Faculty Collaboration Database

Quantitative proteomic profiling studies of pancreatic cancer stem cells. J Proteome Res 2010 Jul 02;9(7):3394-402

Date

05/22/2010

Pubmed ID

20486718

Pubmed Central ID

PMC2906393

DOI

10.1021/pr100231m

Scopus ID

2-s2.0-77954362750 (requires institutional sign-in at Scopus site)   27 Citations

Abstract

Analyzing subpopulations of tumor cells in tissue is a challenging subject in proteomic studies. Pancreatic cancer stem cells (CSCs) are such a group of cells that only constitute 0.2-0.8% of the total tumor cells but have been found to be the origin of pancreatic cancer carcinogenesis and metastasis. Global proteome profiling of pancreatic CSCs from xenograft tumors in mice is a promising way to unveil the molecular machinery underlying the signaling pathways. However, the extremely low availability of pancreatic tissue CSCs (around 10,000 cells per xenograft tumor or patient sample) has limited the utilization of currently standard proteomic approaches which do not work effectively with such a small amount of material. Herein, we describe the profiling of the proteome of pancreatic CSCs using a capillary scale shotgun technique by coupling offline capillary isoelectric focusing(cIEF) with nano reversed phase liquid chromatography(RPLC) followed by spectral counting peptide quantification. A whole cell lysate from 10,000 cells which corresponds to approximately 1 microg of protein material is equally divided for three repeated cIEF separations where around 300 ng of peptide material is used in each run. In comparison with a nontumorigenic tumor cell sample, among 1159 distinct proteins identified with FDR less than 0.2%, 169 differentially expressed proteins are identified after multiple testing corrections where 24% of the proteins are upregulated in the CSCs group. Ingenuity Pathway analysis of these differential expression signatures further suggests significant involvement of signaling pathways related to apoptosis, cell proliferation, inflammation, and metastasis.

Author List

Dai L, Li C, Shedden KA, Lee CJ, Li C, Quoc H, Simeone DM, Lubman DM

MESH terms used to index this publication - Major topics in bold

Animals
Chromatography, Reverse-Phase
Cluster Analysis
Gene Expression Profiling
Humans
Isoelectric Focusing
Male
Mice
Mice, Inbred NOD
Mice, SCID
Neoplastic Stem Cells
Pancreatic Neoplasms
Proteins
Proteomics
Signal Transduction
Xenograft Model Antitumor Assays