CRISPR-Cas9 Mediated AGO2 Knockout inhibits tumorigenesis in human colorectal cancer cells.

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-07-28 DOI:10.14715/cmb/2024.70.7.25
Yizhi Zhu, Gengfang Wang, Haoran Xu, Yuan Guo
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Abstract

AGO2 plays a vital role in small RNA-guided gene silencing, which has been implied in the tumorigenesis of different types of tumors. Fundamentally, increased expression of AGO2 protein is associated with cancer progression and metastasis. This study aims to investigate the molecular mechanism by which AGO2 promotes tumorigenesis in colorectal cancer (CRC). Databases were used to analyze the expression levels of AGO2 in CRC and confirmed by a quantitative reverse transcriptase-PCR (qRT-PCR) assay in CRC tissues and normal adjacent tissues collected from 25 CRC patients. CRISPR/Cas9-mediated genome editing was used to knockout the AGO2 in HCT116 cells as a model system for colorectal cancers. The cell proliferation, migration and invasion ability of HCT116 cells were detected by CCK-8 assay, Wound scratch assay and Transwell assay. Moreover, the quantities of miRNA binding with AGO2 were detected by RNA-Binding Protein Immunoprecipitation (RIP-Assay). We demonstrated that AGO2 was aberrantly high-expressed in 25 matched-tissue pairs of colorectal cancer and para-carcinoma tissue. The following functional experiments verified that knockout of AGO2 suppressed cell proliferation, migration and tumorigenesis to hamper the aggressiveness of CRC. Our study also suggests a possible link between AGO2 and miRNA in RISC. AGO2 was elevated in CRC and knockout of AGO2 suppressed proliferation and tumorigenicity of CRC cells. Moreover, RISC formation and the function of miRNAs are also subject to AGO2. AGO2 may be a meaningful target for CRC therapy.

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CRISPR-Cas9 介导的 AGO2 基因敲除抑制了人类结直肠癌细胞的肿瘤发生。
AGO2 在小 RNA 引导的基因沉默中发挥着至关重要的作用,这在不同类型的肿瘤发生过程中都有暗示。从根本上说,AGO2 蛋白表达的增加与癌症的进展和转移有关。本研究旨在探讨 AGO2 促进结直肠癌(CRC)肿瘤发生的分子机制。研究利用数据库分析了 AGO2 在 CRC 中的表达水平,并通过定量反转录酶-PCR(qRT-PCR)检测证实了 AGO2 在 25 例 CRC 患者的 CRC 组织和正常邻近组织中的表达水平。利用 CRISPR/Cas9 介导的基因组编辑技术敲除了作为结直肠癌模型系统的 HCT116 细胞中的 AGO2。通过 CCK-8 试验、伤口划痕试验和 Transwell 试验检测了 HCT116 细胞的增殖、迁移和侵袭能力。此外,我们还利用 RNA 结合蛋白免疫沉淀(RIP-Assay)技术检测了 miRNA 与 AGO2 结合的数量。我们证实,在 25 对匹配的结直肠癌和癌旁组织中,AGO2 存在异常高表达。随后的功能实验证实,敲除 AGO2 可抑制细胞增殖、迁移和肿瘤发生,从而抑制 CRC 的侵袭性。我们的研究还表明,AGO2 与 RISC 中的 miRNA 之间可能存在联系。AGO2 在 CRC 中升高,敲除 AGO2 可抑制 CRC 细胞的增殖和致瘤性。此外,RISC 的形成和 miRNA 的功能也受 AGO2 的影响。AGO2可能是治疗CRC的一个有意义的靶点。
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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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