Moreno-Sanchez Rafael, Vargas-Navarro Jorge Luis, Padilla-Flores Joaquin Alberto, Robledo-Cadena Diana Xochiquetzal, Granados-Rivas Juan Carlos, Taba Rutt, Terasmaa Anton, Auditano Giuseppe Leonardo, Kaambre Tuuli, Rodriguez-Enriquez Sara
Laboratorio de Control Metabólico, Carrera de Biología, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Tlalnepantla de Baz, Estado de México, México.
Laboratory of Chemical Biology, National Institute of Chemical Physics and Biophysics, Tallinn, Estonia.
Mini Rev Med Chem. 2025;25(4):319-339. doi: 10.2174/0113895575322436240924101642.
Analysis of the biochemical differences in the energy metabolism among bi-dimensional (2D) and tri-dimensional (3D) cultured cancer cell models and actual human tumors was undertaken. In 2D cancer cells, the oxidative phosphorylation (OxPhos) fluxes range is 2.5-19 nmol O2/min/mg cellular protein. Hypoxia drastically decreased OxPhos flux by 2-3 times in 2D models, similar to what occurs in mature multicellular tumor spheroids (MCTS), a representative 3D cancer cell model. However, mitochondrial protein contents and enzyme activities were significantly different between both models. Moreover, glycolytic fluxes were also significantly different between 2D and MCTS. The glycolytic flux range in 2D models is 1-34 nmol lactate/min/mg cellular protein, whereas in MCTS the range of glycolysis fluxes is 60-80 nmol lactate/min/mg cellular. In addition, sensitivity to anticancer canonical and metabolic drugs was greater in MCTS than in 2D. Actual solid human tumor samples show lower (1.6-4.5 times) OxPhos fluxes compared to normoxic 2D cancer cell cultures. These observations indicate that tridimensional organization provides a unique microenvironment affecting tumor physiology, which has not been so far faithfully reproduced by the 2D environment. Thus, the analysis of the resemblances and differences among cancer cell models undertaken in the present study raises caution on the interpretation of results derived from 2D cultured cancer cells when they are extended to clinical settings. It also raises awareness about detecting which biological and environmental factors are missing in 2D and 3D cancer cell models to be able to reproduce the actual human tumor behavior.
对二维（2D）和三维（3D）培养的癌细胞模型以及实际人类肿瘤之间能量代谢的生化差异进行了分析。在二维癌细胞中，氧化磷酸化（OxPhos）通量范围为2.5 - 19 nmol O2/分钟/毫克细胞蛋白。缺氧使二维模型中的OxPhos通量急剧下降2 - 3倍，这与成熟多细胞肿瘤球体（MCTS，一种代表性的三维癌细胞模型）中的情况类似。然而，两种模型之间的线粒体蛋白含量和酶活性存在显著差异。此外，二维模型和MCTS之间的糖酵解通量也存在显著差异。二维模型中的糖酵解通量范围为1 - 34 nmol乳酸/分钟/毫克细胞蛋白，而在MCTS中糖酵解通量范围为60 - 80 nmol乳酸/分钟/毫克细胞。此外，MCTS对抗癌经典药物和代谢药物的敏感性高于二维模型。与常氧二维癌细胞培养相比，实际人类实体肿瘤样本的OxPhos通量较低（低1.6 - 4.5倍）。这些观察结果表明，三维组织结构提供了一个影响肿瘤生理学的独特微环境，而二维环境迄今为止尚未如实地再现这一点。因此，本研究中对癌细胞模型之间异同的分析提醒人们，在将二维培养的癌细胞得出的结果推广到临床环境时要谨慎解释。这也提高了人们对检测二维和三维癌细胞模型中哪些生物和环境因素缺失以便能够再现实际人类肿瘤行为的认识。
