Anti-tumor effect of polysaccharide from Hirsutella sinensis on human non-small cell lung cancer and nude mice through intrinsic mitochondrial pathway.

Our previous works had proved the structural properties of Hirsutella sinensis polysaccharide-III(HSP-III). Herein, its anti-tumor effect on lung cancer correlated with mitochondrial apoptosis pathway was investigated. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that HSP-III induces the apoptosis of H1299 cells; however the proliferation viability of normal lung epithelial cells is not affected. HSP-III treatment collapses the H1299 cell mitochondrial membrane potential, and western blot analysis of cytochrome C, Bax, caspase-3 and caspase-9 further indicates that apoptotic effects induced by HSP-III is through the mitochondrial pathway. Furthermore, we found the apoptotic effects of HSP-III are triggered by Reactive oxygen species (ROS) generation. Blue native Polyacrylamide Gel-Electrophoresis (PAGE) showed the expressions of mitochondrial respiratory chain complexes I-V were also decreased. Taken together, anti-tumor effect of HSP-III is through intrinsic mitochondrial apoptosis mechanism pathway and involving ROS increasing. Finally, in vivo nude mice experiment, HSP-III attenuated the growth of tumor compared with control. In contrast, N-acetyl-l-cysteine (NAC) could restore the cell apoptosis effects induced by HSP-III. These findings suggest that HSP-III induce apoptosis of H1299 cells and attenuated growth of nude mice tumor in vivo through the intrinsic mitochondrial pathway and stimulating ROS. HSP-III could be a composition for lung cancer treatment.