Figure 2

MA potentiates TNFα-induced pancreatic cancer cell apoptosis and suppresses TNFα-induced invasion. (A) MA inhibits TNFα-induced cell viability by Live/Dead assay. Panc-28 cells were pretreated with or without 25 microM MA for 12 h, and then incubated with 0.1 nM TNFα for 24 h. The live (green) or dead (red) cells were stained with Live and dead assay kit as mentioned in Materials and Methods. Magnification, ×10. (B) MA potentiates cell apoptosis induced by TNFα using annexin V staining (top) and TUNEL staining (bottom). Panc-28 cells were pretreated with or without 25 microM MA for 6 (or 12) h, and incubated with or without 0.1 nM TNFα for another 12 (or 24) h, then stained with annexin V-FITC (or TUNEL) and analyzed by flow cytometry as mentioned in Materials and Methods. (C) MA potentiates the cleavage of PARP and caspase-3. Panc-28 cells were pretreated with or without 25 microM MA for 6 h, and then incubated with or without 0.1 nM TNFα for the indicated time. Cell lysates were harvested and analyzed by Western blot with the indicated antibodies. (D) MA suppresses pancreatic cancer cell invasion induced by TNFα. Panc-28 cells were seeded onto the top chambers of the transwell. After treated with 10 microM MA for 12 h, the number of cells invaded the transwell was counted as described in Materials and Methods. Columns, mean; bars, SD. Magnification, ×10.