Wright Aaron T, Magnaldo Thierry, Sontag Ryan L, Anderson Lindsey N, Sadler Natalie C, Piehowski Paul D, Gache Yannick, Weber Thomas J
Omic Biological Applications, Pacific Northwest National Laboratory, Richland, Washington.
Mol Carcinog. 2015 Jun;54(6):473-84. doi: 10.1002/mc.22115. Epub 2013 Nov 27.
Human phenotypes that are highly susceptible to radiation carcinogenesis have been identified. Sensitive phenotypes often display robust regulation of molecular features that modify biological response, which can facilitate identification of the pathways/networks that contribute to pathophysiological outcomes. Here we interrogate primary dermal fibroblasts isolated from Gorlin syndrome patients (GDFs), who display a pronounced inducible tumorigenic response to radiation, in comparison to normal human dermal fibroblasts (NHDFs). Our approach exploits newly developed thiol reactive probes to define changes in protein thiol profiles in live cell studies, which minimizes artifacts associated with cell lysis. Redox probes revealed deficient expression of an apparent 55 kDa protein thiol in GDFs from independent Gorlin syndrome patients, compared with NHDFs. Proteomics tentatively identified this protein as aldehyde dehydrogenase 1A1 (ALDH1A1), a key enzyme regulating retinoic acid synthesis, and ALDH1A1 protein deficiency in GDFs was confirmed by Western blot. A number of additional protein thiol differences in GDFs were identified, including radiation responsive annexin family members and lamin A/C. Collectively, candidates identified in our study have plausible implications for radiation health effects and cancer susceptibility.
已确定对辐射致癌高度敏感的人类表型。敏感表型通常对修饰生物反应的分子特征具有强大的调控作用,这有助于识别导致病理生理结果的途径/网络。在这里,我们研究了从戈林综合征患者(GDFs)分离出的原代表皮成纤维细胞,与正常人皮肤成纤维细胞(NHDFs)相比,GDFs对辐射表现出明显的诱导致瘤反应。我们的方法利用新开发的硫醇反应探针来定义活细胞研究中蛋白质硫醇谱的变化,从而将与细胞裂解相关的假象降至最低。氧化还原探针显示,与NHDFs相比,来自独立戈林综合征患者的GDFs中一种明显的55 kDa蛋白质硫醇表达不足。蛋白质组学初步将该蛋白质鉴定为醛脱氢酶1A1(ALDH1A1),这是一种调节视黄酸合成的关键酶,Western印迹法证实了GDFs中ALDH1A1蛋白的缺乏。还发现了GDFs中许多其他蛋白质硫醇差异,包括辐射反应性膜联蛋白家族成员和核纤层蛋白A/C。我们研究中鉴定出的候选物对辐射健康影响和癌症易感性具有合理的意义。