Pancreatic cancer cell-derived exosomes promote nerve invasion by regulating the NGF/TrkA pathway

Abstract

Abstract Objective To investigate the effect of pancreatic cancer cell-derived exosomes on the function of neurogenic cells and the role of NGF/TrkA signaling pathway, and to study the related mechanisms. Methods The exosomes of Panc02 cells were extracted by polymer precipitation method, and identified by transmission electron microscopy, particle size analysis and Western blot. Exosomes were co-cultured with RSC cells and PC-12 cells. The proliferation ability of the cells was detected by CCK8 assay and plate cloning assay. The migration ability of the cells was detected by Transwell assay and cell scratch healing assay. RSC cells and PC-12 cells were co-cultured with exosomes or TrkA inhibitor GW441756, and the changes of NGF/TrkA expression levels in neurogenic cells were detected by Western blot assay, and then the changes of cell proliferation and migration ability were detected by relevant experiments. Western blot assay was used to detect changes in the expression levels of neuroinvasion-related proteins, epithelial mesenchymal transformation (EMT) related proteins and AKT/mTOR pathway proteins in neuro-derived cells. Results The exosomes of Panc02 cells of pancreatic cancer were extracted successfully. CCK8 assay and plate cloning assay showed that Panc02 exosomes significantly enhanced the proliferation ability of neurogenic cells, Transwell assay and cell scratch healing assay showed that Panc02 exosomes significantly enhanced the migration ability of neurogenic cells. Western blot analysis showed that Panc02 exosomes up-regulated NGF/TrkA expression, TrkA inhibitor GW441756 down-regulated NGF/TrkA expression and inhibited the proliferation and migration of neural cells, while Panc02 exosomes reversed the inhibitory effect of TrkA inhibitor GW441756. After co-culture with exosomes, neuro-invasion-related proteins were upregulated, EMT capacity was enhanced, and AKT/mTOR pathway was activated and phosphorylated. Conclusion Pancreatic cancer cell derived exosomes promote the proliferation, migration and invasion of neurogenic cells by up-regulating NGF/TrkA activated AKT/m-TOR pathway.