Genovese Filippo, Gualandi Alessandra, Taddia Laura, Marverti Gaetano, Pirondi Silvia, Marraccini Chiara, Perco Paul, Pelà Michela, Guerrini Remo, Amoroso Maria Rosaria, Esposito Franca, Martello Andrea, Ponterini Glauco, D'Arca Domenico, Costi Maria Paola
C.I.G.S., University of Modena and Reggio Emilia , Via G. Campi 213/A, Modena 41125, Italy.
J Proteome Res. 2014 Nov 7;13(11):5250-61. doi: 10.1021/pr500510v. Epub 2014 Sep 29.
The preclinical study of the mechanism of action of anticancer small molecules is challenging due to the complexity of cancer biology and the fragmentary nature of available data. With the aim of identifying a protein subset characterizing the cellular activity of anticancer peptides, we used differential mass spectrometry to identify proteomic changes induced by two peptides, LR and [d-Gln(4)]LR, that inhibit cell growth and compared them with the changes induced by a known drug, pemetrexed, targeting the same enzyme, thymidylate synthase. The quantification of the proteome of an ovarian cancer cell model treated with LR yielded a differentially expressed protein data set with respect to untreated cells. This core set was expanded by bioinformatic data interpretation, the biologically relevant proteins were selected, and their differential expression was validated on three cis-platinum sensitive and resistant ovarian cancer cell lines. Via clustering of the protein network features, a broader view of the peptides' cellular activity was obtained. Differences from the mechanism of action of pemetrexed were inferred from different modulation of the selected proteins. The protein subset identification represents a method of general applicability to characterize the cellular activity of preclinical compounds and a tool for monitoring the cellular activity of novel drug candidates.
由于癌症生物学的复杂性以及现有数据的碎片化，抗癌小分子作用机制的临床前研究颇具挑战性。为了鉴定出能够表征抗癌肽细胞活性的蛋白质子集，我们使用差异质谱法来识别两种抑制细胞生长的肽LR和[d-Gln(4)]LR诱导的蛋白质组变化，并将其与一种已知药物培美曲塞（靶向同一种酶胸苷酸合成酶）诱导的变化进行比较。对用LR处理的卵巢癌细胞模型的蛋白质组进行定量分析，得到了相对于未处理细胞的差异表达蛋白质数据集。通过生物信息学数据解读扩展了这个核心数据集，选择了具有生物学相关性的蛋白质，并在三种顺铂敏感和耐药的卵巢癌细胞系上验证了它们的差异表达。通过对蛋白质网络特征进行聚类，我们对这些肽的细胞活性有了更全面的认识。从所选蛋白质的不同调节作用中推断出与培美曲塞作用机制的差异。蛋白质子集鉴定是一种具有普遍适用性的方法，可用于表征临床前化合物的细胞活性，也是监测新型候选药物细胞活性的工具。
