Determining the effect of chemotherapeutic treatment on changes in protein expression can provide important targets for overcoming resistance. Due to challenges in simultaneously measuring large numbers of proteins, a paucity of data exists on global changes. To overcome these challenges, we utilized microwestern arrays that allowed us to measure the abundance and modification state of hundreds of cell signaling and transcription factor proteins in cells following drug exposure. HapMap lymphoblastoid cell lines (LCLs) were exposed to cisplatin, a chemotherapeutic agent commonly used to treat testicular, head and neck, non-small cell lung, and gynecological cancers. We evaluated the expression of 259 proteins following 2, 6, and 12 hours of cisplatin treatment in two LCLs with discordant sensitivity to cisplatin. Of these 259 proteins, 66 displayed significantly different protein expression changes (p<0.05). Fifteen of these proteins were evaluated in a second pair of LCLs with discordant sensitivities to cisplatin; six demonstrated significant differences in expression. We then evaluated a subset of 63 proteins in a second set of LCLs with discordant sensitivity and 40% of those that were significant in the first pair were also significant in the second part with concordant directionality (p<0.05). We functionally validated one of the top proteins identified, PDK1, and demonstrated a synergistic relationship between cisplatin and a PDK1 inhibitor in multiple lung cancer lines. This study highlights the potential for identifying novel targets through an understanding of cellular changes in protein expression and modification following drug treatments.

中文翻译：

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顺铂治疗后新蛋白表达变化的鉴定及其在联合治疗中的应用

确定化学疗法对蛋白质表达变化的影响可以为克服耐药性提供重要的靶点。由于在同时测量大量蛋白质方面存在挑战，因此缺乏有关全球变化的数据。为了克服这些挑战，我们利用微western阵列，使我们能够测量药物暴露后细胞中数百种细胞信号和转录因子蛋白的丰度和修饰状态。将HapMap淋巴母细胞样细胞系（LCL）暴露于顺铂，顺铂是一种通常用于治疗睾丸癌，头颈癌，非小细胞肺癌和妇科癌症的化学治疗剂。我们评估了在两个对顺铂敏感性不一致的LCL中顺铂处理2、6和12小时后259种蛋白质的表达。在这259种蛋白质中，66个样品显示出显着不同的蛋白质表达变化（p <0.05）。在第二对对顺铂敏感性不一致的LCL中评估了其中的15种蛋白质。六位表现出明显的差异。然后，我们评估了第二组LCL中63种蛋白质的子集，其敏感性不一致，并且在第一对中显着的蛋白质中有40％在第二部分中也具有一致的方向性（p <0.05）。我们在功能上验证了鉴定出的顶级蛋白质之一PDK1，并证明了多种肺癌细胞系中顺铂和PDK1抑制剂之间的协同关系。这项研究突出了通过了解药物治疗后蛋白质表达和修饰的细胞变化来识别新靶标的潜力。