Figure 1. BITC inhibits the growth of in vivo xenograft by inhibiting the PI3K/AKT/FOXO pathway.

Athymic nude mice were kept on antioxidant free diet for one week before starting the experiment. Exponentially growing one million BxPC-3 cells were injected into both right and left flanks of each animal in PBS:Matrigel suspension. When tumors reached 70mm³ mice were randomly divided into two groups with ten mice in each group. Treated group received 12μmol BITC by oral gavage everyday while control group received vehicle alone. Tumors were measured three times a week. Effect of BITC on tumor volume (A), tumor weight (B) and mice body weight (C) was evaluated. The mechanism of tumor growth inhibition by BITC was determined in the tumor lysates from control and BITC-treated mice (D). Tumors were homogenized, lysed and 40μg protein was resolved on SDS-PAGE and probed for p-PI3K (Tyr-458), PI3K, p-AKT (Ser-473), AKT, p-mTOR (Ser-2448), mTOR, p-FOXO1 (Ser-256), FOXO1, p-FOXO3a (Ser-253), FOXO3a, Bim, PCNA, cleaved Caspase-3 and cleaved PARP. The blots were stripped and reprobed for actin to ensure equal protein loading. Lower (left) panel shows the quantitation of tumor western bolts. Values in (A and B) are means ± SEM of 20 samples and values in (C and D) are means ± SD of 10 samples.* P<0.05 statistically significant when compared with control.