Activations of Both Extrinsic and Intrinsic Pathways in HCT 116 Human Colorectal Cancer Cells Contribute to Apoptosis through p53-Mediated ATM/Fas Signaling by Emilia sonchifolia Extract, a Folklore Medicinal Plant
Figure 6
ESE-affected cytotoxicity and apoptosis is mediated through alterations of p53 downstream signals in p53 siRNA-transfected HCT 116 cells. (a) The p53 siRNA or control siRNA-transfected HCT 116 cells were treated with ESE (50 μg/mL) for 12 h, and total protein was prepared and subjected to Western blotting analysis. The membranes were incubated with anti-p53, anti-Fas, anti-PUMA, anticaspase-8 and, anti-caspase-3 antibodies. The blot was also probed with anti-Actin antibody to confirm equal loading of samples. Each band was quantified using ImageJ software. The p53 siRNA or control siRNA-transfected HCT 116 cells were treated with ESE (50 μg/mL) for 24 h, cell viability was determined by MTT assay (b) and apoptotic cells were assessed by TUNEL assayand flow cytometric analysis (c). The values presented are the mean ± S.D. (n = 3) from three independent experiments. ***P < 0.001 shows a significant different when compared to control sample.
