Immunocytochemical expression of the pi class glutathione S-transferase (GST) was investigated in preneoplastic and neoplastic lingual lesions in a 4-nitro-quinoline 1-oxide (4NQO)-induced rat genetic model [Wistar/Furth rats (WF) and Dark-Agouti rats (DA)] and in human surgical material [fibrous polyp, mild to moderate dysplasia, severe dysplasia, carcinoma in situ (CIS), squamous cell carcinoma (SCC)]. Two polyclonal antibodies raised against rat (GST-P) and human (GST-pi) antigens were used. In the rat model, DA and WF rats showed contrasting susceptibility to 4NQO, DA rats having a much higher tumour incidence and a significantly shorter survival time than WF rats. While the established lingual SCC in DA and WF rats all expressed GST-P, the number of GST-P+ foci in the preneoplastic lingual epithelium was significantly higher in DA (14.5 +/- 6.5) than in WF rats (5.5 +/- 2.6; P < 0.0001). In contrast, GST-pi epithelial staining in human specimens was more variable and the results overlapped in different groups. More frequent nuclear and/or basal cell staining was detected in severe dysplasia, CIS and SCC than in benign and mild to moderate dysplastic lesions. Although the pi class GST may be a useful marker for rat lingual carcinogenesis, its value in clinical applications is unclear. GST-pi staining patterns and their distribution may be helpful in identifying high-risk lingual lesions in humans.