摘要
目的探讨乙酰基转移酶抑制剂C646靶向KAT3b通过Survivin蛋白乙酰化影响食管癌细胞上皮间质转化(epithelial mesenchymal transformation,EMT)、迁移和侵袭的机制。方法体外培养食管癌TE-1细胞,MTT法筛选C646最佳给药浓度;C646处理TE-1细胞为实验组(C646组),DMSO处理TE-1细胞为对照组(DMSO组);实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测KAT3b mRN表达量;免疫共沉淀法检测Survivin蛋白乙酰化;Western blot法检测KAT3b及EMT相关蛋白表达;MTT法检测细胞存活能力,细胞划痕实验观察迁移能力,Transwell实验检测侵袭能力。结果C646组中KAT3b mRNA和蛋白表达量下降,且Survivin蛋白乙酰化水平下降;C646组中E-cadherin蛋白表达上调,而N-cadherin、Snail和Slug蛋白表达下降。C646组中食管癌TE-1细胞存活、迁移和侵袭能力显著下降。结论C646可能通过下调KAT3b的表达降低Survivin蛋白乙酰化水平,进而抑制食管癌细胞EMT、存活及迁移侵袭能力。
Objective To investigate the mechanism of acetyltransferase inhibitor C646 targets KAT3b in regulating the epithelial mesenchymal transformation(EMT),migration and invasion via acetylation of Survivin protein in esophageal cancer.Methods TE-1 cells of esophageal cancer were cultured in vitro,MTT method was used to screen the optimal administration concentration of C646;TE-1 cells treated with C646 were the experimental group(C646group),and TE-1 cells treated with DMSO were the control group(DMSO group);The expression of KAT3b mRNA was measured by real-time quantitative polymerase chain reaction(RT-qPCR);Co-IP measured Survivin protein acetylation;the expression of KAT3b and EMT-related proteins was determined by Western blot;Cell survival ability was measured by MTT,migration ability was observed by scratch,invasion ability was observed by Transwell.Results The KAT3b mRNA and protein expression levels decreased,and the Survivin protein acetylation levels decreased in the C646group.The expression of E-cadherin protein was up-regulated in the C646group,while the N-cadherin,Snail,and Slug proteins were decreased.The survival,migration,and invasion ability of esophageal cancer TE-1 cell were significantly decreased in the C646group.Conclusion C646 may reduce the acetylation level of Survivin protein by down-regulating KAT3b expression,and subsequently inhibit the EMT,survival,migration and invasion ability of oesophageal cancer cells.
作者
郑竞雄
杨阳
孙光蕊
邢磊
白宇航
梁宗英
ZHENG Jingxiong;YANG Yang;SUN Guangrui(Department of Thoracic Surgery,The Affiliated Hospital of Chengde Medical College,Hebei 067000,China)
出处
《医学研究杂志》
2023年第7期82-86,共5页
Journal of Medical Research
基金
河北省医学科学研究重点课题计划项目(20210985)。
