In Vivo Study About The Effect Of Cigarette Smoke On The Biological Functions Of Neutrophils In Tumor Microenvironment

Objective: The purpose of this study was to study the effect of CSE on the biological function of neutrophilic granulocytes induced by tumor supernatant fluid.Methods: Neutrophils were extracted from healthy human blood.Tumor conditioned medium and cigarette smoke filtrate was prepared.CCK-8 method was used to determine safe CSE experimental concentration.Changes in the biological functions of the neutrophils,after 3 h and 24 h of culture in presence of CSE and tumor supernatants,were evaluated through measurements of cell viability,ROS intensity,adhesion and protein expression.Neutrophils alone were used as blank control and those induced by tumor supernatant were set as negative control.Results: 1.The activity of neutrophil after 3 h and 24 h of culture showed the same tendency.With the increase of CSE concentration,the relative survival rate of neutrophils showed a decreasing trend.2.CSE promoted the proliferation of human bladder cancer T24 cells in low concentration(p<0.05).While the cell survival rate of human bladder cancer T24 cells after 24 h of co-culture with high concentrations of CSE was significantly decreased(p<0.01).3.The viability of neutrophils could be enhanced by incubation with CSE,which was higher than that observed in blank control and negative control.4.The ability of adhesion of TCM-treated neutrophils to human umbilical vein endothelial HUVEC cells was significantly reduced after incubation with CSE.5.Reactive oxygen species(ROS)produced by neutrophils induced with tumor conditioned medium was significantly reduced by cigarette smoke filtrate.6.The expression of BV8 protein in TCM-induced neutrophils was up-regulated by cigarette smoke filtrate,while the protein expression of TNF-? and ICAM-1 was down-regulated.Conclusion: Cigarette smoke extracts can affect the biological function of neutrophils in tumor microenvironment,including increase of viability,reduction of adhesion,decrease in the expression of ROS and tumor-associated protein.In addition,cigarette smoke extracts could induce the differentiation of neutrophils to N2 type.