通过高通量测序鉴定低氧诱导的 A549 肺癌细胞中的关键 lncRNAs 和 mRNAs 并研究其潜在机制。

Identifying crucial lncRNAs and mRNAs in hypoxia-induced A549 lung cancer cells and investigating their underlying mechanisms via high-throughput sequencing.

机构信息

Department of Respiratory Medicine, The Second Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang, People's Republic of China.

Department of Oncology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, People's Republic of China.

出版信息

PLoS One. 2024 Sep 5;19(9):e0307954. doi: 10.1371/journal.pone.0307954. eCollection 2024.

DOI:10.1371/journal.pone.0307954

PMID:39236027

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11376552/

Abstract

BACKGROUND

Rapid proliferation and outgrowth of tumor cells frequently result in localized hypoxia, which has been implicated in the progression of lung cancer. The present study aimed to identify key long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) involved in hypoxia-induced A549 lung cancer cells, and to investigate their potential underlying mechanisms of action.

METHODS

High-throughput sequencing was utilized to obtain the expression profiles of lncRNA and mRNA in both hypoxia-induced and normoxia A549 lung cancer cells. Subsequently, a bioinformatics analysis was conducted on the differentially expressed molecules, encompassing functional enrichment analysis, protein-protein interaction (PPI) network analysis, and competitive endogenous RNA (ceRNA) analysis. Finally, the alterations in the expression of key lncRNAs and mRNAs were validated using real-time quantitative PCR (qPCR).

RESULTS

In the study, 1155 mRNAs and 215 lncRNAs were identified as differentially expressed between the hypoxia group and the normoxia group. Functional enrichment analysis revealed that the differentially expressed mRNAs were significantly enriched in various pathways, including the p53 signaling pathway, DNA replication, and the cell cycle. Additionally, key lncRNA-miRNA-mRNA relationships, such as RP11-58O9.2-hsa-miR-6749-3p-XRCC2 and SNAP25-AS1-hsa-miR-6749-3p-TENM4, were identified. Notably, the qPCR assay demonstrated that the expression of SNAP25-AS1, RP11-58O9.2, TENM4, and XRCC2 was downregulated in the hypoxia group compared to the normoxia group. Conversely, the expression of LINC01164, VLDLR-AS1, RP11-14I17.2, and CDKN1A was upregulated.

CONCLUSION

Our findings suggest a potential involvement of SNAP25-AS1, RP11-58O9.2, TENM4, XRCC2, LINC01164, VLDLR-AS1, RP11-14I17.2, and CDKN1A in the development of hypoxia-induced lung cancer. These key lncRNAs and mRNAs exert their functions through diverse mechanisms, including the competitive endogenous RNA (ceRNA) pathway.

摘要

背景

肿瘤细胞的快速增殖和生长常导致局部缺氧，这与肺癌的进展有关。本研究旨在鉴定参与缺氧诱导的 A549 肺癌细胞的关键长链非编码 RNA（lncRNA）和信使 RNA（mRNA），并探讨其潜在的作用机制。

方法

利用高通量测序获得缺氧诱导和常氧 A549 肺癌细胞中 lncRNA 和 mRNA 的表达谱。随后，对差异表达分子进行生物信息学分析，包括功能富集分析、蛋白质-蛋白质相互作用（PPI）网络分析和竞争性内源性 RNA（ceRNA）分析。最后，采用实时定量 PCR（qPCR）验证关键 lncRNA 和 mRNA 的表达变化。

结果

在研究中，发现 1155 个 mRNAs 和 215 个 lncRNAs 在缺氧组和常氧组之间存在差异表达。功能富集分析表明，差异表达的 mRNAs 显著富集于多种通路，包括 p53 信号通路、DNA 复制和细胞周期。此外，还鉴定了关键的 lncRNA-miRNA-mRNA 关系，如 RP11-58O9.2-hsa-miR-6749-3p-XRCC2 和 SNAP25-AS1-hsa-miR-6749-3p-TENM4。值得注意的是，qPCR 检测结果表明，与常氧组相比，缺氧组中 SNAP25-AS1、RP11-58O9.2、TENM4 和 XRCC2 的表达下调，而 LINC01164、VLDLR-AS1、RP11-14I17.2 和 CDKN1A 的表达上调。

结论

本研究结果提示 SNAP25-AS1、RP11-58O9.2、TENM4、XRCC2、LINC01164、VLDLR-AS1、RP11-14I17.2 和 CDKN1A 可能参与了缺氧诱导的肺癌发生。这些关键的 lncRNA 和 mRNA 通过多种机制发挥作用，包括竞争性内源性 RNA（ceRNA）途径。