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

描述(申请人提供)：建议使用LC-MS/MS(LC-MS/MS)来表征肿瘤细胞对化疗的反应。现代LC-MS/MS技术采用电喷雾界面和三重四极杆检测，提供了前所未有的灵敏度、选择性和测量多种产品的能力。建议将LC MS/MS技术应用于抗癌药物引起的DNA修饰，包括直接和/或间接损伤、各种类型的链断裂和细胞凋亡产物的检测。使用一种在消除背景方面非常有效的样品制备方法，在单个样品中同时测量这种不同的DNA损伤和修饰是可行的。以顺铂暴露细胞的LC-MS/MS测量为例，验证了该方法的可行性。建议LC-MS/MS法可应用于手术中获得的细胞，采用目前用于检测耐药性的培养方法。LC-MS/MS分析揭示的细胞功能差异将得到基因表达阵列研究的补充。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.