The study of drug resistance mechanisms of breast cancer cells by quantitative mass spectrometry
作者单位：State Key Laboratory of ProteomicsBeijing Proteome Research CenterNational Center for Protein Sciences (Beijing)Beijing Institute of Lifeomics Lunenfeld Tanenbaum Research InstituteMount Sinai Hospital McGill University Health Centre Department of Molecular GeneticsUniversity of Toronto
会议名称：《2018年中国质谱学术大会（CMSC2018）》
会议日期：2018年
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
关 键 词：Quantitative mass spectrometry Drug resistance Dynamic Signaling Breast cancer cell
摘 要：The SHC1 adaptor protein directly binds to all members of the HER/ERBB receptor tyrosine kinase family and forms a critical link between the receptors and downstream effectors1,2. Since SHC1 only binds to receptors in their activated states, it is well positioned to monitor the translation of signals from the receptors to downstream targets which is of critical importance in ERBB-driven cancers. We affinity purified SHC1 followed by parallel reaction monitoring(PRM)-based quantification to measure changes in SHC1 complex formation in 19 cancer cell lines with various ERBB levels1. We targeted 35 SHC interactors for quantitation. Our data showed that the cell lines varied significantly in the type and number of proteins(from 6 to 32 proteins) that were recruited to SHC1. We further captured temporal profiles of the SHC1 complexes before and during EGF stimulation in four ERBB2/HER2 positive breast cancer cell lines(SKBR3, BT-474, HCC-1954, HCC-1419) with different sensitivity towards trastzumab3, a clinical inhibitor for ERBB2/HER2. We detected significantly stronger association of promitogenic components of SHC1 complexes at basal level in drug-sensitive cells over drug-resistant cells. This pattern is not observed for components involved in negatively regulation ERBB signaling. Moreover, the duration, but not the maximum level, of the association for pro-mitogenic components is apparently longer in drug-sensitive cells, suggesting defects in signal down regulation in these cells. Not all of these observations could be predicted from either m RNA or protein expression levels. Taking together, our analysis of dynamic SHC1 complex formation supports SHC1 as a function-base probe for monitoring ERBB activation status and a potential diagnostic tool for drug sensitivity prediction.