Multi-stage chemical carcinogenesis in mouse skin: fundamentals and applications

Nat Protoc. 2009;4(9):1350-62. doi: 10.1038/nprot.2009.120. Epub 2009 Aug 27.


For more than 60 years, the chemical induction of tumors in mouse skin has been used to study mechanisms of epithelial carcinogenesis and evaluate modifying factors. In the traditional two-stage skin carcinogenesis model, the initiation phase is accomplished by the application of a sub-carcinogenic dose of a carcinogen. Subsequently, tumor development is elicited by repeated treatment with a tumor-promoting agent. The initiation protocol can be completed within 1-3 h depending on the number of mice used; whereas the promotion phase requires twice weekly treatments (1-2 h) and once weekly tumor palpation (1-2 h) for the duration of the study. Using the protocol described here, a highly reproducible papilloma burden is expected within 10-20 weeks with progression of a portion of the tumors to squamous cell carcinomas within 20-50 weeks. In contrast to complete skin carcinogenesis, the two-stage model allows for greater yield of premalignant lesions, as well as separation of the initiation and promotion phases.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Carcinogens / toxicity
  • Carcinoma, Squamous Cell / chemically induced*
  • Disease Models, Animal*
  • Mice
  • Papilloma / chemically induced*
  • Skin Neoplasms / chemically induced*


  • Carcinogens