The Effect And Mechanism Of Panax Notoginseng Saponins Inhibit The Growth Of Pancreatic Cancer Cells And Improve Their Chemosensitivity Through The Autophagy And Apoptosis Pathways

Objective: Panax Notoginseng Saponins(PNS)is the main chemical composition of the Chinese herbal medicine Panax notoginseng,which has the functions of reducing vascular resistance and blood viscosity,improving microcirculation,inhibiting inflammatory factors and anti-oxidative stress.It has extensive pharmacological activities in the treatment of cardiovascular and cerebrovascular diseases and neural system diseases.Recently,PNS has received great attention and become the focus in the anti-tumor alternative medicine due to its potential antitumor activity and low toxicity.Studies have proven that PNS has significant anticancer effects on breast cancer,colorectal cancer,non-small cell lung cancer and cervical cancer,but the role of PNS in pancreatic cancer is rarely reported.In this study,we combined PNS and Gemcitabine(Gem)on pancreatic cancer cells to observe the phenotypic changes of pancreatic cancer cells,aiming to study whether PNS has anti-cancer effects on pancreatic cancer cells and the possible mechanism.Methods: Pancreatic cancer Miapaca2 and PANC-1 cells were treated with PNS and Gem with different drug concentration gradients,respectively.The viability of Miapaca2 and PANC-1 cells were detected by CCK-8 assay,and the IC50 values of PNS and Gem were calculated.The drug concentration and intervention time of PNS and Gem in the follow-up experiments were determined according to the results of the CCK-8 assay.The pancreatic cancer cells were divided into four groups: Control group,PNS group,Gem group and PNS + Gem group.The cells treated with the same amount of PBS were used as the control.The propagation of pancreatic cancer cells was tested through plate cloning experiment and Ed U cell proliferation experiment.Wound healing assay and Transwell assay were respectively used to detect the migration and invasion ability of pancreatic cancer cells.The apoptosis rate of pancreatic cancer cells was detected by flow cytometry.The expression levels of invasion,metastasis,apoptosis,and autophagy-related proteins MMP2,MMP9,cleaved-caspase3,Bax,Bcl-2,LC3B-I,LC3B-II,and p62 were quantitatively analyzed by Western Blot.Results: PNS inhibited the viability of pancreatic cancer Miapaca2 and PANC-1 cells in a dose-dependent and time-dependent manner,and the Miapaca2 cell line was more sensitive to PNS.In the plate cloning experiment,the formation of Miapaca2 and Panc-1 cell colonies was significantly inhibited in the PNS group,and the inhibitory effect was the largest in the PNS + Gem group.We also obtained the same trend in Ed U cell proliferation experiment,confirming that PNS can inhibit the proliferation of pancreatic cancer cells and improve the chemosensitivity of pancreatic cancer cells to Gem.Wound healing experiment and Transwell experiment showed that the migration and invasion ability of Miapaca2 and PANC-1 cells in PNS group were observably suppressed.Flow cytometry showed that PNS increased the apoptosis rate of Miapaca2 and PANC-1 cells.Western blot analysis showed that the expression of MMP2 and MMP9 proteins,which are closely related to cell migration and invasion,as well as the anti-apoptotic protein Bcl-2 were decreased,and the expression of pro-apoptotic proteins cleaved-caspase3 and Bax were increased in the PNS group.In addition,the ratio of autophagy-related protein LC3B-II/LC3B-I in Miapaca2 and PANC-1 cells was decreased and the expression of p62 protein was increased in the PNS group.Conclusions: PNS can effectively inhibit the proliferation,migration and invasion of pancreatic cancer cells,which may be achieved by inhibiting autophagy and inducing apoptosis.The combination of PNS and Gem has a stronger anticancer effect on pancreatic cancer cells,demonstrating that PNS can improve the chemosensitivity of pancreatic cancer cells to Gem.