DNA methylation-regulated DDX27 promotes colorectal cancer progression through EZH2

IF 3.2 4区医学 Q2 PATHOLOGY

Pathology, research and practice Pub Date : 2025-10-01 DOI:10.1016/j.prp.2025.156247

Qin Chen , Jiawei Song , Binbin Zhang, Hongyu Wang, Juan Xu, Xiuqin Li, Xuelian Wang

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引用次数: 0

Abstract

Objective

To investigate the oncogenic role of DDX27 in colorectal cancer (CRC) and its regulation of EZH2.

Methods

DDX27 expression and prognostic value were analyzed using TCGA. qRT-PCR was performed on 30 paired CRC and adjacent tissues; DDX27 levels in CRC cell lines (HCT116, HT-29, SW480, SW620, HCT-8) were assessed by qRT-PCR and Western blot. Methylation-specific PCR evaluated CpG methylation of the DDX27 promoter and the effect of 5-azacytidine (5-AZ). Functional assays (CCK-8, flow cytometry, wound healing, transwell) examined proliferation, apoptosis, migration and invasion after DDX27 knockdown (si-DDX27). RNA-seq and bioinformatics analyses, together with Western blot assays, were used to identify downstream effectors. Luciferase reporter assays tested EZH2 promoter activity following DDX27 depletion. Rescue experiments overexpressed EZH2 in DDX27-silenced cells. In vivo, HCT116 cells with DDX27 shRNA or control shRNA were implanted in mice and tumor EZH2 was evaluated by IHC.

Results

DDX27 was upregulated in CRC and correlated with poorer prognosis. Promoter hypomethylation was observed and 5-AZ increased DDX27 expression. DDX27 knockdown inhibited proliferation, migration and invasion and increased apoptosis. Silencing DDX27 reduced EZH2 protein and significantly lowered EZH2 promoter luciferase activity. EZH2 overexpression partially rescued the anti-tumor effects of DDX27 silencing. In vivo DDX27 depletion suppressed tumor growth and reduced EZH2 expression.

Conclusions

DDX27 promotes CRC progression partly via transcriptional upregulation of EZH2; promoter hypomethylation contributes to DDX27 overexpression. Targeting the DDX27-EZH2 axis may be therapeutically valuable.

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DNA甲基化调控的DDX27通过EZH2促进结直肠癌的进展

目的探讨DDX27在结直肠癌（CRC）中的致瘤作用及其对EZH2的调控作用。方法采用TCGA分析ddx27的表达及预后价值。对30对结直肠癌及其邻近组织进行qRT-PCR检测；采用qRT-PCR和Western blot检测结直肠癌细胞株（HCT116、HT-29、SW480、SW620、HCT-8）中DDX27的表达水平。甲基化特异性PCR评估DDX27启动子的CpG甲基化和5-氮杂胞苷（5-AZ）的作用。功能检测（CCK-8、流式细胞术、伤口愈合、transwell）检测DDX27敲低后的增殖、凋亡、迁移和侵袭。RNA-seq和生物信息学分析以及Western blot检测用于鉴定下游效应物。荧光素酶报告基因检测检测DDX27耗竭后EZH2启动子活性。援救实验中，ddx27沉默细胞过表达EZH2。体内植入DDX27 shRNA或对照shRNA的HCT116细胞，通过免疫组化评价肿瘤EZH2。结果ddx27在结直肠癌中表达上调，与预后不良相关。观察到启动子低甲基化，5-AZ增加DDX27的表达。敲低DDX27抑制细胞增殖、迁移和侵袭，增加细胞凋亡。沉默DDX27可降低EZH2蛋白，显著降低EZH2启动子荧光素酶活性。EZH2过表达部分恢复了DDX27沉默的抗肿瘤作用。体内DDX27缺失抑制肿瘤生长，降低EZH2表达。结论ddx27部分通过上调EZH2转录促进结直肠癌进展；启动子低甲基化导致DDX27过表达。靶向DDX27-EZH2轴可能具有治疗价值。

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来源期刊

Pathology, research and practice 医学-病理学

CiteScore

5.00

自引率

3.60%

发文量

405

审稿时长

24 days

期刊介绍： Pathology, Research and Practice provides accessible coverage of the most recent developments across the entire field of pathology: Reviews focus on recent progress in pathology, while Comments look at interesting current problems and at hypotheses for future developments in pathology. Original Papers present novel findings on all aspects of general, anatomic and molecular pathology. Rapid Communications inform readers on preliminary findings that may be relevant for further studies and need to be communicated quickly. Teaching Cases look at new aspects or special diagnostic problems of diseases and at case reports relevant for the pathologist''s practice.