EP300 promotes bladder tumor migration and invasion by upregulating FKBP10

doi:10.3877/cma.j.issn.2095-1221.2024.05.002

Abstract

Abstract:

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'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'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.

Key words: Bladder tumor, EP300, FKBP10, Migration, Invasion

Xupeng Zhao, Jichen Wang, Shuo Tian, Hongzhao Li, Xiubin Li, Xu Zhang. EP300 promotes bladder tumor migration and invasion by upregulating FKBP10[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(05): 264-274.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2024.05.002

https://zhxbygxbzz.cma-cmc.com.cn/EN/Y2024/V14/I05/264

Figures/Tables 17

References 25

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基因	序列（5'→3'）	预期扩增产物长度（bp）
FKBP10	上游TGCGGATGTGGTGGAAATCA	141
	下游CCGTAGTCATGCGAGGTGAA
EP300	上游AGCCAAGCGGCCTAAACTC	144
	下游TCACCACCATTGGTTAGTCCC
GAPDH（内参）	上游ATTCCATGGCACCGTCAAGGCTGA	156
	下游TTCTCCATGGTGGTGAAGACGCCA
FKBP10 ChIP-qPCR	上游ATGCCTTCCCTTGGAGTACAG	100
	下游TTCGAATTCTAGTCCCCTCCAG
GAPDH ChIP-qPCR（内参）	上游TACTAGCGGTTTTACGGGCG	166
	下游TCGAACAGGAGGAGCAGAGAGCGA

基因	序列（5'→3'）	预期扩增产物长度（bp）
FKBP10	上游TGCGGATGTGGTGGAAATCA	141
	下游CCGTAGTCATGCGAGGTGAA
EP300	上游AGCCAAGCGGCCTAAACTC	144
	下游TCACCACCATTGGTTAGTCCC
GAPDH（内参）	上游ATTCCATGGCACCGTCAAGGCTGA	156
	下游TTCTCCATGGTGGTGAAGACGCCA
FKBP10 ChIP-qPCR	上游ATGCCTTCCCTTGGAGTACAG	100
	下游TTCGAATTCTAGTCCCCTCCAG
GAPDH ChIP-qPCR（内参）	上游TACTAGCGGTTTTACGGGCG	166
	下游TCGAACAGGAGGAGCAGAGAGCGA

分组	实验次数	EP300 mRNA相对表达量	EP300蛋白相对表达量
Scramble	3	1.01±0.16	0.99±0.04
shEP300-1	3	0.14±0.02^a	0.32±0.02^a
shEP300-2	3	0.17±0.05^a	0.31±0.03^a
F值		79.862	53.874
P值		＜0.05	＜0.05

分组	实验次数	EP300 mRNA相对表达量	EP300蛋白相对表达量
Scramble	3	1.01±0.16	0.99±0.04
shEP300-1	3	0.14±0.02^a	0.32±0.02^a
shEP300-2	3	0.17±0.05^a	0.31±0.03^a
F值		79.862	53.874
P值		＜0.05	＜0.05

分组	实验次数	第3天OD值	相对迁移面积（%）	相对迁移细胞量（%）	相对侵袭细胞量（%）
Scramble	3	1.49±0.05	79.38±2.30	100.32±5.16	99.52±3.84
shEP300-1	3	1.16±0.06^a	33.55±2.87^a	52.16±2.07^a	33.07±7.14^a
shEP300-2	3	1.07±0.04^a	43.76±1.89^a	43.19±7.64^a	64.22±4.15^a
F值		61.322	30.407	16.241	13.133
P值		＜0.05	＜0.05	＜0.05	＜0.05

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