Effects of LM-1685, a non-coxib COX-2 inhibitor, on the viability and induction of ER stress-related molecules in NTUB1 and T24 cells.

<p>(A) Cells were treated with LM-1685 (0–160 µM) for 24 h. Cell viability was analyzed by MTT assay. Data are presented as means ± SD of three independents experiments. (B) Cells were treated with LM-1685 (80 and 160 µM) or celecoxib (100 µM) for 24 h. The cell lysates were harvested at each time point and analyzed by Western blotting with specific antibodies to detect ER stress-related molecules IRE-1α, CHOP, calnexin, and GRP78, and caspase-4. CF is the abbreviation of cleaved form. (C). Cells were transfected with GRP78 siRNA (10 nM) or scramble siRNA (10 nM) (as a control), and then treated with 160 µM LM-1685. The combinative effect of LM-1685 and GRP78 knockdown on apoptosis was determined by flow cytometry (FACS) with annexin V-FITC and PI labeling. The quantitative analysis of total apoptosis (early and late) population was presented. Data are presented as means ± SD of three independents experiments. * p<0.05 as compared with scramble siRNA+LM-1685.