Fig. 6

Effects of sulindac sulfide on miR-27a and ZBTB10 in colon cancer cells. a Sulindac sulfide decreases miR-27a promoter activity in SW480 and RKO cells. Cells were transfected with luciferase construct containing −638 bp promoter sequence of miR-27a gene (pMir27a-Luc). After 6 h, cells were treated with DMSO, 75, 100 and 125 μM sulindac sulfide for 18 h and luciferase activity was determined as described under Materials and Methods. b Sulindac sulfide decreases miR-27a transcript level in SW480 and RKO cells and this effect is reversed by antioxidant NAC. Cells were treated with DMSO, 100 μM sulindac sulfide alone or in combination with 10 mM NAC for 24 h and cellular level of the miR-27a transcript was determined by TaqMan PCR analysis as described under Materials and Methods. c Sulindac sulfide increases gene expression of ZBTB10 in SW480 and RKO cells. Cells were treated with DMSO, 25, 50 and 75 μM sulindac sulfide for 24 h and ZBTB10 mRNA levels were determined by real-time PCR analysis as described under Materials and Methods. All experiments were done in triplicate and results are expressed as fold of control (mean ± S.D.). Asterisk (*) indicates statistical difference between control (DMSO) and treatment, and double-asterisk (**) indicates statistical difference between single treatment and combination treatment (P < 0.05). d Effects of sulindac sulfide on the ROS-miR-27a-ZBTB10-Sp axis. Sulindac sulfide induces ROS; downregulates miR-27a; upregulates ZBTB10; downregulates Sp proteins and Sp-dependent survival/proliferative, inflammatory and angiogenic protein products; and induces growth inhibition and apoptosis