Institute of Bioinformatics and Structural Biology and Department of Medical Sciences, National Tsing Hua University, Hsinchu 30013, Taiwan.
Toxicol Sci. 2014 Jun;139(2):396-406. doi: 10.1093/toxsci/kfu051. Epub 2014 Mar 27.
The nucleus is a key organelle in mammary cells, which is responsible for several cellular functions including cell proliferation, gene expression, and cell survival. In addition, the nucleus is the primary targets of doxorubicin treatment. In the current study, low-abundance nuclear proteins were enriched for proteomic analysis by using a state-of-the-art two-dimensional differential gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) strategy to compare and identify the nuclear protein profiling changes responsible for the development of doxorubicin resistance in human uterine cancer cells. The results of the nuclear proteomic analysis indicated that more than 2100 protein features were resolved from an equal pooled amount of three purified nuclear proteins and 117 differentially expressed spots were identified. Of these 117 identified proteins, 48 belonged to nuclear proteins and a positive correlation was observed between the expression levels of 32 of these nuclear proteins and an increase in drug resistance. According to our review of relevant research, nuclear proteins such as DNA repair protein XRCC3 (XRCC3) have not been reported to play roles in the formation of doxorubicin resistance. Previous studies have used RNA interference and cell viability analysis to evidence the essential roles of XRCC3 on its potency in the formation of doxorubicin resistance. To sum up, our nuclear proteomic approaches enabled us to identify numerous proteins, including XRCC3, involved in various drug-resistance-forming mechanisms. Our results provide potential diagnostic markers and therapeutic candidates for treating doxorubicin-resistant uterine cancer.
核是乳腺细胞的一个关键细胞器，负责包括细胞增殖、基因表达和细胞存活在内的多种细胞功能。此外，核是多柔比星治疗的主要靶点。在本研究中，通过使用最先进的二维差异凝胶电泳（2D-DIGE）和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）策略，富集低丰度核蛋白进行蛋白质组学分析，以比较和鉴定导致人子宫癌细胞多柔比星耐药性发展的核蛋白谱变化。核蛋白质组分析的结果表明，从等量的三种纯化核蛋白中可以分辨出 2100 多个蛋白质特征，鉴定出 117 个差异表达斑点。在这 117 种鉴定的蛋白质中，有 48 种属于核蛋白，其中 32 种核蛋白的表达水平与药物耐药性的增加呈正相关。根据我们对相关研究的回顾，核蛋白如 DNA 修复蛋白 XRCC3（XRCC3）尚未被报道在多柔比星耐药性的形成中发挥作用。以前的研究已经使用 RNA 干扰和细胞活力分析来证明 XRCC3 在形成多柔比星耐药性方面的效力的重要作用。总之，我们的核蛋白质组学方法使我们能够鉴定出许多参与各种耐药形成机制的蛋白质，包括 XRCC3。我们的结果为治疗多柔比星耐药性子宫癌提供了潜在的诊断标志物和治疗候选物。
