长期细胞培养是一种重要的生物学方法,但其特征还在于细胞形态的退化。增殖率,和功能。要系统地探讨这一现象,我们对AC16和HUVECs的两种心血管细胞系进行了综合蛋白质组学-代谢组学测量.第18段培养,即,G18,通过细胞代谢谱显示为转折点,其中代谢组学的变化证明了能量的功能障碍,氨基酸,和核糖核苷酸代谢代谢途径。尽管活跃的蛋白质网络显示线粒体丰富的AC16和氧化/硝化敏感的HUVEC表明不同的变性模式,G18和G30蛋白质组学通过信号转导过程证明衰老,白细胞介素信号,程序性细胞死亡,细胞对刺激的反应,细胞周期,mRNA剪接,和翻译。一些关键蛋白(RPS8,HNRNPR,SOD2,LMNB1,PSMA1,DECR1,GOT2,OGDH,PNP,CBS,ATIC,和IMPDH2)和代谢物(L-谷氨酸,鸟嘌呤,柠檬酸,鸟苷,鸟苷二磷酸,葡萄糖6-磷酸,和腺苷)通过使用综合蛋白质组学-代谢组学分析来鉴定导致细胞稳态失调的,这突出了细胞不稳定性的增加。这些发现阐明了培养连续传代时的一些重要分子过程,这有助于体外生物学的整体观点,重点是心血管细胞的复制衰老。
Long-term cell culture is an important biological approach but is also characterized by degeneration in cellular morphology, proliferation rate, and function. To explore this phenomenon in a systematic way, we conducted an integrative proteomics-metabolomics measurement of two cardiovascular cell lines of AC16 and HUVECs. The 18th culturing passages, i.e., G18, showed as the turning points by cell metabolism profiles, in which the metabolomic changes demonstrated the dysfunction of energy, amino acid, and ribonucleotide metabolism metabolic pathways. Although active protein networks showed mitochondria abundance AC16 and oxidative/nitrative sensitive HUVECs indicated the different degeneration patterns, the G18 and G30 proteomics evidenced the senescence by processes of signal transduction, signaling by interleukins, programmed cell death, cellular responses to stimuli, cell cycle, mRNA splicing, and translation. Some crucial proteins (RPS8, HNRNPR, SOD2, LMNB1, PSMA1, DECR1, GOT2, OGDH, PNP, CBS, ATIC, and IMPDH2) and metabolites (L-glutamic acid, guanine, citric acid, guanosine, guanosine diphosphate, glucose 6-phosphate, and adenosine) that contributed to the dysregulation of cellular homeostasis are identified by using the integrative proteomic-metabolomic analysis, which highlighted the increased cellular instability. These findings illuminate some vital molecular processes when culturing serial passages, which contribute holistic viewpoints of in vitro biology with emphasis on the replicative senescence of cardiovascular cells.