Experimental Study On The Proliferation And Invasion Ability Of Neutrophil Extracellular Traps In Pancreatic Cancer Cells

Objective The incidence rate of pancreatic cancer in all countries is high,and the overall growth rate is high.As a very poor prognosis of digestive tract cancer,the diagnosis and treatment of pancreatic cancer is very difficult.The growth of neutrophils and its neutrophil extracellular traps(NETs)have been reported to be associated with the proliferation and invasion of many tumors,but their effects on pancreatic cancer are not yet clear.The purpose of this study was to investigate the correlation between the absolute value of neutrophils and tumor markers,tumor stage TNM and histological differentiation,and the effect of NETs on the proliferation and invasion of pancreatic cancer cells(PANC-1)in order to provide new targets and ideas for the diagnosis and treatment of pancreatic cancer.Methods To collect the information of 20 cases of pancreatic cancer and 23 healthy persons who were in compliance with the standard of Department of general surgery in August 2019-2021 December,including the age,gender,blood routine,tumor series,TNM staging and histological differentiation,etc.Western blot(WB)was used to detect the effect of PANC-1 cells on the production of nets by neutrophils;CCK-8 experiment and plate clone formation experiment were used to detect the effect of nets on the proliferation of PANC-1 cells;The effect of nets on the invasive ability of PANC-1 cells was detected by invasion experiment.Results Clinical data showed that the white blood cell count,neutrophil absolute value and neutrophil percentage in patients with pancreatic cancer were higher than those in healthy subjects,with statistical significance(P < 0.05).The percentages of RBC,Hb and lymphocyte in patients with pancreatic cancer were lower than those in healthy subjects,with statistical significance(P < 0.05).There was no significant difference in age,gender,eosinophil percentage,basophil percentage and platelet count between the two groups of pancreatic cancer patients and healthy subjects,with no significant difference(P>0.05).The CA19-9 level and TNM stage of pancreatic cancer patients with high level of neutrophil absolute value were significantly higher than those of patients with absolute low neutrophils(P < 0.05).The level of tissue differentiation in patients with high level of neutrophil absolute value was lower than that in patients with pancreatic cancer whose absolute level of neutrophils was low.(P < 0.05),but there was no significant difference in CEA level between the two groups(P>0.05).PANC-1 cells co cultured with normal neutrophils could significantly stimulate the production of neutrophil elastase(NE),citrullinated histone H3(cit-h3)and myeloperoxidase(MPO),which was significantly different from the data of the control group(P < 0.05);CCK-8 experiment and plate clone formation experiment showed that nets could react on PANC-1 cells and significantly enhance their proliferation(P < 0.05);Invasion experiment showed that nets could enhance the invasion ability of PANC-1 cells,which was statistically significant(P < 0.05).Conclusions Pancreatic cancer patients had significantly higher levels of neutrophils than healthy individuals,and their levels increased with increasing CA19-9,TNM stage of the tumor,and malignancy of tissue differentiation;Pancreatic cancer cells are able to promote neutrophils to produce more nets;Nets can enhance the proliferation and invasion of pancreatic cancer cells.