Figure 6

The effects of digitoflavone on colon carcinogenesis. (A) Schematic overview of digitoflavone administration. Colons were removed at week 13 after the mice were administered digitoflavone or a vehicle control between week 2 and 13. (B) The tumor numbers, sizes and colon length were determined macroscopically. The bars represent the median of each group. Each symbol represents the tumor numbers and colon length of each animal or the average size of the tumors of each animal. Significantly different: *p<0.05, **p<0.01. (C) Macroscopic evaluation of the tumors. Colons were removed on week 13 from mice, treated with digitoflavone or with vehicle. Representative results from 6 independent animals are shown here. Original magnification, ×6. (D) Colons were processed for hematoxylin and eosin staining and representative results from 6 independent animals are shown here. Original magnification, × 40. (E) Immunoblotting analysis with anti–Nrf2, TR, γ-GCSc, γ-GCSm and HO-1 antibodies was performed on cell lysates from colon tissues as described in Methods. Representative results from 3 independent experiments are shown here (left panel), the intensity of bands from replicate immunoblots was quantified and plotted (right panel, bar graphs). Significantly different (versus control group): *p<0.05, **p<0.01. (F-G) Quantitative RT-PCR analysis for GR, TR, HO-1, γ-GCSc, γ-GCSm, NQO-1, UGT1A1, UGT1A10, TNF-a, IL-1βand IL-6 was performed on total RNAs extracted from the colons. All genes’ mRNA levels were normalized to the levels of GAPDH mRNA. Significantly different (versus control group): *p<0.05, **p<0.01 and NS, not significant.