Carcinogen‐Induced Animal Models of Head and Neck Squamous Cell Carcinoma

Daniel E. Bassi1, A.J.P. Klein‐Szanto1

1 Fox Chase Cancer Center, Philadelphia, Pennsylvania
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 14.2
DOI:  10.1002/0471141755.ph1402s37
Online Posting Date:  June, 2007
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This unit provides two models of oral carcinogenesis (hamster cheek pouch and rat tongue), as well as more simple procedures to induce squamous cell carcinomas (SCCs) in mouse skin. The most significant usage of these models has been their application in studying the molecular pathology of the genesis and multi‐step progression of tobacco‐associated SCC. Nevertheless, these protocols have been used frequently as in vivo bioassays to demonstrate the chemopreventive and, to a lesser extent, the chemotherapeutic effects of numerous compounds. Several examples, using four different carcinogenesis protocols, are presented with detailed steps on how to elicit squamous lesions and when and how to apply test compounds that could have either cancer chemopreventive or chemotherapeutic effects.

Keywords: carcinogenesis; animal models; head and neck cancer; chemoprevention; squamous cell carcinoma

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Table of Contents

  • Basic Protocol 1: The Hamster Cheek Pouch Carcinogenesis Model
  • Basic Protocol 2: Rat Tongue Cancer Model
  • Basic Protocol 3: Murine Surrogate Head and Neck Carcinogenesis Model: Two‐Stage Chemical Carcinogenesis of the Skin
  • Alternate Protocol 1: Complete Carcinogenesis Using Benzo(a)pyrene—“One‐Stage” Carcinogenesis
  • Alternate Protocol 2: Therapeutic Study Example: Transgenic Murine Surrogate Head and Neck Carcinogenesis Model—Two‐Stage Chemical Carcinogenesis of the Skin
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Basic Protocol 1: The Hamster Cheek Pouch Carcinogenesis Model

  • Male Syrian golden hamsters (6 weeks old; 80 to 180 g; Charles River or Harlan)
  • Hamster food: AIN‐93M diet (Research Diets)
  • 7,12‐dimethylbenz(a)anthracene (DMBA solution I; see recipe)
  • Mineral oil (Sigma)
  • Test compound
  • Disposable cages (Fisher Scientific)
  • Micropipettor with disposable plastic tips or paint brush (e.g., camel hair brush no. 4)
CAUTION: DMBA is a potent carcinogen and should be handled with care. Weigh and handle DMBA in a chemical fume hood (e.g., Multihazard Glove Boxes) dedicated for carcinogens, and use a spatula reserved for this specific purpose only. DMBA may be weighed into a glass scintillation vial, which is then capped immediately after use.

Basic Protocol 2: Rat Tongue Cancer Model

  • Male Fisher 344 rats (Charles River), 3 to 4 weeks old, 50 to 75 g body weight
  • Rat food
  • 4‐nitroquinoline 1‐oxide (4‐NQO solution; see recipe)
  • Test compound
  • Disposable cages and bottles (Fisher Scientific)
CAUTION: 4‐NQO is a potent carcinogen and should be handled with care. It should be weighed in a dedicated closed hood equipped with multihazard glove boxes and a spatula used for this specific purpose only. 4‐NQO may be weighed into a glass scintillation vial, which is then capped immediately after use. As with the other carcinogens described in this unit, use safety glasses, gloves, and good ventilation when handling 4‐NQO.

Basic Protocol 3: Murine Surrogate Head and Neck Carcinogenesis Model: Two‐Stage Chemical Carcinogenesis of the Skin

  • Female mice (6 to 8 weeks old; any of several lines and strains may be used, e.g., SENCAR and FVB × FVB (N); Taconic)
  • Mouse food
  • Bleach
  • DMBA solution II (see recipe)
  • Acetone (analytical grade, Fisher Scientific)
  • 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA solution; see recipe)
  • Test compound (optional)
  • Shaver and blades (e.g., Olsten Golden A5, equipped with an A5 clipper, size 40)
  • Disposable cages (Fisher Scientific)
  • 25‐ml vials
  • Digital camera
  • Caliper
CAUTION: DMBA is a potent carcinogen and should be handled with care. DMBA should be weighed in a dedicated closed hood equipped with multihazard glove boxes and with a spatula used for this specific purpose only. DMBA may be weighed into glass scintillation vials that are capped immediately after use. Concentrated (10×) solutions of DMBA can be prepared in a biosafety level 2 hood by dissolving DMBA in acetone. Mix by pipetting and/or with a rotating plate. Pipets that are used to handle carcinogens should not be used for other purposes and should be kept in the hood at all times. Pipet tips and excess DMBA solutions should be immediately decontaminated by the addition of three to five parts of bleach and disposed of as biohazardous material.

Alternate Protocol 1: Complete Carcinogenesis Using Benzo(a)pyrene—“One‐Stage” Carcinogenesis

  • Female FVB × FVB (N) mice (6 to 8 weeks old; Taconic)
  • Benzo(a)pyrene (B(a)P; see recipe)
  • Acetone (analytical grade; Fisher Scientific)
  • Bleach
  • Test compound
  • Disposable animal cages (Fisher Scientific)
  • Shaver with blades
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Literature Cited

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