Changes of Survivivn protein acetylation,proliferation,migration and invasion of esophageal cancer TE-1 cell line with low expression of P300 and the mechanism
Objective To observe the changes of Survivivn protein acetylation,proliferation,migration and inva-sion of esophageal cancer cell line TE-1 with low expression of acetyltransferase P300,and to explore the mechanism.Methods TE-1 cells in the logarithmic growth phase were divided into the si-P300 group,SI-P300-NC group and NC group,respectively.TE-1 cells in the Si-P300 group were transfected with acetyl transferase P300 down-regulated plasmid by liposome mediated method,TE-1 cells in the SI-P300-NC group were transfected with negative control plasmid by lipo-some mediated method,and TE-1 cells in the NC group were transfected without plasmid and were incubated for 48 h after transfection for follow-up experiment.Cells from each group were collected and Survivin protein acetylation level was de-tected by Co-IP assay.Cells were taken from each group and cultured for 12,24,36 and 48 h.The cell proliferation abili-ty(expressed by OD value)was detected by CCK-8.Cells in each group were selected,and the migration ability(repre-sented by cell migration distance)was detected by cell Scratch test,and the invasion ability(represented by the number of transmembrane cells)was detected by Transwell chamber test.VEGF signaling pathway-related proteins AKT1,MDM2,Bcl-2,and Caspase3 were detected by Western blotting.Results The acetylation level of Survivin protein was lower in the si-P300 group than in the si-P300-NC group and NC group(both P<0.05).The OD values of the si-P300 group were lower than those of the si-P300-NC group and NC group at 12,24,36 and 48 h after cell culture(all P<0.05).The cell migration distance was shorter and the number of transmembrane cells was smaller in the si-P300 group than in the si-P300-NC group and NC group(all P<0.05).The relative expression levels of AKT1,MDM2 and Bcl-2 protein in the si-P300 group were lower than those in the si-P300-NC group and NC group(all P<0.05),and the relative expression level of Cas-pase3 protein was higher than those in the si-P300-NC group and NC group(all P<0.05).Conclusion After transfec-tion with P300 down-regulated plasmid inhibits the expression of P300,the acetylation level of TE-1 cells decreases,and the cell proliferation,migration and invasion decrease,which may be related to the regulation of VEGF signaling pathway.
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