Changes in invasion and migration abilities of lung adenocarcinoma cells with down-regulated histone acetyltransferase Tip60 and their underlying mechanisms
Objective To observe the changes in invasion and migration abilities of lung adenocarcinoma cells with down-regulated histone acetyltransferase Tip60 and to investigate their underlying mechanisms.Methods The lung ade-nocarcinoma cell line A549 was divided into the Tip60 down-regulation group,negative control for down-regulation group,and control group.Cells in the Tip60 down-regulation group and the negative control for down-regulation group were trans-fected with Tip60-specific interfering siRNA and negative control siRNA,respectively,while cells in the control group were not transfected.The SETDB1 protein level in cells was detected using Western blotting,and the SETDB1 mRNA lev-el was measured by real-time fluorescent quantitative PCR.Cell proliferation was assessed using the CCK-8 assay,cell in-vasion capability was evaluated using the Transwell chamber assay,and cell migration ability was observed using the wound healing assay.The significant enrichment peaks of histone H3 lysine 9(H3K9)and lysine 27(H3K27)were identi-fied in the promoter region of SETDB1 in A549 lung adenocarcinoma cells by using the UCSC Browser feature of the Cis-trome DB database.Chromatin immunoprecipitation(ChIP)was then performed to detect the acetylation levels of H3K9 and H3K27 in the promoter region of the SETDB1 gene.Results The expression levels of SETDB1 protein and mRNA in the Tip60 down-regulation group were lower than those in the control group and the negative control for down-regulation group(all P<0.05).At 24,48,and 72 h of culture,the cell proliferation ability of the Tip60 down-regulation group was lower than those of the control group and the negative control for down-regulation group.The number of invasive cells was smaller in the Tip60 down-regulation group than in the control group and the negative control for down-regulation group.At 24 and 48 h of culture,the wound healing rate of the Tip60 down-regulation group was lower than those in the control group and the negative control for down-regulation group(all P<0.05).The acetylation levels of histone H3K9 and H3K27 in the SETDB1 promoter region were lower in the Tip60 down-regulation group than in the control group and the negative control for down-regulation group(all P<0.05).Conclusion Down-regulation of Tip60 can inhibit the invasion and migration abilities of lung adenocarcinoma cells,and the mechanism may be related to the regulation of acetylation levels of histone H3 in the SETDB1 promoter region.
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