EZH2通过调控RAI2的表达影响肺腺癌细胞的恶性进展和DNA损伤修复

IF 1.5 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis Pub Date : 2022-07-01 DOI:10.1016/j.mrfmmm.2022.111792

Mingjiang Huang, Jianyang Ding, Xuhui Wu, Xuyang Peng, Gongzhi Wu, Congxiong Peng, Huaizhong Zhang, Chaofan Mao, Bin Huang

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引用次数: 1

摘要

肺腺癌(LUAD)具有高发病率和高死亡率的特点。组蛋白甲基转移酶EZH2的异常激活与癌症进展密切相关。本研究旨在深入探讨EZH2及其下游基因在LUAD细胞恶性进展及DNA损伤修复中的作用及可能的分子机制。方法采用qRT-PCR分析EZH2在LUAD细胞中的表达，采用CCK-8、集落形成和流式细胞术检测EZH2对LUAD细胞增殖和凋亡的影响。通过生物信息学分析预测了EZH2的下游靶点。然后，通过CHIP和荧光素酶报告基因检测EZH2和RAI2的靶向关系。通过挽救实验进一步验证EZH2/RAI2对LUAD细胞恶性进展的影响。Western blot检测EZH2、RAI2、p53的表达水平。结果在LUAD组织和细胞中发现EZH2的调控作用。RAI2是EZH2的下游靶基因，两者呈负相关。沉默EZH2抑制LUAD细胞的增殖，促进p53的表达，细胞周期阻滞和凋亡。而对RAI2的沉默可以逆转EZH2沉默引起的上述影响。结论EZH2通过靶向并负调控RAI2促进LUAD细胞恶性进展和DNA损伤修复。

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EZH2 affects malignant progression and DNA damage repair of lung adenocarcinoma cells by regulating RAI2 expression

Background

Lung adenocarcinoma (LUAD) is featured in high morbidity and mortality. Aberrant activation of the histone methyltransferase EZH2 has close association with cancer progression. This research aimed to deeply dive into the role and possible molecular mechanisms of EZH2 and its downstream genes in malignant progression and DNA damage repair of LUAD cells.

Methods

Expression of EZH2 in LUAD cells was analyzed by qRT-PCR, and the effects of EZH2 on proliferation, and apoptosis of LUAD cells were examined by CCK-8, colony formation and flow cytometry assays. The downstream targets of EZH2 were predicted by bioinformatics analysis. Then, the targeting relationship between EZH2 and RAI2 was examined by CHIP and luciferase reporter assays. Rescue assay were used to further validate the effect of EZH2/RAI2 on the malignant progression of LUAD cells. The expression levels of EZH2, RAI2 and p53 were examined by Western blot.

Results

Upregulation of EZH2 was identified in LUAD tissues and cells. RAI2 was a downstream target gene of EZH2, and the two were negatively correlated. Silencing EZH2 suppressed proliferation of LUAD cells, promoted expression of p53, cell cycle arrest and apoptosis. While silencing RAI2 could reverse the above-mentioned effects caused by EZH2 silencing.

Conclusion

These results demonstrated that EZH2 promoted malignant progression and DNA damage repair of LUAD cells by targeting and negatively regulating RAI2.

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来源期刊

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis 生物-毒理学

CiteScore

4.90

自引率

0.00%

发文量

审稿时长

51 days

期刊介绍： Mutation Research (MR) provides a platform for publishing all aspects of DNA mutations and epimutations, from basic evolutionary aspects to translational applications in genetic and epigenetic diagnostics and therapy. Mutations are defined as all possible alterations in DNA sequence and sequence organization, from point mutations to genome structural variation, chromosomal aberrations and aneuploidy. Epimutations are defined as alterations in the epigenome, i.e., changes in DNA methylation, histone modification and small regulatory RNAs. MR publishes articles in the following areas: Of special interest are basic mechanisms through which DNA damage and mutations impact development and differentiation, stem cell biology and cell fate in general, including various forms of cell death and cellular senescence. The study of genome instability in human molecular epidemiology and in relation to complex phenotypes, such as human disease, is considered a growing area of importance. Mechanisms of (epi)mutation induction, for example, during DNA repair, replication or recombination; novel methods of (epi)mutation detection, with a focus on ultra-high-throughput sequencing. Landscape of somatic mutations and epimutations in cancer and aging. Role of de novo mutations in human disease and aging; mutations in population genomics. Interactions between mutations and epimutations. The role of epimutations in chromatin structure and function. Mitochondrial DNA mutations and their consequences in terms of human disease and aging. Novel ways to generate mutations and epimutations in cell lines and animal models.