Cui Yishuang, Wu Yanan, Zhang Mengshi, Zhu Yingze, Su Xin, Kong Wenyue, Zheng Xuan, Sun Guogui
School of Public Health, North China University of Science and Technology, Department of Hebei Key Laboratory of Medical-Industrial Integration Precision Medicine, North China University of Science and Technology Affiliated Hospital, Tangshan, Hebei, China.
Department of Hebei Key Laboratory of Medical-Industrial Integration Precision Medicine, North China University of Science and Technology Affiliated Hospital, Tangshan, Hebei, China.
Front Oncol. 2023 Oct 25;13:1240868. doi: 10.3389/fonc.2023.1240868. eCollection 2023.
To discern long non-coding RNAs (lncRNAs) with prognostic relevance in the context of lung squamous cell carcinoma (LUSC), we intend to predict target genes by leveraging The Cancer Genome Atlas (TCGA) repository. Subsequently, we aim to investigate the proliferative potential of critical lncRNAs within the LUSC milieu.
DESeq2 was employed to identify differentially expressed genes within the TCGA database. Following this, we utilized both univariate and multivariate Cox regression analyses to identify lncRNAs with prognostic relevance. Noteworthy lncRNAs were selected for validation in cell lines. The intracellular localization of these lncRNAs was ascertained through nucleocytoplasmic isolation experiments. Additionally, the impact of these lncRNAs on cellular proliferation, invasion, and migration capabilities was investigated using an Antisense oligonucleotides (ASO) knockdown system.
Multivariate Cox regression identified a total of 12 candidate genes, consisting of seven downregulated lncRNAs (BRE-AS1, CCL15-CCL14, DNMBP-AS1, LINC00482, LOC100129034, MIR22HG, PRR26) and five upregulated lncRNAs (FAM83A-AS1, LINC00628, LINC00923, LINC01341, LOC100130691). The target genes associated with these lncRNAs exhibit significant enrichment within diverse biological pathways, including metabolic processes, cancer pathways, MAPK signaling, PI3K-Akt signaling, protein binding, cellular components, cellular transformation, and other functional categories. Furthermore, nucleocytoplasmic fractionation experiments demonstrated that LINC00923 and LINC01341 are predominantly localized within the cellular nucleus. Subsequent investigations utilizing CCK-8 assays and colony formation assays revealed that the knockdown of LINC00923 and LINC01341 effectively suppressed the proliferation of H226 and H1703 cells. Additionally, transwell assays showed that knockdown of LINC00923 and LINC01341 significantly attenuated the invasive and migratory capacities of H226 and H1703 cells.
This study has identified 12 candidate lncRNA associated with prognostic implications, among which LINC00923 and LINC01341 exhibit potential as markers for the prediction of LUSC outcomes.
为了在肺鳞状细胞癌（LUSC）背景下识别具有预后相关性的长链非编码RNA（lncRNA），我们打算利用癌症基因组图谱（TCGA）数据库预测靶基因。随后，我们旨在研究LUSC环境中关键lncRNA的增殖潜力。
采用DESeq2在TCGA数据库中识别差异表达基因。在此之后，我们利用单变量和多变量Cox回归分析来识别具有预后相关性的lncRNA。选择值得注意的lncRNA在细胞系中进行验证。通过核质分离实验确定这些lncRNA的细胞内定位。此外，使用反义寡核苷酸（ASO）敲低系统研究这些lncRNA对细胞增殖、侵袭和迁移能力的影响。
多变量Cox回归共鉴定出12个候选基因，包括7个下调的lncRNA（BRE-AS1、CCL15-CCL14、DNMBP-AS1、LINC00482、LOC100129034、MIR22HG、PRR26）和5个上调的lncRNA（FAM83A-AS1、LINC00628、LINC00923、LINC01341、LOC100130691）。与这些lncRNA相关的靶基因在多种生物途径中表现出显著富集，包括代谢过程、癌症途径、MAPK信号传导、PI3K-Akt信号传导、蛋白质结合、细胞成分、细胞转化和其他功能类别。此外，核质分级分离实验表明，LINC00923和LINC01341主要定位于细胞核内。随后利用CCK-8测定法和集落形成测定法进行的研究表明，敲低LINC00923和LINC01341可有效抑制H226和H1703细胞的增殖。此外，Transwell测定法表明，敲低LINC00923和LINC01341可显著减弱H226和H1703细胞的侵袭和迁移能力。
本研究鉴定出12个与预后相关的候选lncRNA，其中LINC00923和LINC01341具有作为预测LUSC预后标志物的潜力。
