Podsednik Allison, Xu He N, Li Lin Z
Britton Chance Laboratory of Redox Imaging, Department of Radiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Transl Breast Cancer Res. 2024 Oct 31;5:27. doi: 10.21037/tbcr-24-36. eCollection 2024.
The redox status of nicotinamide adenine dinucleotide (NAD; including oxidized form NAD and reduced form NADH) plays key roles in both health and disease and has been actively studied to develop cancer biomarkers and therapeutic strategies. With the optical redox imaging (ORI) technique, we have been investigating the relationship of NADH redox status, reactive oxygen species (ROS), and invasiveness in breast cancer cell cultures, and have associated higher invasiveness with more oxidized NADH redox state. However, the cell cultures may have phenotypic drift and metabolic change with increased passage numbers.
We investigated the passage-dependence of NADH redox status and ROS levels in two triple-negative breast cancer (TNBC) cell cultures: the more invasive/metastatic MDA-MB-231 and the less invasive/metastatic HCC1806 cell lines. We measured the NADH redox status, redox plasticity, and cytoplasmic and mitochondrial ROS levels under the basal condition and metabolic perturbations of the mitochondrial electron transport chain. We evaluated the dependence of redox and ROS profiles on the cell passage number by comparing the early (<20 passages) with the late (>60 passages) passage cells.
(I) NADH redox and ROS baselines are stable and independent of cell passage number, but can vary with passage number under metabolic perturbations depending on specific perturbation and cell line; (II) NADH redox status and intracellular ROS levels can change discordantly in cancer cells; (III) under both basal and metabolically perturbed conditions, the more invasive cell line has a more oxidized NADH redox status with a higher basal cytoplasmic ROS level than the less invasive line, regardless of passage number.
The general correlation between redox, ROS, and invasiveness in studied TNBC cells is not very sensitive to passage number. These results indicate that NADH redox and basal ROS status in TNBC likely reflect the intrinsic progressive nature of TNBC cells.
烟酰胺腺嘌呤二核苷酸（NAD，包括氧化型NAD和还原型NADH）的氧化还原状态在健康和疾病中均起着关键作用，并且人们一直在积极研究其用于开发癌症生物标志物和治疗策略。通过光学氧化还原成像（ORI）技术，我们一直在研究乳腺癌细胞培养物中NADH氧化还原状态、活性氧（ROS）与侵袭性之间的关系，并发现侵袭性越高，NADH氧化还原状态越氧化。然而，随着传代次数的增加，细胞培养物可能会出现表型漂移和代谢变化。
我们研究了两种三阴性乳腺癌（TNBC）细胞培养物中NADH氧化还原状态和ROS水平对传代的依赖性：侵袭性/转移性更强的MDA-MB-231细胞系和侵袭性/转移性较弱的HCC1806细胞系。我们在基础条件以及线粒体电子传递链的代谢扰动下测量了NADH氧化还原状态、氧化还原可塑性以及细胞质和线粒体ROS水平。通过比较早期（<20代）和晚期（>60代）传代细胞，我们评估了氧化还原和ROS谱对细胞传代次数的依赖性。
（I）NADH氧化还原和ROS基线是稳定的，且与细胞传代次数无关，但在代谢扰动下，根据特定的扰动和细胞系，它们可能会随传代次数而变化；（II）癌细胞中NADH氧化还原状态和细胞内ROS水平可能会不一致地变化；（III）在基础条件和代谢扰动条件下，无论传代次数如何，侵袭性更强的细胞系比侵袭性较弱的细胞系具有更氧化的NADH氧化还原状态以及更高的基础细胞质ROS水平。
在研究的TNBC细胞中，氧化还原、ROS与侵袭性之间的一般相关性对传代次数不是很敏感。这些结果表明，TNBC中的NADH氧化还原和基础ROS状态可能反映了TNBC细胞的内在进展特性。
