Effect of GLT on the PhIP/DSS induced colon carcinogenesis and inflammation.

<p>Tumor incidence are summarized using percentage of animals with tumors and compared across groups using Fisher's exact test and the Bonferonni correction for multiple comparisons: ^a p<0.001 PhIP/DSS vs control, PhIP, DSS; ^b p<0.02 PhIP/DSS+500 GLT vs PhIP/DSS.</p><p>Tumor multiplicity are summarized using mean ± SD and compared across all group using Kruskal-Wallis one way analysis of variance on ranks and the Dunn's method for the multiple comparisons: ^a p<0.05 PhIP/DSS vs control PhIP, DSS; ^b p<0.05 PhIP/DSS+500 GLT vs PhIP/DSS.</p><p>Neoplastic index is summarized using median (min, max) and compared across groups using the Kruskal-Wallis test. Comparisons of each group to control performed using Mann-Whitney U tests with significance levels adjusted using the Bonferroni correction: ^a p<0.001 control vs PhIP; ^b p<0.001 control vs PhIP/DSS.</p><p>Data for colon length summarized using mean ± SD and compared across all group using ANOVA and Dunnett's post hoc test: ^ap<0.001 control vs DSS, control vs PhIP/DSS; ^bp<0.001 PhIP/DSS vs PhIP/DSS+100 GLT, PhIP/DSS vs PhIP/DSS+500 GLT.</p

GLT suppresses PhIP/DSS induced formation of colon tumors and inhibits focal hyperplasia and ACF formation.

<p>(A) Schematic of the animal treatment. The details of the treatment are described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. (B) Body weight of control animals (black circle), animals treated with PhIP (white square), DSS (black square), PhIP/DSS (white circle), PhIP/DSS+GLT 100 mg/kg of body weight (black triangle), and PhIP/DSS+GLT 500 mg/kg of body weight (white traingle) during the experiment. (C) H&E staining of representative samples from animal experiments described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#pone-0047873-g001" target="_blank">Figure 1A</a>. (D) Focal hyperplasia was evaluated by the histological analysis after H&E staining in colon tissue samples as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. Results are means ± SD (n = 6–9 mice/per group). (E) ACF formation was evaluated after methylene blue staining as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. Results are means ± SD (n = 10 foci/3 mice/per group), *p<0.05 by ANOVA.</p

Effect of GLT on the liver function and serum glucose.

<p>Values are Mean ± S.D. (n = 6), ALP, alkaline phosphatase; ALT, alanine transaminase; AST, aspartate transaminase. No significant difference from the control group.</p

GLT down-regulates expression of COX-2 in colon tissue.

<p>(A) Immunohistochemistry and (B) quantification of COX-2 positive cells were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. Box plots represent 5^th/10^th percentiles, horizontal bars represent median values, and whiskers indicate minimum to maximum values. Significant differences (*p<0.05) were observed among PhIP/DSS vs. control, PhIP/DSS vs PhIP/DSS+GLT 100, and PhIP/DSS vs. PhIP/DSS+GLT 100.</p

Effect of GLT on the lipid metabolism.

<p>Values are Mean ± S.D. (n = 6), HDL, high-density lipoprotein. No significant difference from the control group.</p

GLT activates XRE/AhR, PXR and rPXR. <i>in vitro</i>.

<p>Induction of (A) XRE/AhR, (B) PXR and (C) rPXR was evaluated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. Results are means ± SD (n = 4), *p<0.05 by ANOVA.</p

GLT down-regulates expression of Ki-67 and suppresses the amount of infiltrated macrophages.

<p>Immunohistochemistry and quantification of (A) Ki-67 and (B) F4/80 positive cells were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. Results are means ± SD (n = 4–5 mice/per group), *p<0.05 by ANOVA. Representative pictures from (A) Ki-67 and (B) F4/80 staining in the PhIP/DSS group and the PhIP/DSS group treated with GLT (500 mg/kg of body weight) 3 times/week for 120 days.</p

GLT prevents PhIP/DSS induced formation of colon tumors.

<p>(A) Schematic of the preventive experimental protocol. The details of the treatment are described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. (B) H&E staining of representative samples from animal experiments.</p

Effect of GLT on PhIP/DSS-induced expression of CYP1A2, CYP3A4 and CYP3A1 in colon tissue.

<p>Quantification of (A) CY1A2, (B) CYP3A4, and (C) CYP 3A1 positive cells as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047873#s4" target="_blank"><i>Materials and Methods</i></a>. Box plots represent 5^th/10^th percentiles, horizontal bars represent median values, and whiskers indicate minimum to maximum values. Significant differences (*p<0.05) were observed among (A) CYP1A2: PhIP/DSS vs. control, PhIP/DSS vs PhIP/DSS+GLT 100 (B) CYP3A4: PhIP/DSS vs. control, PhIP/DSS vs PhIP/DSS+GLT 100, and PhIP/DSS vs. PhIP/DSS+GLT 100, and (C) CYP3A1: PhIP/DSS vs. control.</p

Effect of GLT on the renal function.

<p>Values are Mean ± S.D. (n = 6), BUN, blood urea nitrogen. No significant difference from the control group.</p