Chemoresistance in
cancer therapy is an unfavorable prognostic factor in
non-small cell lung cancer (NSCLC). Elevation of intracellular
calcium level in multidrug resistant (MDR) sublines leads to sensitization of MDR sublines to cell death. We demonstrated that a fungal
protein from Ganoderma microsporum, GMI, elevates the intracellular
calcium level and reduces the growth of MDR subline via autophagy and apoptosis, regardless of
p-glycoprotein (P-gp) overexpression, in mice xenograft
tumors. In addition, we examined the roles of autophagy in the death of MDR A549
lung cancer sublines by GMI,
thapsigargin (TG) and
tunicamycin (TM) in vitro. Cytotoxicity of TG was inhibited by overexpressed P-gp. However, TM-induced death of MDR sublines was independent of P-gp level. Combinations of TG and TM with either
docetaxel or
vincristine showed no additional cytotoxic effects on MDR sublines. TG- and TM-mediated apoptosis of MDR sublines was demonstrated on
Annexin-V assay and Western blot and repressed by pan-
caspase inhibitor (
Z-VAD-FMK). Treatment of MDR sublines with TG and TM also augmented autophagy with accumulation of LC3-II
proteins, breakdown of p62 and formation of acidic vesicular organelles (AVOs). Inhibition of ATG5 by
shRNA silencing significantly reduced autophagy and cell death but not apoptosis following TG or TM treatment. GMI treatment inhibited the phosphorylation of Akt/S473 and
p70S6K/T389. Interestingly, the phosphorylation of ERK was not associated with GMI-induced autophagy. We conclude that autophagy plays a pro-death role in acquired MDR and upregulation of autophagy by GMI via Akt/mTOR inhibition provides a potential strategy for overcoming MDR in the treatment of
lung cancers.