The Related Studies Of Nm23-H1 Gene Expression And Enhanced Invasion Of Human Lung Adenocarcinoma Cell Treated By Cigarette Smoking Extract

Objectives:To investigate the metastasis suppressor gene nm23-H1 on invasion ability of human lung adenocarcinoma cell line A549 with cigarette smoking extract (CSE) treatment;and to explore the mechanism of nm23-H1 down-regulated.Methods:The human lung adenocarcinoma cell line A549 was treated with 1%CSE for 16 weeks, the treated cells continued to culture for 8 weeks without CSE.Above both treated cells were transfected with nm23-H1 wild vector,while setting empty vector transfected group and untransfected group as the contro1,RT-PCR and western blot were used to detect nm23-H1 mRNA transcription and protein expression levels in six groups;using transwell chamber and wound-healing assay to test cell invasion and migration ability;methylation-specific PCR(MSP) was performed to examine the methylation status of nm23-H1 gene in A549 and both treated cells.Results:RT-PCR and western blot results showed nm23-H1 mRNA transcription and protein expression levels were up-regulated in nm23-H1 plasmid transfected cells and there were different significance compared with the others;transwell chamber and wound-healing assay also tested that cell invasion and migration ability was decreased in nm23-H1 plasmid transfected cells while there were different significance compared with the others.MSP showed that methylation level of nm23-H1 promoter was increased in A549 after treated with CSE.Conclusions:The increased invasion and migration ability of the treated-cells was probably related to nm23-H1 expression low-level;while nm23-H1 expression low-level was probably related to increased methylation level of nm23-H1 promoter.