- 作者: Hsiang-Ling Huang; Hsiang-Wei Hsing; Tzu-Chia Lai; Yi-Wen Chen; Tian-Ren Lee; Hsin-Tsu Chan; Ping-Chiang Lyu; Chieh-Lin Wu; Ying-Chieh Lu; Szu-Ting Lin; Cheng-Wen Lin; Chih-Ho Lai; Hao-Teng Chang; Hsiu-Chuan Chou; Hong-Lin Chan
- 作者服務機構: Institute of Bioinformatics and Structural Biology & Department of Life Sciences, National Tsing Hua University, Taiwan, R.O.C.
- 中文摘要: --
- 英文摘要:
Background : 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.
Methods : 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.
Results : 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.
Conclusions : 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. - 中文關鍵字: --
- 英文關鍵字: --