Effect of Ganoderma applanatum polysaccharides on MAPK/ERK pathway affecting autophagy in breast cancer MCF-7 cells

Fungal polysaccharides have become hotspots in the field of health foods due to their antitumor activity in recent years. In this experiment, antitumor effect of the medicinal fungus Ganoderma applanatum polysaccharide (GAP) was investigated in human breast cancer MCF-7 cells, as well as the molecular mechanism of its effect on autophagy. Results showed that GAP contains three polysaccharides with molecular weights of 6.36 × 10⁵ Da, 4.25 × 10⁵ Da, and 2.53 × 10⁵ Da and which composed of rhamnose, glucose, arabinose, fucose in the molar ratio of 1:22:16.1:3.2. GAP inhibited the proliferation and migration of MCF-7 cells in a time-dose-dependent manner, the maximum inhibition rate reached 50.2% at 500 μg/mL in 48 h. Flow cytometry analysis showed that GAP could induce apoptosis, treatment of cells with GAP could result in up-regulation of gene and protein levels of autophagy-associated markers LC3 and Beclin-1; addition of autocrine late inhibitor CQ significantly raised the protein expression level of LC3II. The mitogen-activated protein kinases (MAPK) signaling pathway was not only related to the apoptotic pathway but also to the autophagy pathway; Western blot analysis showed that MAPK signaling pathway is involved in GAP-induced autophagy in MCF-7 cells. Detection of the relevant signaling pathway protein showed that the expression of p-ERK1/2 protein was down-regulated, however the expression of p-p38 and p-JNK protein was up-regulated. These results indicate that GAP could induce early autophagy in MCF-7 cells via the MAPK/ERK pathway. In conclusion, GAP showed strong antitumor activity by inducing apoptosis and autophagy through MAPK signaling pathway in MCF-7 cells, suggesting the molecular mechanism of fungal polysaccharide on its antitumor activity.