The Role Of 14-3-3σ In The Migration Of A549 Cell Induced By Cigarette Smoking Extract

Objectives:To investigate the relationship between 14-3-3σexpression and cell migration in A549 induced by cigarette smoking extract(CSE);And to explore whether DNA methylation is causing the expression of 14-3-3σdecreased in A549 induced by cigarette smoking extract.Methods:A549 were exposed to CSE for 16 weeks to simulate the state of lung cancer patients to continue smoking (Current Smoking, CS),then develop with complete medium for 8 weeks to simulate the state of lung cancer patients to stop smoking (Ex-smoking,ES),at the same time A549 has been cultured in normal culture medium to simulate the the state of non-smokers with lung cancer(Non-smoking,NS)as the control group.To detection mRNA expression of 14-3-3σby RT-PCR.When cells were transfected with empty plasmid,14-3-3σplasmid,detection mRNA expression protein expression of 14-3-3a by RT-PCR,Western blot, and cell migration and transfer by transwell,wound-healing assay;To detection methylation of 14-3-3σby Methylmion Specific PCR.Results:mRNA expression of 14-3-3σin CS and ES are lower than NS,while no significant difference between CS and ES.CS and ES transfected with empty plasmid,mRNA and protein expression,migration and transfer did not change significantly compared with before;while CS and ES transfected with 14-3-3σplasmid,mRNA and protein expression significantly increased,the migration and transfer reduced.The methylation of 14-3-3σare imperfect in NS,CS and ES;However,the methylation of 14-3-3σare significantly enhanced in CS,ES compared with in NS. Conclusion:1.This is a close relationship between increased cell migration and decreased 14-3-3a expression in A549 induced by CSE.2.DNA methylation is causing the expression of 14-3-3σdecreased in A549 induced by cigarette smoking extract.