ERK/MAPK-dependent beclin-1 expression is important for autophagic cell death induced by RAD001 and MK-2206.

<p>HGC-27 cells and AGS cells were treated for 24 h with RAD001 (10 nM) and/or MK-2206 (100 nM), phospho- and total levels of ERK/MAPK were tested by western blots (A). HGC-27 cells were pre-treated with MEK/ERK inhibitors PD98059 (PD, 1 µM) or U0126 (U, 1 µM) for 1 hour, followed by RAD001 (10 nM) and MK-2206 (100 nM) co-administration, 72 h later, cell viability was analyzed (B), the expression levels of beclin-1 and LC3B were also examined by western blots 48 h after administration. Scramble and beclin-1 siRNA transfected HGC-27 cells were treated with RAD001 (10 nM) and MK-2206 (100 nM) co-administration, cell viability (72 h after treatment) was measured by CCK-8 assay (D), and expression of LC3B, beclin-1, phospho- and total-MAPK were tested by western blots (E, after 48 h). p-MAPK (A and E), Beclin-1 (E) and LC3B (E) were quantified the same as above, and the number was normalized to the corresponding band of control. The data shown are the mean from three independent experiments. *<i>p</i><0.05.