Figure 4
Effects of EpCAM mAb on the induction of apoptosis in colon cancer cells analyzed by DAPI and TUNEL staining. Apoptotic cell death was determined by DAPI staining and TUNEL assay. Treatment with anti-EpCAM mAb (10 µM) and RAW 264.7 cells for 24 h caused apoptosis; characterized by marked chromatin condensation, small membrane-bound bodies (apoptotic bodies), cytoplasmic condensation, and cellular shrinkage. (A) The TUNEL-positive (green) cells are a apoptotic cells (200× magnification). Apoptotic cells (DAPI-stained TUNEL-positive cells) were estimated by direct counting of fragmented nuclei after DAPI and TUNEL staining. SW620 colon cancer cells (2 × 106/ml) either untreated or treated with doses of anti-EpCAM mAb (10 µM) and RAW264.7 cells. An equal amount of total protein (40 µg/lane) was subjected to 12% SDS-PAGE. (B) Expression of Bax, caspase-3, caspase-8, Bcl-2, TNF-α, and β-actin were detected by western blotting using specific antibodies. Here, β-actin protein was used as an internal control. Each band is representative of 3 independent experiments. Values represent mean (SD) of 3 experiments, each performed in triplicate. *P < 0.05 indicates a statistically significant difference from the control group.
