D'Aguanno Simona, Barcaroli Daniela, Rossi Claudia, Zucchelli Mirco, Ciavardelli Domenico, Cortese Claudio, De Cola Antonella, Volpe Silvia, D'Agostino Daniela, Todaro Matilde, Stassi Giorgio, Di Ilio Carmine, Urbani Andrea, De Laurenzi Vincenzo
Department of Experimental and Clinical Sciences, "G. d'Annunzio University" , Via dei Vestini 31, Chieti-Pescara 66100, Italy.
J Proteome Res. 2014 Apr 4;13(4):2120-36. doi: 10.1021/pr4012574. Epub 2014 Mar 19.
p63 is an important regulator of epithelial development expressed in different variants containing (TA) or lacking (ΔN) the N-terminal transactivation domain. The different isoforms regulate stem-cell renewal and differentiation as well as cell senescence. Several studies indicate that p63 isoforms also play a role in cancer development; however, very little is known about the role played by p63 in regulating the cancer stem phenotype. Here we investigate the cellular signals regulated by TAp63 and ΔNp63 in a model of epithelial cancer stem cells. To this end, we used colon cancer stem cells, overexpressing either TAp63 or ΔNp63 isoforms, to carry out a proteomic study by chemical-labeling approach coupled to network analysis. Our results indicate that p63 is implicated in a wide range of biological processes, including metabolism. This was further investigated by a targeted strategy at both protein and metabolite levels. The overall data show that TAp63 overexpressing cells are more glycolytic-active than ΔNp63 cells, indicating that the two isoforms may regulate the key steps of glycolysis in an opposite manner. The mass-spectrometry proteomics data of the study have been deposited to the ProteomeXchange Consortium ( http://proteomecentral.proteomexchange.org ) via the PRIDE partner repository with data set identifiers PXD000769 and PXD000768.
p63是上皮发育的重要调节因子，以包含（TA）或缺乏（ΔN）N端反式激活结构域的不同变体形式表达。不同的异构体调节干细胞的更新与分化以及细胞衰老。多项研究表明，p63异构体在癌症发展中也发挥作用；然而，关于p63在调节癌症干细胞表型中所起的作用却知之甚少。在此，我们在一个上皮癌干细胞模型中研究由TAp63和ΔNp63调节的细胞信号。为此，我们使用过表达TAp63或ΔNp63异构体的结肠癌干细胞，通过与网络分析相结合的化学标记方法进行蛋白质组学研究。我们的结果表明，p63参与了包括代谢在内的广泛生物学过程。我们在蛋白质和代谢物水平上通过靶向策略对此进行了进一步研究。总体数据显示，过表达TAp63的细胞比过表达ΔNp63的细胞具有更高的糖酵解活性，这表明这两种异构体可能以相反的方式调节糖酵解的关键步骤。该研究的质谱蛋白质组学数据已通过PRIDE合作伙伴储存库存入蛋白质组交换联盟（http://proteomecentral.proteomexchange.org），数据集标识符为PXD000769和PXD000768。
