Figure 8
From: Characterization of the biological activity of a potent small molecule Hec1 inhibitor TAI-1
Efficient P53 knockdown in cancer cells increases cellular sensitivity to TAI-1. (A) A549 and HCT116 cells which carry wild-type P53 were transfected with control siRNA (siControl) or P53 siRNA (siP53) for 24 hours and treated with TAI-1 (starting dose 100 μM, 3x serial dilution), incubated for 48 hours and analyzed for viability with MTS. Cellular sensitivity is expressed in GI50 (nM) and RNA from transfected cells were analyzed for P53 RNA level by quantitative real time PCR. SiP53 reduced GI50s of compound in cells. (B) Selected cell lines which carry wild type P53 (A549, HCT116, ZR-75-1, U2OS) or mutated P53 (HeLa, as control) were transfected with siP53, treated with TAI-1 and analyzed for viability with MTS. Cellular sensitivity is expressed as% growth inhibition and cell lysates from transfected cells were collected and P53 protein levels determined by western blotting.