NSC 95397 is a potent inhibitor of Cdc25 dual specificity phosphatase(Ki of 32, 96, and 40 nM for Cdc25A, Cdc25B, and Cdc25C, respectively).

CDC25B:96nM(ki), CDC25C:40 nM(ki), CDC25A:32nM(ki)

NSC 95397 reduced cell viability and anchorage-independent growth of all the three colon cancer cell lines through inhibited proliferation and induced apoptosis via regulating cell-cycle-related proteins, including p21, cyclin-dependent kinases, and caspases[1].

To measure the cell proliferation activity of NSC 95397 against colon cancer cells, SW480 (1 × 10^4), SW620 (2 × 10^4), and DLD-1 (1 × 10^4) cells were seeded into 96-well plates.?After overnight culture, NSC 95397 cells were added at the indicated concentrations.?After 24 h of incubation, cell proliferation was determined in vitro using a BrdU cell proliferation assay kit.?O.D. 450 values were analyzed by using a Multiskan PC.The apoptosis of colon cancer cells was determined by a PE Annexin V Apoptosis Detection Kit with 7-AAD.?Briefly, cells were treated with the indicated concentration of NSC 95397 for 24 h. After 24 h, Cells were harvested and stained with PE Annexin V/7-AAD for 15 min. The stained cells were analyzed using FACSCanto II low cytometer and FCS Express software[1]