Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai, 201203, China; University of Chinese Academy of Sciences, Number 19A Yuquan Road, Beijing, 100049, China.
Department of Analytical Chemistry and CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai, 201203, China.
Biochem Biophys Res Commun. 2018 Nov 10;505(4):1112-1120. doi: 10.1016/j.bbrc.2018.10.065. Epub 2018 Oct 15.
Dermal fibroblast is one of the major constitutive cells of skin and plays a central role in skin senescence. The replicative senescence of fibroblasts may cause skin aging, bad wound healing, skin diseases and even cancer. In this study, a label-free quantitative proteomic approach was employed to analyzing the serial passaged human skin fibroblast (CCD-1079Sk) cells, resulting in 3371 proteins identified. Of which, 280 proteins were significantly changed in early passage (6 passages, P6), middle passage (12 passages, P12) and late passage (21 passages, P21), with a time-dependent decrease or increase tendency. Bioinformatic analysis demonstrated that the chromosome condensin complex, including structural maintenance of chromosomes protein 2 (SMC2) and structural maintenance of chromosomes protein 4 (SMC4), were down-regulated in the serially passaged fibroblast cells. The qRT-PCR and Western Blot experiments confirmed that the expression of these two proteins were significantly down-regulated in a time-dependent manner in the subculture of human skin fibroblasts (HSFb cells). In summary, we used serially passaged human skin fibroblast cells coupled with quantitative proteomic approach to profile the protein expression pattern in the temporal progress of replicative senescence in HSFb cells and revealed that the down-regulation of the chromosome condensin complex subunits, such as SMC2 and SMC4, may play an important role in the fibroblast senescence.
真皮成纤维细胞是皮肤的主要固有细胞之一，在皮肤衰老中起着核心作用。成纤维细胞的复制性衰老可能导致皮肤老化、伤口愈合不良、皮肤疾病甚至癌症。在这项研究中，我们采用了一种无标记的定量蛋白质组学方法来分析连续传代的人皮肤成纤维细胞（CCD-1079Sk），结果鉴定出 3371 种蛋白质。其中，280 种蛋白质在早期传代（6 代，P6）、中期传代（12 代，P12）和晚期传代（21 代，P21）时发生了显著变化，呈时间依赖性的减少或增加趋势。生物信息学分析表明，染色体凝聚复合物，包括结构维持染色体蛋白 2（SMC2）和结构维持染色体蛋白 4（SMC4），在连续传代的成纤维细胞中下调。qRT-PCR 和 Western Blot 实验证实，这两种蛋白质的表达在人皮肤成纤维细胞（HSFb 细胞）的亚培养中呈时间依赖性显著下调。综上所述，我们使用连续传代的人皮肤成纤维细胞结合定量蛋白质组学方法，描绘了 HSFb 细胞复制性衰老过程中蛋白质表达模式的时间进程，并揭示了染色体凝聚复合物亚基如 SMC2 和 SMC4 的下调可能在成纤维细胞衰老中起重要作用。
