SILAC-based quantitative proteomic approach to identify potential biomarkers from the esophageal squamous cell carcinoma secretome Academic Article uri icon

abstract

  • The identification of secreted proteins that are differentially expressed between non-neoplastic and esophageal squamous cell carcinoma (ESCC) cells can provide potential biomarkers of ESCC. We used a SILAC-based quantitative proteomic approach to compare the secretome of ESCC cells with that of non-neoplastic esophageal squamous epithelial cells. Proteins were resolved by SDS-PAGE, and tandem mass spectrometry analysis (LC-MS/MS) of in-gel trypsin-digested peptides was carried out on a high-accuracy qTOF mass spectrometer. In total, we identified 441 proteins in the combined secretomes, including 120 proteins with > 2-fold upregulation in the ESCC secretome vs. that of non-neoplastic esophageal squamous epithelial cells. In this study, several potential protein biomarkers previously known to be increased in ESCC including matrix metalloproteinase 1, transferrin receptor, and transforming growth factor beta-induced 68 kDa were identified as overexpressed in the ESCC-derived secretome. In addition, we identified several novel proteins that have not been previously reported to be associated with ESCC. Among the novel candidate proteins identified, protein disulfide isomerase family a member 3 (PDIA3), GDP dissociation inhibitor 2 (GDI2), and lectin galactoside binding soluble 3 binding protein (LGALS3BP) were further validated by immunoblot analysis and immunohistochemical labeling using tissue microarrays. This tissue microarray analysis showed overexpression of protein disulfide isomerase family a member 3, GDP dissociation inhibitor 2, and lectin galactoside binding soluble 3 binding protein in 93%, 93% and 87% of 137 ESCC cases, respectively. Hence, we conclude that these potential biomarkers are excellent candidates for further evaluation to test their role and efficacy in the early detection of ESCC.

authors

  • Kashyap, Manoj Kumar
  • Harsha, HC
  • Renuse, Santosh
  • Pawar, Harsh
  • Sahasrabuddhe, Nandini A
  • Kim, Min-Sik
  • Marimuthu, Arivusudar
  • Keerthikumar, Shivakumar
  • Muthusamy, Babylakshmi
  • Kandasamy, Kumaran
  • Subbannayya, Yashwanth
  • Prasad, Thottethodi Subrahmanya Keshava
  • Mahmood, Riaz
  • Chaerkady, Raghothama
  • Meltzer, Stephen J
  • Kumar, Rekha V
  • Rustgi, Anil K
  • Pandey, Akhilesh

publication date

  • October 15, 2010

has subject area