title: Quantitative proteome analysis of colorectal cancer-related differential proteins.
abstract: OBJECTIVE
  To evaluate a new strategy for profiling proteomic changes in colorectal cancer (CRC).
  
  
  METHODS
  We used laser capture microdissection (LCM) to obtain cells from 20 CRC and paired normal mucosal tissues. The differential proteins between the microdissected tumor cells and normal mucosa epithelia were analyzed by acetylation stable isotopic labeling coupled with L linear ion trap Fourier transform ion cyclotron resonance mass spectrometry (LTQ-FT MS). Western blotting was used to assess the differential expression of proteins. We used bioinformatics tools for cluster and ingenuity pathway analysis of the differential proteins.
  
  
  RESULTS
  In total, 798 confident proteins were quantified and 137 proteins were differentially expressed by at least twofold, including 67 that were upregulated and 70 that were downregulated in cancer. Two differential proteins, solute carrier family 12 member 2 (SLC12A2) and Ras-related protein Rab-10, were validated by Western blotting, and the results were consistent with acetylation stable isotopic labeling analysis. According to gene ontology analysis, CRC-related differential proteins covered a wide range of subcellular locations and were involved in many biological processes. According to ingenuity pathway analysis of the differential proteins, the most relevant canonical pathway associated with CRC was the 14-3-3-mediated signaling pathway, and seven reliable functional networks including cellular growth and proliferation, amino acid metabolism, inflammatory response, embryonic development, carbohydrate metabolism, cellular assembly and organization, and cell morphology were obtained.
  
  
  CONCLUSIONS
  Combination of LCM, acetylation stable isotopic labeling analysis and LTQ-FT MS is effective for profiling proteomic changes in CRC cells.
authors.affiliation: Beijing Key Lab for Therapeutic Cancer Vaccines, Capital Medical University Cancer Center, Beijing Shijitan Hospital, The Capital Medical University, Beijing, China.
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Beijing Key Lab for Therapeutic Cancer Vaccines, Capital Medical University Cancer Center, Beijing Shijitan Hospital, The Capital Medical University, Beijing, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Gastrointestinal Surgery, Peking University People's Hospital, Peking University, Beijing, China.
authors.forename: Yingjiang
authors.initials: Y
authors.lastname: Ye
authors.affiliation: Department of Gastrointestinal Surgery, Peking University People's Hospital, Peking University, Beijing, China.
authors.forename: Danhua
authors.initials: D
authors.lastname: Shen
authors.affiliation: Department of Gastrointestinal Surgery, Peking University People's Hospital, Peking University, Beijing, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Beijing Key Lab for Therapeutic Cancer Vaccines, Capital Medical University Cancer Center, Beijing Shijitan Hospital, The Capital Medical University, Beijing, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Huang
authors.affiliation: Beijing Key Lab for Therapeutic Cancer Vaccines, Capital Medical University Cancer Center, Beijing Shijitan Hospital, The Capital Medical University, Beijing, China.
authors.forename: Sha
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Gastrointestinal Surgery, Peking University People's Hospital, Peking University, Beijing, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Wang
authors.affiliation: Beijing Key Lab for Therapeutic Cancer Vaccines, Capital Medical University Cancer Center, Beijing Shijitan Hospital, The Capital Medical University, Beijing, China. jun.ren@duke.edu.
authors.forename: Jun
authors.initials: J
authors.lastname: Ren
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015179:Colorectal Neoplasms / Q000378:metabolism*; D015870:Gene Expression; D063990:Gene Ontology; D006801:Humans; D053858:Metabolic Networks and Pathways; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D014408:Biomarkers, Tumor; D020543:Proteome
keywords: Acetylation stable isotopic labeling; Colorectal cancer; Laser capture microdissection; Quantitative proteomics