EP300通过上调FKBP10促进膀胱肿瘤细胞迁移和侵袭

EP300 promotes bladder tumor migration and invasion by upregulating FKBP10

目的探究E1A结合蛋白p300(EP300)在膀胱肿瘤进展中的功能及其潜在的分子调控机制,以期为膀胱肿瘤的治疗策略提供新的视角。方法首先在膀胱肿瘤细胞UMUC3中敲低EP300后,通过CCK-8实验、划痕实验、Transwell实验检测细胞的增殖、迁移和侵袭功能变化。进一步通过转录组测序和数据分析,寻找下游功能执行分子,并通过RT-qPCR、Western blot、ChIP-qPCR等实验验证EP300对其表达的调控。两组间比较使用t检验,3组或以上的两两比较使用单因素ANOVA分析,组间两两比较采用Dunnett-t检验。结果与对照组相比,敲低EP300后能够抑制UMUC3细胞的增殖(1.49±0.05比1.16±0.06、1.07±0.04,P<0.05)、迁移(100.32±5.16比52.16±2.07、43.19±7.64,P<0.05)及侵袭功能(99.52±3.84比33.07±7.14、64.22±4.15,P<0.05)。转录组测序结果表明敲低EP300后FK506结合蛋白10(FKBP10)下调(1.02±0.11比0.18±0.04,P<0.05)。加入EP300的乙酰转移酶功能抑制剂C646后,FKBP10的表达水平也下降(0.99±0.07比0.44±0.09,P<0.05)。结合ChIP-qPCR结果证实,EP300通过促进FKBP10启动子区域的H3K27乙酰化(1.40±0.05比0.54±0.01,P<0.05),进而调控FKBP10的转录活性。结论本研究揭示了EP300通过上调FKBP10的表达,促进膀胱肿瘤进展的分子机制。

Objective This study aims to investigate the role of E1A binding protein p300(EP300)in bladder tumor progression and its underlying molecular mechanisms,with the goal of providing new insights for bladder cancer therapeutic strategies.Methods EP300 was knocked down in UMUC3 bladder tumor cells,followed by assessments of cell proliferation,migration,and invasion using CCK-8,wound healing,and Transwell assays.Transcriptome sequencing and data analysis were conducted to identify downstream effector molecules.The regulation of these molecules by EP300 was validated using RT-qPCR,Western blot,and ChIP-qPCR assays.Comparisons between two groups were conducted using the t-test.For comparisons among three or more groups,one-way ANOVA was employed,followed by Dunnett&apos;s t-test for post hoc pairwise comparisons.Results Compared with the control group,knockdown of EP300 significantly inhibited the proliferation[(1.49±0.05)vs(1.16±0.06),(1.07±0.04),P<0.05],migration[(100.32±5.16)vs(52.16±2.07),(43.19±7.64),P<0.05],and invasion[(99.52±3.84)vs(33.07±7.14),(64.22±4.15),P<0.05]of UMUC3 cells(P<0.05).Transcriptome sequencing revealed that FK506 binding protein 10(FKBP10)was significantly downregulated upon EP300 knockdown[(1.02±0.11)vs(0.18±0.04),P<0.05].The expression level of FKBP10 also decreased[(0.99±0.07)vs(0.44±0.09),P<0.05]after treatment with C646,an inhibitor of EP300&apos;s acetyltransferase activity.ChIP-qPCR results confirmed that EP300 promotes the transcriptional activity of FKBP10 by enhancing H3K27 acetylation in the promoter region of FKBP10[(1.40±0.05)vs(0.54±0.01),P<0.05].Conclusion This study revealed that EP300 promotes bladder tumor progression by upregulating FKBP10 expression.