摘 要：GCN2-eIF2α通路可以调节细胞生长，是综合应激反应（ISR）的关键组成部分。在恶性肿瘤中，肿瘤细胞处于慢性缺氧和营养缺乏的环境中，此时氨基酸分解代谢水平升高，GCN2-eIF2α通路被激活，蛋白质合成被抑制以维持自身状态。本研究以9例肺癌患者非小细胞肺癌（NSCLC）的癌组织和与之匹配的正常组织样本为研究对象，采用组织学、转录组学、qRT-PCR和蛋白质免疫印迹技术联合分析，对GCN2-eIF2α通路调控癌细胞增殖的分子机制进行研究。石蜡切片结果表明，肺癌患者的癌组织肺泡细胞生长密集，肺泡间隔窄，细胞核染色不均，具有典型的恶性组织病理学特征。转录组研究表明，转录组中基因本体论（GO）和京都基因与基因组百科全书（KEGG）的富集结果确定了样本间的富集通路和差异基因，确定了GCN2-eIF2α通路在肺癌发生中的重要作用。进一步从中筛选出关键基因进行分子水平和蛋白水平上的验证，qRT-PCR结果显示，其在癌组织细胞和正常组织细胞中均有表达，且表达量在癌组织细胞中大多呈上调趋势（P<0.05），与转录组中的基因丰度分析结果一致。在蛋白水平的检测中，一般性调控阻遏蛋白激酶2（GCN2）和真核翻译起始因子2α激酶（eIF2α）的表达量在癌组织细胞中明显高于正常组织。这表明，GCN2-eIF2α通路与肺癌的发生密切相关。本研究可为eIF2α、GCN2作为新的肿瘤治疗靶点提供参考依据，也提供了一种通过抑制GCN2磷酸化eIF2α，从而抑制癌细胞发展的可能性，为解决当前抗肿瘤疗法的局限性提供了一种答案。
关键词：肺癌；GCN2-eIF2α通路；转录组学；癌症发生机制；靶向肿瘤治疗
中图分类号：R734.2 文献标志码：A DOI：10.3969/j.issn.1007-7146.2025.06.007
Abstract: The GCN2-eIF2α pathway regulates cell growth and serves as a key component of the integrated stress response (ISR). In malignant tumors, cancer cells reside in a microenvironment characterized by chronic hypoxia and nutrient deprivation. Under such conditions, the level of amino acid catabolism increases, which activates the GCN2-eIF2α pathway and inhibits protein synthesis to maintain the cellular homeostasis of cancer cells. In this study, a total of 9 pairs of tissue samples, including cancerous tissues and their matched adjacent normal tissues, were collected from patients diagnosed with non-small cell lung cancer (NSCLC). A combined analysis using histology, transcriptomics, qRT-PCR, and Western blotting was performed to investigate the molecular mechanism by which the GCN2-eIF2α pathway regulates cancer cell proliferation. Results of paraffin sections showed that in the tumor tissues of lung cancer patients, alveolar cells were densely packed, alveolar septa were narrow, and nuclear staining was heterogeneous, exhibiting typical malignant histopathological features. Transcriptomic studies revealed that the enriched pathways and differentially expressed genes among samples were identified based on gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment results from the transcriptome, which confirmed that the GCN2-eIF2α pathway plays an important role in lung carcinogenesis. Further screening was performed to select key genes for validation at the molecular and protein levels. qRT-PCR results demonstrated that these genes were expressed in both cancerous and normal tissue cells, with their expression levels mostly upregulated in cancerous tissue cells (P<0.05), which was consistent with the gene abundance analysis results in the transcriptome. At the protein level, the expression levels of general control nonderepressible 2 kinase (GCN2) and eukaryotic translation initiation 2α (eIF2α) in cancerous tissues were significantly higher than those in adjacent normal tissues, indicating that the GCN2-eIF2α pathway is implicated in lung carcinogenesis. This study can provide a reference for eIF2α and GCN2 to serve as new therapeutic targets for tumors, and also propose the possibility of inhibiting cancer cell progression by suppressing GCN2-mediated phosphorylation of eIF2α, thereby offering a solution to address the limitations of current anti-tumor therapies.
Key words: lung cancer; GCN2-eIF2α pathway; transcriptomics; cancer pathogenesis; targeted tumor therapy
（Acta Laser Biology Sinica, 2025, 34(6): 530-540）
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