Light-induced fluorescence spectroscopy was conducted on human oral malignant and normal tissues. Under 330-nm excitation wavelength, significant differences in fluorescence intensity were observed around 380- and 460-nm emission. Furthermore, 7,12-dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis in hamster buccal pouch was investigated to elucidate whether similar alterations of fluorescence spectroscopy occurred during the development of squamous cell carcinoma. Similar to the spectral profiles of human oral malignant and normal tissues, the most intense fluorescence peaks in the pouches occurred at 380 nm and 460 nm emission under 330 nm excitation wavelength. At 380 nm emission, the fluorescence intensity of normal pouch mucosa was stronger than those of DBMA-treated abnormal tissues at different stages of carcinogenesis. However, at 460 nm emission, the fluorescence intensity of DMBA-treated tissues was not only stronger than that of normal pouch mucosa but also shifted to 470 nm. These results suggest that under 330 nm excitation wavelength fluorescence spectroscopy may be useful for the detection of oral malignant lesions.