川北医学院学报2025，Vol.40Issue(1) ：6-12.DOI:10.3969/j.issn.1005-3697.2025.01.002

ZDHHC20对m5C阅读器YBX1的棕榈酰化修饰通过调节mRNA m5C甲基化促进结直肠癌进展

Palmitoylation of m5C reader YBX1 by ZDHHC20 promotes colorectal cancer progression through regulating mRNA m5C methylation

罗城 蒙长远 夏术森 黄珊 彭洪
川北医学院学报2025，Vol.40Issue(1) ：6-12.DOI:10.3969/j.issn.1005-3697.2025.01.002
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  • 万方数据

ZDHHC20对m5C阅读器YBX1的棕榈酰化修饰通过调节mRNA m5C甲基化促进结直肠癌进展

Palmitoylation of m5C reader YBX1 by ZDHHC20 promotes colorectal cancer progression through regulating mRNA m5C methylation

罗城 1蒙长远 2夏术森 2黄珊 3彭洪3
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作者信息

  • 1. 遂宁市中医院普外科,四川遂宁 629000
  • 2. 川北医学院附属医院胃肠外科
  • 3. 南充市中心医院肛肠外科,四川南充 637000
  • 折叠

摘要

目的:探讨棕榈酰基转移酶20(ZDHHC20)通过棕榈酰化修饰调节5-甲基胞嘧啶(m5C)阅读器Y-框结合蛋白1(YBX1)蛋白稳定性,并提高mRNA m5C修饰水平参与结直肠癌(CRC)发生发展的分子机制.方法:通过生物信息学方法分析ZDHHC20 mRNA和蛋白在CRC组织中的表达及ZDHHC20表达水平与CRC患者临床病理特征的关系.通过RT-qPCR检测CRC细胞中ZDHHC20的表达水平.CCK-8检测ZDHHC20对CRC细胞增殖的影响,Transwell实验用于检测ZDHHC20对细胞侵袭的影响.通过生物信息学数据库分析ZDHHC20与YBX1的相关性及YBX1蛋白的棕榈酰化修饰位点.酰基生物素交换法(ABE)富集棕榈酰化蛋白,Western blot实验检测CRC细胞中YBX1的棕榈酰化修饰.过表达ZDHHC20检测YBX1的棕榈酰化修饰水平,并用Western blot实验检测2-BP和放线菌酮处理后YBX1蛋白的稳定性,CCK-8和Transwell法检测YBX1对CRC细胞增殖和迁移的影响,LC-MS法检测YBX1对CRC细胞中m5C修饰水平的影响.结果:ZDHHC20 mRNA和蛋白在CRC组织和细胞中高表达(P＜0.05),且ZDHHC20的表达与年龄、肿瘤分期和淋巴结转移相关(P＜0.05).过表达ZDH-HC20可促进CRC细胞增殖和侵袭(P＜0.05).ZDHHC20与YBX1表达水平正相关(P＜0.05),且YBX1上存在多个潜在棕榈酰化修饰位点.YBX1是棕榈酰化修饰蛋白,ZDHHC20可提高YBX1蛋白棕榈酰化修饰水平及YBX1蛋白稳定性(P＜0.05).YBX1可促进CRC细胞增殖和侵袭,并提高CRC细胞的m5C修饰水平(P＜0.05).结论:ZDHHC20可通过棕榈酰化修饰增强YBX1蛋白稳定性,提高mRNA m5C修饰水平并促进CRC细胞增殖和侵袭.

Abstract

Objective:To explore the molecular mechanism that palmitoyl acyltransferase 20(ZDHHC20)participates in the pathogenesis of colorectal cancer(CRC)through Palmitoylation modification to regulate 5-methylcytosine(m5C)reader Y-box binding proteinl(YBX1)protein stability,and improve mRNA m5C modification level.Methods:The expression of ZDHHC20 mRNA and pro-tein in CRC was analyzed by bioinformatics.Further analysis of the association among ZDHHC20 expression levels and the clinicopatho-logical features of CRC patients was performed.The expression level of ZDHHC20 in CRC cells was verified using RT-qPCR.A CCK-8 assay was used to detect the effect of ZDHHC20 on the proliferation of CRC cells.Transwell was used for cell invasion assays.The rela-tionship between the expression of ZDHHC20 and YBX1 in CRC and the palmitoylation site of YBX1 was investigated by bioinformatics database analysis.Palmitoylation protein was enriched by ABE method and Westernblot experiment was used to detect YBX1 palmitoyla-tion modification in CRC.Overexpression of ZDHHC20 was used to detect the palmitoylation modification level of YBX1.The Western blot was used to measure the stability of YBX1 protein under 2-BP and actinomycin.The influence of YBX1 on the proliferation and mi-gration of CRC cells was measured by CCK-8 and Transwell.LC-MS was used to analyze the impact of YBX1 on m5C modification level in CRC cells.Results:An upregulation of ZDHHC20 mRNA and protein were observed in the sample CRC cells and tissues(P＜0.05).ZDHHC20 expression was correlated with age,tumor stage and lymph node metastasis(P＜0.05).Overexpression of ZDHHC20 contributes to proliferation and invasion in CRC cells(P＜0.05).The expression between ZDHHC20 and YBX1 were posi-tively correlated(P＜0.05),and YBX1 had multiple palmitoylation modification site.YBX1 was a palmitoylated protein and ZDHHC20 could increase YBX1 palmitoylation modification level and improve the stability of YBX1 protein(P＜0.05).YBX1 could enhance CRC cell proliferation and invasion.Also YBX1 could improve m5C modification level in CRC(P＜0.05).Conclusion:ZDHHC20 can improve the stability of YBX1 protein through Palmitoylation modification and mRNA m5C modification level,and enhance CRC cell proliferation and invasion.

关键词

结直肠癌/5-甲基胞嘧啶/Y-框结合蛋白1/棕榈酰基转移酶20/增殖/侵袭

Key words

Colorectal Cancer/5-methylcytosine/Y-box binding protein l/Palmitoyl acyltransferase 20/Proliferation/Invasion

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出版年

2025
川北医学院学报
川北医学院

川北医学院学报

影响因子：0.958
ISSN：1005-3697
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