Dahl Christina, Guldberg Per, Abildgaard Cecilie
Danish Cancer Society Research Center, Copenhagen, Denmark.
Methods Mol Biol. 2019;2019:171-180. doi: 10.1007/978-1-4939-9585-1_12.
Dysregulation of retinoic acid signaling is implicated in several human cancer types, including melanoma where the gene encoding retinoic acid receptor beta (RARβ) is frequently silenced by promoter hypermethylation. In this chapter, we describe some of the experimental procedures that we have used to characterize the role of RARβ signaling on the regulation of cellular metabolism in melanoma. Central to these studies is the use of the Seahorse XF Analyzer, which allows real-time assessment of the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in cultured cells as readouts for oxidative phosphorylation and glycolysis, respectively. The levels of RARβ signaling can be modulated using RARβ agonists (e.g., all-trans retinoic acid) and antagonists (e.g., LE135). The bioenergetic profiles of melanoma cells in response to RARβ modulators and other metabolic modifiers can be the basis for defining new therapeutic strategies.
维甲酸信号失调与多种人类癌症类型有关，包括黑色素瘤，其中编码维甲酸受体β（RARβ）的基因经常因启动子高甲基化而沉默。在本章中，我们描述了一些实验程序，这些程序用于表征RARβ信号在黑色素瘤细胞代谢调节中的作用。这些研究的核心是使用Seahorse XF分析仪，该分析仪可以实时评估培养细胞中的氧消耗率（OCR）和细胞外酸化率（ECAR），分别作为氧化磷酸化和糖酵解的读数。RARβ信号水平可以使用RARβ激动剂（如全反式维甲酸）和拮抗剂（如LE135）进行调节。黑色素瘤细胞对RARβ调节剂和其他代谢修饰剂的生物能量谱可以作为定义新治疗策略的基础。
