利用细胞外流量分析和物质平衡对Rab25在能量代谢和癌症中的作用进行表征。
Characterization of the role Rab25 in energy metabolism and cancer using extracellular flux analysis and material balance.
作者信息
Mitra Shreya, Molina Jennifer, Mills Gordon B, Dennison Jennifer B
机构信息
Department of Systems Biology, UT MD Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX, 77054, USA,
出版信息
Methods Mol Biol. 2015;1298:195-205. doi: 10.1007/978-1-4939-2569-8_17.
Rab25, by altering trafficking of critical cellular resources, influences cell metabolism and survival during stress conditions. Overall, perturbations in the vesicular trafficking machinery change cellular bioenergetics that can be directly measured in real time as Oxygen Consumption Rate, OCR (mitochondrial respiration) and Extracellular Acidification Rate, ECAR (glycolysis) by an extracellular flux analyzer (XF96, Seahorse Biosciences, MA). Additionally, overall turnover of glucose, lactate, as well as glutamine and glutamate can be measured biochemically using the YSI2900 Biochemistry Analyzer (YSI Incorporated, Life Sciences, OH). A combination of these two methods allows a precise and quantitative approach to interrogate the role of Rab25 as well as other Rab GTPases in central carbon energy metabolism.
Rab25 通过改变关键细胞资源的运输，在应激条件下影响细胞代谢和存活。总体而言，囊泡运输机制的扰动会改变细胞生物能量学，这可以通过细胞外通量分析仪（XF96，海马生物科学公司，马萨诸塞州）实时直接测量为氧消耗率（OCR，线粒体呼吸）和细胞外酸化率（ECAR，糖酵解）。此外，可以使用 YSI2900 生物化学分析仪（YSI 公司，生命科学部，俄亥俄州）通过生化方法测量葡萄糖、乳酸以及谷氨酰胺和谷氨酸的总体周转率。这两种方法的结合提供了一种精确且定量的方法来探究 Rab25 以及其他 Rab GTP 酶在中心碳能量代谢中的作用。
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