恶性细胞采用缺陷粘液抵抗以保持不受锚定应激的幸存和癌症转移。关于锚定损失的不同时间段如何影响缺陷性抵抗、细胞迁移和癌细胞代谢重编程，至今尚不清楚。我们的研究表明，将非小细胞肺癌NCI-H460细胞的无锚定寿命延长至7天，增强了缺陷性抵抗，表现为更长的半衰期和在无锚定条件下生存和生长的能力，与野生型细胞或失去锚定3天的细胞相比。虽然延长的无锚定寿命使存活细胞具有更强的侵略性特征，在转孔法实验的前3个小时内能够快速进行三维迁移，但通过伤口愈合方法检测到的2维表面迁移在12和24小时内未观察到明显影响。代谢组学分析显示，在7天的无锚定期间，37个已鉴定的代谢物中有六个（草酸、胆固醇、1-乙基吡咯烷、1-(3-甲基丁基)-2,3,4,6-四甲基苯、β-丙氨酸和腐胺）的细胞内水平发生显著变化。根据所有代谢物及其相关通路的显著性值、富集比率和影响分数，三种主要代谢活性（非标准氨基酸代谢、细胞膜生物合成和氧化应激响应）与缺陷性抵抗存在潜在联系。这些发现进一步增加了对肺癌细胞缺陷性抵抗演化的认识，并确定了早期肺癌诊断的有希望的生物标志物。 © 2023. Sociedad de Biologia de Chile.

Malignant cells adopt anoikis resistance to survive anchorage-free stresses and initiate cancer metastasis. It is still unknown how varying periods of anchorage loss contribute to anoikis resistance, cell migration, and metabolic reprogramming of cancerous cells.Our study demonstrated that prolonging the anchorage-free lifetime of non-small-cell lung cancer NCI-H460 cells for 7 days strengthened anoikis resistance, as shown by higher half-life and capability to survive and grow without anchorage, compared to wild-type cells or those losing anchorage for 3 days. While the prolonged anchorage-free lifetime was responsible for the increased aggressive feature of survival cells to perform rapid 3-dimensional migration during the first 3 h of a transwell assay, no significant influence was observed with 2-dimensional surface migration detected at 12 and 24 h by a wound-healing method. Metabolomics analysis revealed significant alteration in the intracellular levels of six (oxalic acid, cholesterol, 1-ethylpyrrolidine, 1-(3-methylbutyl)-2,3,4,6-tetramethylbenzene, β-alanine, and putrescine) among all 37 identified metabolites during 7 days without anchorage. Based on significance values, enrichment ratios, and impact scores of all metabolites and their associated pathways, three principal metabolic activities (non-standard amino acid metabolism, cell membrane biosynthesis, and oxidative stress response) offered potential links with anoikis resistance.These findings further our insights into the evolution of anoikis resistance in lung cancer cells and identify promising biomarkers for early lung cancer diagnosis.© 2023. Sociedad de Biologia de Chile.