FIGURE 6.

Role of BITC-induced ROS on the Myc-miR-ZBTB cascade. A and B, L3.6pL, MiaPaCa2, and Panc1 cells were pre-treated with 5 mm GSH for 3 h and then treated with 10 μm BITC alone or in combination with GSH, and after 6 h, total RNA was extracted. miR-17, miR-20a, and miR-27a (A) and ZTBT4, ZBTB10, and ZBTB34 (B) expressions were determined by real time-PCR. C, Panc1 cells were treated with 10 μm BITC for the indicated times, and ZBTB4, ZBTB10, and ZBTB34 proteins were analyzed by Western blotting. Cells were transfected with siCtl or two oligonucleotides targeting the sic-Myc#1 and sic-Myc#2 Western blottings of knockdown efficiency (D), and the effects of 10 μm BITC on proliferation of pancreatic cancer cells were determined in the presence or absence of c-Myc knockdown (E and F). Results shown in A, B, E, and F are expressed as mean ± S.E. for at least three replicate experiments, and significant (p < 0.05) changes by BITC (*) or reversal by GSH (#) are indicated in A and B. E and F, both BITC and c-Myc knockdown (sic-Myc) decrease cell proliferation (*).