LC-MS/MS Assessments of Apoptosis/DNA Damage/Repair

描述（由申请方提供）：拟采用液相色谱-串联质谱法（LC-MS/MS）表征肿瘤细胞对化疗的反应。现代LC-MS/MS技术采用电喷雾接口和三重四极杆检测，提供了前所未有的灵敏度，选择性和测量多种产品的能力。建议LCMS/ MS技术适用于测量抗癌药物诱导的DNA修饰，包括直接和/或间接损伤，各种类型的链断裂和凋亡产物。使用在消除背景方面非常有效的样品制备方法，可以在单个样品中同时测量这种多样的DNA损伤和修饰。该方法的可行性在应用中使用暴露于顺铂的细胞上的LC-MS/MS测量作为示例来证明。建议LC-MS/MS分析可应用于使用目前用于测量耐药性的培养方法在手术中获得的细胞。通过LC-MS/MS分析揭示的细胞功能差异将通过基因表达的阵列研究进行补充。第一个临床应用将是卵巢癌组织。

DESCRIPTION (provided by applicant): It is proposed that Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) be utilized to characterize the response of tumor cells to chemotherapy. Modern LC-MS/MS technology, employing an electrospray interface and triple quadrapole detection, provides unprecedented sensitivity, selectivity and the capability for measuring multiple products. It is proposed that LCMS/ MS technology be adapted to the measurement of DNA modifications induced by anticancer drugs including direct and/or indirect damage, various types of strand breaks and the products of apoptosis. Using a method of sample preparation that is highly effective in eliminating background it is feasible to measure simultaneously this variety of DNA lesions and modifications in a single sample. The feasibility of this approach is demonstrated in the application using LC-MS/MS measurements on cells exposed to cisplatin as an example. It is proposed that LC-MS/MS assay can be applied to cells obtained at surgery suing the culture methods presently employed to measure drug resisitivity. Differences in cell function revealed by LC-MS/MS assay will be complemented by array studies of gene expression. The first clinical application will be ovarian cancer tissue.