Effects Of Nicotine On Cell Proliferation,Migration And And Invasion Of Human A549

Objective To study the effects of nicotine on cell proliferation,invasion and migration of A549 andexplore the possible mechanisms.Methods A549 cells were cultured with different concentrations of nicotine to detect cell proliferation ability by CCK-8.The abilities of migration and invasion of A549 cell were measured by CCK-8,Transwell test,and wound scratch assay respectively.The expression of?7 nAChR,VEGF and MMP-2 was observed by western blotting after nicotine treatment.Results1.The cell viability of A549 cells treated with different concentrations of nicotine?10-8?10-3M?was gradually increased,and then decreased after reaching the peak.Cell viability in the nicotine concentration of 10-6M?10-5M range increased,compared with solvent control group.The cell viability reached a peak at 10-6M.The viability increased in A549 cells induced by 10^-6M of nicotine treatment,compared with solvent control group,the difference was statistically significant?P<0.05?.The results indicated that nicotine could promote the proliferation of human A549 cells.2.In the Transwell invasion assay,the number of A549 cells passing through Matrigel matrix was 29±4 in the blank control group and 56±7in the nicotine group.the difference was statistically significant?P<0.05?.The results suggested that nicotine could promote A549 cell invasion in vitro.3.The cell migration rate was?7.43±3.46?%in the blank control group and?46.35±9.77?%in the nicotine treatment group at 24 h after scratches.Wound-healing assay showed that nicotine could significantly promote the migration of A549 cells,the difference was statistically significant?P<0.05?.4.Western blotting analysis was used to detect the expressions of?7nAChR?VEGF and MMP-2 induced by nicotine treatment for 24h.The results indicated that increased expressions of a7-nAChR protein,VEGF protein and MMP-2 protein were observed in A549 cells induced by10^-6M of nicotine treatment for 24h,compared with solvent control group.the difference was statistically significant?P<0.05?.Conclusion:Nicotine increased proliferation,migration and invasion of human lung adenocarcinoma A549 cell through?7 nAChR,which may be associated with the up-regulation expression of VEGF and MMP-2protein.