We have carried out a study of large malignant skin tumors (squamous cell carcinomas) and other lesions in hairless mice (groups of 38-45) intermittently exposed to ultraviolet light over a period of 15 weeks, beginning when the mice were about 10 weeks old. The several groups were given a standard diet with 0%, 0.3%, 5%, and 10% added L-ascorbic acid (vitamin C) throughout the study. No lesions developed in unirradiated control groups. The lesions were counted every 14 days for 4 months, beginning 4 weeks before the end ofthe period ofirradiation. The observed incidence of lesions of several sizes during successive time periods was analyzed by the statistical method recommended by a committee of the International Agency for Research on Cancer. A pronounced effect of vitamin C in decreasing the incidence and delaying the onset of the malignant lesions was observed with high statistical significance.Interest in vitamin C and other dietary factors in relation to the incidence of and mortality from cancer in human beings has been stimulated by the studies of the value of vitamin C for patients with advanced cancer in Vale of Leven Hospital (Loch Lomondside, Scotland) from 1971 on (1-9). In 1973 it was suggested to one of us (L.P.) that we carry on studies of vitamin C in relation to cancer in animals (8). We began these studies in 1976, and we recently published a report on four studies of vitamin C and vitamin E in relation to skin cancer in hairless mice exposed to ultraviolet light in a standard way (10). In the present paper we report on another study on irradiated mice.About 10 years ago, Homer Black and his collaborators reported that cholesterol-5d,6d-epoxide (cholestan-5d,6d-epoxy-3-ol), a carcinogenic sterol, is formed in skin on ultraviolet irradiation, both in humans (11) and in mice (12, 13). They also reported that dietary anti-oxidants [a mixture ofL-ascorbic acid, butylated hydroxytoluene, D,L-a-tocopheryl acetate (vitamin E), and glutathione] suppressed tumor formation induced in hairless mice by exposure to ultraviolet light (12). Black has given us much information and advice about his techniques.
MATERIALS AND METHODSThe hairless mouse is a mutant that loses its hair about 3 weeks after birth, providing easy visibility of skin lesions. It does not have the thymus defect of the nude mutant. Our mice, all females, were of the SK h-hr strain bred at Temple University Health Sciences Center (Philadelphia). They were obtained in March and April of 1980 at age 6-9 weeks. They were maintained for at least 2 weeks on Purina certified rodent chow no.5002 to stabilize the colony before the special diets were begun.They were then distributed five mice to a cage so that the combined weight and mean age per cage were standardized (individual weights, 17.6-32.9 g; ages, 8-11 weeks). Irradiation was begun 10 days later and was continued 5 days per week for about 15 weeks to a total exposure of 135 J/cm2. The radiation was from GE-UA3 mercury arc lamps as described by Black except that an automatic timi...