A responsive, sensitive, and reproducible dermal tumor promotion assay for the comparative evaluation of cigarette smoke condensates

The mouse dermal initiation/promotion bioassay has been used for several decades to study cigarette smoke condensates (CSCs). However, these studies have used highly variable methodologies that differ in the manner of CSC collection, duration of treatment, mouse strain, number of mice and endpoints measured. In this report, a protocol that uses female SENCAR mice and standardizes many of the procedures is presented. A reference cigarette (University of Kentucky 1R4F), readily available to researchers, was used. This report presents the combined data from four independent studies. Female, SENCAR mice (40/group) were treated with a single dose (75 μg) of dimethylbenz[a]anthracene (DMBA) as an initiator, followed 1 week later by treatment (three times/week) with 10, 20 or 40 mg “tar”/application of 1R4F CSC for 29 weeks. There were no treatment-related effects on body weights. Histological diagnosis of all masses at study termination indicated a dose-dependent increase in the number of tumor-bearing mice and total tumor number. These studies support the conclusion that the 1R4F cigarette is suitable for use as a reference standard and the protocol presented is an appropriate and standardized model suitable for the comparative evaluation of CSC.