EP1.03-27 The Anti-Migration and Anti-Invation Mechanisms of Capilliposide C from Lysimachia Capillipes on Lung Cancer

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Approximately 50% of lung cancer patients had distant metastases when initial diagnosed. Lysimachia capillipes is one of traditional medicine in China. It is proved to be safety in clinical use. Several studies have showed the antitumor effects of Capilliposide from Lysimachia capillipes (LC) in vitro. Our preliminary data showed LC could inhibit the migration and invasion of lung cancer. This study aims to explore the detailed mechanisms of LC as well as its extracts on lung cancer. Four non-small cell lines were selected. The invasive response of lung cancer cells was determined by Wound Healing assay. Proteins and the phosphorylation of proteins were evaluated by iTRAQ-based proteomics. The phosphorylation chip was used to evaluate the phosphorylation effect. PC-9 xenografts were used to evaluate the antitumor of LC-C in vivo. Immunohistochemistry (IHC) was used to evaluate the antitumor mechanisms in vivo. The migration capa city of lung cancer cells was significantly reduced after treatment of LC-C. Proteomics showed there were 364 differentially expressed proteins and 456 differentially expressed phosphorylated proteins in both LC-C treated PC-9 and H1975 cells. Differentially expressed proteins were enriched in EGF receptor signaling pathway, Wnt signaling pathway, Cadherin signaling pathway, Notch signaling pathway, TGF-beta signaling pathway, and p38 MAPK pathway by bioinformatic analysis. Phosphorylation chip showed LC reduced the phosphorylation of AKT, WNK1 and PRAS40, but not EGFR. Western blot showed the phosphorylation of mTOR, AKT and PRAS40 were significantly inhibited after LC-C treatment; besides, the inhibitory effects of AKT and mTOR were dose-dependent. While the total proteins of AKT and mTOR were not changed. Western blot showed the phosphorylation of Smad2 and Smad3 were significantly inhibited after LC-C treatment. Western blot showed E-Cadherin was up-regulated and N-Cadherin was down-regulated after LC-C treatment; while other EMT related proteins including ZO-1, Snail and Vimentin were not changed. Nor did cell adhesion related protein Claudin-1. PC-9 xenograft model showed the tumor growth inhibitory rates were 114.4% in 7 days after LC-C administration. IHC showed the phosphorylation of AKT was down-regulation after LC-C administration, Ki-67 was also down-regulation, while cleaved caspase-3 was not changed. The study showed LC-C inhibit the growth and the capacity of invasion and migration of lung cancer cells. The detailed mechanisms might crosstalk with several critical pathway such as AKT pathway, TGF-β pathway and EMT.