title: Trypsin-induced proteome alteration during cell subculture in mammalian cells.
abstract: 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.
authors.affiliation: Institute of Bioinformatics and Structural Biology & Department of Life Sciences, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Hsiang-Ling
authors.initials: HL
authors.lastname: Huang
authors.forename: Hsiang-Wei
authors.initials: HW
authors.lastname: Hsing
authors.forename: Tzu-Chia
authors.initials: TC
authors.lastname: Lai
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Chen
authors.forename: Tian-Ren
authors.initials: TR
authors.lastname: Lee
authors.forename: Hsin-Tsu
authors.initials: HT
authors.lastname: Chan
authors.forename: Ping-Chiang
authors.initials: PC
authors.lastname: Lyu
authors.forename: Chieh-Lin
authors.initials: CL
authors.lastname: Wu
authors.forename: Ying-Chieh
authors.initials: YC
authors.lastname: Lu
authors.forename: Szu-Ting
authors.initials: ST
authors.lastname: Lin
authors.forename: Cheng-Wen
authors.initials: CW
authors.lastname: Lin
authors.forename: Chih-Ho
authors.initials: CH
authors.lastname: Lai
authors.forename: Hao-Teng
authors.initials: HT
authors.lastname: Chang
authors.forename: Hsiu-Chuan
authors.initials: HC
authors.lastname: Chou
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Chan
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005260:Female; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000378:metabolism; D020543:Proteome / Q000187:drug effects* / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000378:metabolism
chemical_list: C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D009363:Neoplasm Proteins; D020543:Proteome; D019253:Proto-Oncogene Proteins c-bcl-2; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D014357:Trypsin
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