CRISPR/Cas9-mediated Knockout of LYVE1 In Human Tongue Cancer Cells Reveals Transcriptomic Changes in Metastasis-associated Pathways.

IF 2.6 4区 医学 Q2 GENETICS & HEREDITY
Sini Karinen, Johanna Forero-Rodríguez, Annika Järvinen, Kari K Eklund, Goncalo Barreto, Abdelhakim Salem
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引用次数: 0

Abstract

Background/aim: Tongue squamous cell carcinoma (TSCC), a highly aggressive subtype of head and neck cancers, is characterized by frequent lymphatic metastasis and poor prognosis. Recently, we showed that lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1) is involved in TSCC progression, yet the underlying molecular mechanisms remain unclear.

Materials and methods: CRISPR/Cas9 gene editing was employed to generate LYVE1 knockout (KO) TSCC cell lines. Single-cell clones were isolated, screened, and validated through sequencing and Inference of CRISPR Edits (ICE) analysis and qRT-PCR. RNA sequencing was performed on LYVE1 KO and wild-type (WT) cells to identify differentially expressed genes (DEGs). Bioinformatic analyses, including Gene Ontology (GO) enrichment and protein-protein interaction (PPI) network mapping, were conducted to explore affected pathways. Finally, network topology was examined using NetworkAnalyzer and cytoHubba plugins.

Results: Transcriptomic analysis revealed significant down-regulation of pro-metastatic pathways, including epithelial-mesenchymal transition (EMT), extracellular matrix remodeling, and immune modulation. DEG analysis identified 263 genes, with key down-regulated targets such as WNT5A, TGFB2, and MMP2, and up-regulation of tumor-suppressive genes including PTGS2. GO and PPI analyses highlighted LYVE1's pivotal role in regulating cell adhesion, migration, and immune response.

Conclusion: LYVE1 KO reduces TSCC invasive potential by disrupting EMT and tumor-stroma interactions, aligning with previous experimental findings. These results suggest LYVE1 as a critical driver of metastasis, highlighting its potential as a therapeutic target.

CRISPR/ cas9介导的人舌癌细胞LYVE1基因敲除揭示了转移相关途径的转录组学变化
背景/目的:舌鳞癌(TSCC)是头颈部肿瘤中一种高度侵袭性的亚型,其特点是淋巴转移频繁,预后差。最近,我们发现淋巴管内皮透明质酸受体1 (LYVE-1)参与了TSCC的进展,但其潜在的分子机制尚不清楚。材料与方法:采用CRISPR/Cas9基因编辑技术制备LYVE1敲除(KO) TSCC细胞株。通过测序和推断CRISPR编辑(ICE)分析和qRT-PCR对单细胞克隆进行分离、筛选和验证。对LYVE1 KO和野生型(WT)细胞进行RNA测序以鉴定差异表达基因(DEGs)。通过生物信息学分析,包括基因本体(GO)富集和蛋白蛋白相互作用(PPI)网络定位,探索受影响的途径。最后,使用NetworkAnalyzer和cytoHubba插件检查网络拓扑。结果:转录组学分析显示,促转移途径显著下调,包括上皮-间质转化(EMT)、细胞外基质重塑和免疫调节。DEG分析共鉴定出263个基因,其中WNT5A、TGFB2、MMP2等关键下调靶点,PTGS2等肿瘤抑制基因上调。GO和PPI分析强调了LYVE1在调节细胞粘附、迁移和免疫反应中的关键作用。结论:LYVE1 KO通过破坏EMT和肿瘤间质相互作用降低TSCC侵袭潜力,与先前的实验结果一致。这些结果表明LYVE1是转移的关键驱动因素,突出了其作为治疗靶点的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Cancer Genomics & Proteomics
Cancer Genomics & Proteomics ONCOLOGY-GENETICS & HEREDITY
CiteScore
5.00
自引率
8.00%
发文量
51
期刊介绍: Cancer Genomics & Proteomics (CGP) is an international peer-reviewed journal designed to publish rapidly high quality articles and reviews on the application of genomic and proteomic technology to basic, experimental and clinical cancer research. In this site you may find information concerning the editorial board, editorial policy, issue contents, subscriptions, submission of manuscripts and advertising. The first issue of CGP circulated in January 2004. Cancer Genomics & Proteomics is a journal of the International Institute of Anticancer Research. From January 2013 CGP is converted to an online-only open access journal. Cancer Genomics & Proteomics supports (a) the aims and the research projects of the INTERNATIONAL INSTITUTE OF ANTICANCER RESEARCH and (b) the organization of the INTERNATIONAL CONFERENCES OF ANTICANCER RESEARCH.
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