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.

中文翻译：

---

延长的无锚定寿命如何增强非小细胞肺癌细胞逃避失巢凋亡？– 与改变的细胞代谢组学的联系

恶性肿瘤细胞采用失巢凋亡抵抗来承受无锚定应激并引发癌症转移。目前尚不清楚不同时期的锚定丢失如何导致失巢凋亡抵抗、细胞迁移和癌细胞的代谢重编程。我们的研究表明，与野生型相比，将非小细胞肺癌 NCI-H460 细胞的无锚定寿命延长 7 天可增强失巢凋亡抵抗力，这表现在半衰期更长以及无需锚定的情况下生存和生长的能力细胞或那些失去锚定3天的细胞。虽然延长的无锚定寿命是导致存活细胞在 Transwell 测定的前 3 小时内进行快速 3 维迁移的攻击性增强的原因，但在 12 和 24 时检测到的 2 维表面迁移没有观察到显着影响。 h 通过伤口愈合方法。代谢组学分析显示，在所有 37 种已鉴定的物质中，六种物质（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着变化7 天无锚定期间的代谢。根据所有代谢物及其相关途径的显着性值、富集比和影响分数，三种主要代谢活动（非标准氨基酸代谢、细胞膜生物合成和氧化应激反应）提供了与失巢凋亡抗性的潜在联系。这些发现进一步加深了我们对肺癌细胞失巢凋亡抵抗性进化的认识，并确定了用于早期肺癌诊断的有希望的生物标志物。