【作者】 汪洋；

【导师】 朱亮；

【作者基本信息】 上海交通大学 ， 药学， 2019， 硕士

【摘要】 肺癌是世界上发生率和死亡率均居前列的恶性肿瘤。目前,分子靶向药物EGFR-TKI已进入临床治疗肺癌并取得显著疗效,但是一段时间后几乎不可避免出现耐药现象,引起肺癌复发和不良预后。肿瘤耐药过程中,代谢重编程起到重要作用。活性醛作为一类可以对细胞产生氧化损伤的代谢物,它与活性氧相互促进、相互影响,形成“活性醛-活性氧”轴。大量研究表明肿瘤耐药与醛脱氢酶ALDH及活性氧相关,但是针对耐药细胞活性醛的研究较少,由于现有检测方法选择性且灵敏度不够,肺癌耐药细胞中活性醛含量的变化也是不明确的。代谢组学作为一门新兴的组学,已成为研究肿瘤耐药机制的新方向。本研究从代谢组学理论出发,基于UPLC-QQQ/MS技术平台,通过衍生化建立了靶向细胞内活性醛的新分析方法,提高了检测灵敏度,扩大了检测活性醛种类,发现耐药细胞中整体活性醛含量下降。为了研究耐药细胞重编程代谢通路与活性醛的关系,我们基于UPLC-QTOF/MS技术平台分析了耐药细胞的代谢改变,其中GSH代谢变化最为显著。因此,我们针对GSH代谢通路,开发出新的靶向分析方法,发现耐药细胞可以增强GSH代谢通路并且能耐受胞内较高的氧化状态。本研究开发的代谢组学检测方法具有普适性,也为肺癌耐药机制提供了新的代谢组学研究解释。更多还原

【Abstract】 Lung cancer is the leading cause of cancer-related death in the world today.EGFR Tyrosine kinase inhibitors(EGFR-TKI)has been proposed for the treatment of lung adenocarcinoma targeting EGFR mutations.Unfortunately,drug resistance is inevitable in lung cancer patients,leading to tumor recurrence and poor prognosis after using the targeted drug for a period of time.Metabolic reprogramming plays a vital role in tumor resistance.As a kind of metabolite which can cause oxidative damage to cells,reactive aldehyde promotes and interacts with active oxygen to form "reactive aldehyde-reactive oxygen" axis.Many studies have suggested that tumor resistance is associated with aldehyde dehydrogenase(ALDH)and reactive oxygen species,but there are few studies on reactive aldehydes in drug-resistant cells.Due to the limitations of existing detection methods and insufficient sensitivity,change in the level of reactive aldehydes in lung cancer resistant cells is unclear.Metabonomics,as a new histology,has become a new direction to study the mechanism of drug resistance in tumors.Based on the ultra performance liquid chromatography tandem triple-quadrupole mass spectrometry(UPLC-QQQ/MS)technology platform,we have established an analytical method for detecting the level of reactive aldehydes in cells by derivatization,which improves the detection sensitivity and expands the number of active aldehydes detected.This study suggests a decrease in the level of reactive aldehydes in drug-resistant cells.To investigate the relationship between altered metabolic pathways and reactive aldehydes in drug-resistant cells,we analyzed the metabolic profiling in drug-resistant cells by non-targeted metabolomics based on Ultraperformance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS).We found that glutathione metabolism is most prominently altered in resistant cells.Therefore,we have developed a fast-processing,high-throughput analytical method for target metabolites in GSH metabolic pathway.It is revealed that drug-resistant cells can enhance the GSH metabolic pathway and tolerate a lower intracellular reduction state.This study developed a new targeted analytical method for reactive aldehydes and GSH metabolic pathways,as well as nontargeted analytical method for drug-resistant cell models,which can be applied to other cell models to support research.It also provides a new metabolomics study explanation for lung cancer resistance mechanisms.更多还原