揭示 EIF5A 的作用：一种潜在的调节细胞周期的诊断标志物，与肺腺癌中的免疫浸润呈负相关。

Unlocking the role of EIF5A: A potential diagnostic marker regulating the cell cycle and showing negative correlation with immune infiltration in lung adenocarcinoma.

机构信息

College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, PR China; Key Laboratory of Zoonosis Prevention and Control of Guangdong Province, Guangzhou, Guangdong Province 510642, PR China; Key Laboratory of Zoonosis of Ministry of Agriculture and Rural Affairs, South China Agricultural University, Guangzhou, Guangdong Province 510642, PR China.

College of Agriculture, South China Agricultural University, Guangzhou, Guangdong Province 510642, PR China.

出版信息

Int Immunopharmacol. 2024 Jan 5;126:111227. doi: 10.1016/j.intimp.2023.111227. Epub 2023 Nov 15.

DOI:10.1016/j.intimp.2023.111227

PMID:37977067

Abstract

BACKGROUND

Despite EIF5A upregulation related to tumor progression in LUAD (lung adenocarcinoma), the underlying mechanisms remain elusive. In addition, there are few comprehensive analyses of EIF5A in LUAD.

METHODS

We investigated the EIF5A expression level in LUAD patients using data from the TCGA and GEO databases. We employed qRT-PCR and western blot to verify EIF5A expression in cell lines, while immunohistochemistry was utilized for clinical sample analysis. We analyzed EIF5A expression in tumor-infiltrating immune cells using the TISCH database and assessed its association with immune infiltration in LUAD using the "ESTIMATE" R package. Bioinformatics approaches were developed to discover the EIF5A-related genes and explore EIF5A potential mechanisms in LUAD. Proliferation ability was verified through CCK-8, clone formation, and EdU assays, while flow cytometry assessed apoptosis and cell cycle. Western blot was used to detect the expression of pathway-related proteins.

RESULTS

EIF5A was significantly upregulated in LUAD. Moreover, we constructed a MAZ-hsa-miR-424-3p-EIF5A transcriptional network. We explored the potential mechanism of EIF5A in LUAD and further investigated the cAMP signaling pathway and the cell cycle. Finally, we proved that EIF5A silencing induced G1/S Cell Cycle arrest, promoted apoptosis, and inhibited proliferation via the cAMP/PKA/CREB signaling pathway.

CONCLUSION

EIF5A serves as a prognostic biomarker with a negative correlation to immune infiltrates in LUAD. It regulated the cell cycle in LUAD by inhibiting the cAMP/PKA/CREB signaling pathway.

摘要

背景

尽管 EIF5A 在 LUAD（肺腺癌）中的上调与肿瘤进展有关，但潜在机制仍不清楚。此外，对 LUAD 中 EIF5A 的综合分析很少。

方法

我们使用 TCGA 和 GEO 数据库中的数据研究了 LUAD 患者的 EIF5A 表达水平。我们使用 qRT-PCR 和 Western blot 验证了细胞系中的 EIF5A 表达，同时使用免疫组织化学分析了临床样本。我们使用 TISCH 数据库分析了肿瘤浸润免疫细胞中的 EIF5A 表达，并使用“ESTIMATE”R 包评估了其与 LUAD 中免疫浸润的相关性。我们开发了生物信息学方法来发现 EIF5A 相关基因，并探索 EIF5A 在 LUAD 中的潜在机制。通过 CCK-8、克隆形成和 EdU 测定验证增殖能力，通过流式细胞术评估细胞凋亡和细胞周期。Western blot 用于检测通路相关蛋白的表达。

结果

EIF5A 在 LUAD 中显著上调。此外，我们构建了 MAZ-hsa-miR-424-3p-EIF5A 转录网络。我们探讨了 EIF5A 在 LUAD 中的潜在机制，并进一步研究了 cAMP 信号通路和细胞周期。最后，我们证明 EIF5A 沉默通过 cAMP/PKA/CREB 信号通路诱导 G1/S 细胞周期停滞、促进细胞凋亡和抑制增殖。