Benproperine Phosphate Inhibit Pancreatic Cancer Growth By Induce Autophagy

Objective Pancreatic cancer(PC)is one of the most common human malignancies worldwide.Currently,the most effective clinical treatment of PC is still surgical resection,which causes great suffering to patients especially the elderly.As such,there is an urgent need to develop novel therapeutic agents for efficient treatment of PC.Drug repurposing has reducing the time,cost and risk of drug development was compared with the development of new drugs,which have been successful in the medical field.Autophagy has been found to play a protective role when tumor cells are under nutrient shortage,hypoxia,and oxidative stress,and chemotherapyinduced cell death.In addition to protective autophagy,there are also type II programmed cell deaths that can cause cells to inhibit cell autophagy in tumors.Especially in the occurrence and development of pancreatic cancer,autophagy plays a key role in it.BPP is widely used as a non-productive cough suppressant It has been reported that BPP inhibits metastasis and angiogenesis of cancer cells.Together,our study identifies BPP as a potent anti-tumor agent for PC by inducing autophagy arrest,providing a new potential therapeutic strategy for the treatment of PC.Methods In this study,pancreatic cancer cell lines Panc-1 and MIA-Pa Ca-2 were selected as experimental objects,and observation and detection were performed at different concentrations.The effects of BPP on the growth and proliferation of pancreatic cancer cells were detected by MTT experiments,clone formation experiments,Ed U incorporation assay,experiments and LDH release cytotoxicity tests;Western Blotting,immunofluorescence,transfection of GFP-LC3 B,transfection of GFP-RFP tagged LC3 B plasmids,in the case of BPP or combined autophagy inhibitors(CQ and 3-MA)treatment and RNA interference(si RNA),Observe the changes of autophagy related proteins(LC3,Atg5,Beclin1)and autophagy flow;Acridine orange staining and Lyso-Tracker Red staining were used to observe the effect of BPP drugs on lysosomal acidity.After transient transfection of RAB11 A,Western Blotting and immunofluorescence experiments were used to observe the changes of autophagy-related proteins;BPP combined with autophagy inhibitors was detected by MTT,clone formation,Ed U incorporation assay and LDH release to detect the effect of autophagy on pancreatic cancer cells in the case of BPP drug treatment.In vivo experiments,a pancreatic cancer cell line Panc-1 was used to establish a pancreatic cancer subcutaneous tumor transplantation model.Mice were administrated with BPP drugs for 21 days to observe the size,volume and weight of pancreatic cancer xenografts.The changes of Ki67,LC3 and RAB11 A protein levels were detected by immunohistochemistry.Results BPP inhibits cell growth of human pancreatic cancer in vitro and in vivo.BPP induces autophagy in pancreatic cancer cells.BPP blocks autophagic flux in pancreatic cancer cells.BPP blocks autophagic flux by down-regulating RAB11 A in PC cells.BPP induces cytotoxicity autophagy in PC cells.Conclusions Here,we found that BPP showed significant anti-cancer effect on PC both in vitro and in vivo via induction of autophagy arrest.Mechanistical studies revealed that BPP reduced the expression of RAB11 A,disturbing the fusion of autophagosomes with lysosomes,thus leading to excessive accumulation of autophagosomes.Overexpression of RAB11 A or inhibition of autophagy partially reversed BPP-induced growth inhibition in PC cells,suggesting that BPP induced lethal autophagy arrest in PC cells.Together,our study identifies BPP as a potent anti-tumor agent for PC by inducing autophagy arrest,providing a new potential therapeutic strategy for the treatment of PC.