P300低表达食管癌细胞株TE-1的Survivivn蛋白乙酰化、增殖迁移侵袭能力变化及其机制OACSTPCD
Changes of Survivivn protein acetylation,proliferation,migration and invasion of esophageal cancer TE-1 cell line with low expression of P300 and the mechanism
目的 观察乙酰基转移酶P300低表达食管癌细胞株TE-1细胞中的Survivivn蛋白乙酰化、增殖迁移侵袭能力变化,并探讨其机制.方法 取对数生长期TE-1细胞分为si-P300组、si-P300-NC组、NC组.si-P300组采用脂质体介导法转染乙酰基转移酶P300下调质粒,si-P300-NC组采用脂质体介导法转染阴性对照质粒,NC组不转染质粒,转染后继续孵育48 h进行后续实验.取各组细胞,采用免疫共沉淀(Co-IP)实验检测Survivin蛋白乙酰化水平.取各组细胞,继续培养12、24、36、48 h时,采用CCK-8法检测细胞增殖能力(以OD值表示).取各组细胞,采用细胞划痕实验检测迁移能力(以细胞迁移距离表示),采用Transwell小室实验检测侵袭能力(以穿膜细胞数表示),采用Western blotting法检测VEGF信号通路相关蛋白AKT1、MDM2、Bcl-2、Caspase3.结果 si-P300组细胞Survivin蛋白乙酰化水平均低于si-P300-NC组、NC组(P均<0.05).si-P300组细胞培养12、24、36、48 h时的OD值均低于si-P300-NC组、NC组(P均<0.05).si-P300组细胞迁移距离和穿膜细胞数均低于si-P300-NC组、NC组(P均<0.05).si-P300组细胞AKT1、MDM2、Bcl-2蛋白相对表达量均低于si-P300-NC组、NC组(P均<0.05),Caspase3蛋白相对表达量均高于si-P300-NC组、NC组(P均<0.05).结论 转染P300下调质粒抑制P300表达后,TE-1细胞乙酰化水平下降,细胞增殖、迁移、侵袭能力降低,其机制可能与调控VEGF信号通路有关.
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.
郑竞雄;孙光蕊;黄景涛;韩晓丽;赵松;杨悦;梁宗英
承德医学院附属医院胸外科,河北 承德 067000
乙酰化乙酰基转移酶P300Survivin蛋白食管癌VEGF信号通路
acetylationacetyltransferaseP300Survivin proteinesophageal carcinomaVEGF signaling pathway
《山东医药》 2024 (033)
15-18 / 4
河北省医学科学研究重点课题计划项目(20210985).
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