title: Proteomic analysis of primary colon cancer-associated fibroblasts using the SELDI-ProteinChip platform.
abstract: OBJECTIVE
  Cancer-associated fibroblasts (CAFs) are one of the hallmarks of the cancer microenvironment. Recent evidence has indicated that CAFs are more competent in enhancing cancer cell growth and migration than normal fibroblasts. However, the unique protein expression of CAFs has not been fully elucidated. This study aims to investigate the characterizations of colon CAFs by comparing the differential protein expression between CAFs and normal fibroblasts.
  
  
  METHODS
  Primary fibroblasts were isolated from surgical specimen of human colon cancer and matched normal colonic tissue. Purity of the cell population was verified through immunostain analysis. Total cell lysates and conditioned media from each group of cells were extracted, and protein expression analysis was conducted using the surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) ProteinChip platform.
  
  
  RESULTS
  Most primary cells showed typical fibroblast-like features after two weeks. Increased proportion of α-smooth muscle actin-positive myofibroblasts was detected within the CAFs in four of the six pairs of primary cells. Fibroblast activation protein was weakly expressed in most cells without differences. Using SELDI-TOF-MS ProteinChip platform, four protein peaks mass over charge ratio (m/z) 1142, 3011, 4035, and 4945 were detected in the total cell lysates, and two protein peaks m/z 1368 and 1389 were detected in the conditioned media. The potential candidate proteins found in the Swiss-Prot database include morphogenetic neuropeptides, FMRFamide-related peptides, insulin-like growth factor II, thymosin β-4-like protein 3, and tight junction-associated protein 1.
  
  
  CONCLUSIONS
  Using the SELDI-ProteinChip platform, differential protein expressions were identified in colon CAFs compared with normal colonic stromal fibroblasts. The complex proteomic alternations in colon CAFs may play important roles related to the colon cancer microenvironment.
authors.affiliation: Cancer Institute, the Second Affiliated Hospital School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Zhan-Huai
authors.initials: ZH
authors.lastname: Wang
authors.forename: Ke-Feng
authors.initials: KF
authors.lastname: Ding
authors.forename: Jie-Kai
authors.initials: JK
authors.lastname: Yu
authors.forename: Xiao-Hui
authors.initials: XH
authors.lastname: Zhai
authors.forename: Shu-Qin
authors.initials: SQ
authors.lastname: Ruan
authors.forename: Shan-Wei
authors.initials: SW
authors.lastname: Wang
authors.forename: Yong-Liang
authors.initials: YL
authors.lastname: Zhu
authors.forename: Shu
authors.initials: S
authors.lastname: Zheng
authors.forename: Su-Zhan
authors.initials: SZ
authors.lastname: Zhang
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D018929:Cell Culture Techniques; D002469:Cell Separation; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000378:metabolism; D040081:Protein Array Analysis; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment / Q000502:physiology
chemical_list: D009363:Neoplasm Proteins