
M
Laporte
J C
JC
Noel
C
C
de Graef
D017209:Apoptosis / Q000187:drug effects*; D002452:Cell Count / Q000187:drug effects; D002467:Cell Nucleus / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D003879:Dermatologic Agents / Q000494:pharmacology*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D008727:Methotrexate / Q000494:pharmacology*; D011565:Psoriasis / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D012867:Skin / Q000166:cytology / Q000187:drug effects / Q000473:pathology; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000187:drug effects
D003879:Dermatologic Agents; D016159:Tumor Suppressor Protein p53; D008727:Methotrexate