SM-164 promotes APO2L/TRAIL-induced apoptosis and inhibits AKT activation in human HCC cell lines.

<p>(a–b). SMMC-7721 cell line was treated with SM-164 (SM) at 0.1 µM alone, APO2L/TRAIL (A/TR) at 100 ng/ml alone or both for 16 h, cells were stained with Annexin V/PI staining and examined by flow cytometry assay. (a). Representative dot plot of apoptosis assay. (b). data of Annexin V-positive cells show means ± S.D. of from three experiments. **, p<0.01. (c). SMMC-7721 cell line was treated as indicated, whole cell lysates were analyzed with western blotting and specific antibodies as indicated. β-Actin was used as a loading control. (d). SMMC-7721 cell line was pretreated with pan-caspases inhibitor, zVAD (50 µM) for 1 h and followed by treatment with SM at 0.1 µM alone, A/TR at 100 ng/ml alone or both for 24 h, cell death induction was examined with trypan blue exclusion assay. (e). BEL-7402 cell line treated with SM at 0.1 µM alone, A/TR at 10 ng/ml or both for 16 h was examined with Annexin V/PI staining and flow cytometry assay. (f). HepG2 cell line treated with SM at 0.1 µM alone, A/TR at 1000 ng/ml or both for 16 h was examined with Annexin V/PI staining and flow cytometry assay. (g-h). BEL-7402 and HepG2 cell lines were treated as indicated for 12 h. Whole cell lysates were analyzed with western blotting and specific antibodies. (i). Cell lines were treated as indicated for 12 h. Whole cell lysates were analyzed with western blotting and specific antibodies. β-Actin was used as a loading control.