Trypsin-induced proteome alteration during cell subculture in mammalian cells.
Trypsin-induced proteome alteration during cell subculture in mammalian cells.
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DOI：
10.1186/1423-0127-17-36
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发表时间：
2010-05-11
影响因子：
11
通讯作者：
Chan HL
中科院分区：
文献类型：
--
作者：
Huang HL;Hsing HW;Lai TC;Chen YW;Lee TR;Chan HT;Lyu PC;Wu CL;Lu YC;Lin ST;Lin CW;Lai CH;Chang HT;Chou HC;Chan HL
一旦培养的细胞达到汇合，就必须对细胞进行传代培养。为此，胰蛋白酶经常用于从基质中分离粘附细胞。然而，由于胰蛋白酶的蛋白水解活性，细胞表面蛋白通常被切割，这导致细胞功能失调。在这项研究中，一式三份的2D-DIGE策略已经进行了监测胰蛋白酶诱导的蛋白质组改变。通过MALDI-TOF MS鉴定差异表达点，并通过免疫印迹进行验证。发现36种蛋白质在胰蛋白酶处理的细胞中差异表达，已知调节细胞代谢、生长调节、线粒体电子传递和细胞粘附的蛋白质在胰蛋白酶处理后下调，调节细胞凋亡的蛋白质在胰蛋白酶处理后上调。进一步研究发现，胰蛋白酶消化后bcl-2表达下调，p53和p21表达上调。总之，这是第一份报告，使用蛋白质组学的方法，彻底研究胰蛋白酶诱导的细胞生理变化，并提供研究人员在执行他们的实验设计。
It is essential to subculture the cells once cultured cells reach confluence. For this, trypsin is frequently applied to dissociate adhesive cells from the substratum. However, due to the proteolytic activity of trypsin, cell surface proteins are often cleaved, which leads to dysregulation of the cell functions. In this study, a triplicate 2D-DIGE strategy has been performed to monitor trypsin-induced proteome alterations. The differentially expressed spots were identified by MALDI-TOF MS and validated by immunoblotting. 36 proteins are found to be differentially expressed in cells treated with trypsin, and proteins that are known to regulate cell metabolism, growth regulation, mitochondrial electron transportation and cell adhesion are down-regulated and proteins that regulate cell apoptosis are up-regulated after trypsin treatment. Further study shows that bcl-2 is down-regulated, p53 and p21 are both up-regulated after trypsinization. In summary, this is the first report that uses the proteomic approach to thoroughly study trypsin-induced cell physiological changes and provides researchers in carrying out their experimental design.
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影响因子：
2.8
作者：
Lee, JW;Soung, YH;Lee, SH
通讯作者：
Lee, SH
影响因子：
3.9
作者：
Tsujimoto, Y;Shimizu, S
通讯作者：
Shimizu, S
影响因子：
3.4
作者：
Jang, JH;Hanash, S
通讯作者：
Hanash, S
影响因子：
4.4
作者：
Lai, Tzu-Chia;Chou, Hsiu-Chuan;Chan, Hong-Lin
通讯作者：
Chan, Hong-Lin
影响因子：
3.4
作者：
Chan, HL;Gharbi, S;Timms, JF
通讯作者：
Timms, JF