Artesunate Represses The Growth And Metastasis Of Pancreatic Cancer Cells

Objective: Pancreatic cancer is a deadly solid malignancy.Thus,efficient drugs are urgently required because traditional chemotherapy does not provide satisfactory clinical outcomes.Artesunate(ART,artesunate)has been found to exhibit remarkable anti-tumor activity,thereby making it a novel candidate for cancer therapy.However,the anti-tumor mechanisms of ART have not been fully explored.This study is aimed at investigating the significance and mechanisms of the anti-tumor activity of ART against pancreatic cancer cells in vitro and in vivo.Methods: 1.Proliferation of pancreatic cancer cells treatment with ART was investigated by MTT assay.2.Pancreatic cancer xenograft model was established by subcutaneous injection of pancreatic cancer PANC-1 cell line into nude mice.3.The expression of CD34 in pancreatic cancer cells was detected by immunohistochemistry.4.Using RNA-Seq analysis to detect the expression of the targeted gene.5.Gene Ontology(GO)analysis was used to detect the main functions of differentially expressed genes(DEGs).Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis was used to analyze the important pathways of DEGs.6.Application of network pharmacology-based websites such as PyMOL and SystemsDock to predict the combination of ART and PP2 Ac.7.Flow cytometry was used to detect apoptosis.8.Caspase-8 and Caspase-9 activity assays were used to analyze the pathway of ART-induced apoptosis in pancreatic cancer cells.9.Application of wound healing assay to detect the regulation of ART on pancreatic cancer cell migration.10.Application of transwell invasion assay to detect the regulation of ART on pancreatic cancer cell invasion.11.Immunofluorescence was used to detect the epithelial-mesenchymal transition(EMT)in pancreatic cancer cells.12.Serine threonine phosphatase assay was used to analyze the activity of protein phosphatase 2A(PP2A).13.Real-time PCR was used to detect the mRNA expression of pro-angiogenic genes in pancreatic cancer cells.Results: 1.ART repressed the growth of pancreatic cancer cells in vitro and in vivo.2.ART inhibited metastasis of pancreatic cancer cells.3.ART promoted vascularization and upregulation of pro-angiogenic genes.4.ART upregulated pro-angiogenic genes by activating multiple kinase pathways.5.ART repressed the expression and activity of the protein phosphatase 2A(PP2A).6.ART upregulated pro-angiogenic genes through PP2Ac/NF-κB and PP2Ac/JNK-dependent manners.Conclusion: We verified that ART can repress the growth and metastasis of pancreatic cancer cells.Our present results indicated for the first time that ART can repress the expression and activity of the PP2 Ac.Moreover,ART can upregulate pro-angiogenic genes through PP2Ac/NF-κB-and PP2Ac/JNK-dependent manners,then promote the formation of tumor neovascularization.This potential unfavorable side effect could be blocked by bevacizumab,a widely clinically used anti-angiogenesis antibody.Our findings may shed a new theory for clinical practice of ART application to treating pancreatic cancer.