Effect of Mouse Intestinal Bacteria on Incidence of Colorectal Tumors Induced by 1,2-dimethylhydrazine Injection in Gnotobiotic Transgenic Mice Harboring Human Prototype c-Ha-ras Genes

Abstract

We produced transgenic (Tg) gnotobiotic (GB) mice carrying human prototype c-Ha-ras genes and compared the incidence of colorectal tumors induced by 1,2-dimethylhydrazine (DMH) injection. At 7 to 11 weeks of age, germfree (GF) CB6F1-Tg Hras2 mice were inoculated with various mouse fecal suspensions or mixtures of bacteria isolated from mouse feces. Three weeks after bacterial inoculation, DMH was administered by subcutaneous injection at 20 mg per kg body weight for 20 weeks. Mice were euthanized 5 weeks after the last injection to investigate the number of colorectal tumors. The incidence of colorectal tumors was high in both Tg- and non-Tg-GF mice (100%). In Tg-specific pathogen-free (SPF) mice and Tg-GB-4 mice associated with basic mouse flora consisting of Escherichia coli, lactobacilli, Bacteroides and clostridia, the incidence of colorectal tumors was as high as that in GF mice. In Tg-SPF mice, the tumor score was higher than in Tg-GF mice (p<0.01), but no colorectal tumors were detected in non-Tg groups of SPF, and the tumor incidence was remarkably low in non-Tg-GB-4 mice. The tumor incidence and score in Tg- and non-Tg-GB mice varied depending on the bacterial combination in their intestine. These results indicate that the presence of human c-Ha-ras genes and intestinal bacteria substantially modify colorectal tumorigenesis induced by DMH.