The Cell Line Secretome, a Suitable Tool for Investigating Proteins Released in Vivo by Tumors: Application to the Study of p53-Modulated Proteins Secreted in Lung Cancer Cells

被引:44
作者
Chenau, Jerome [3 ]
Michelland, Sylvie [1 ,3 ]
de Fraipont, Florence [2 ,3 ]
Josserand, Veronique [3 ]
Coll, Jean-Luc [3 ]
Favrot, Marie-Christine [1 ,3 ]
Seve, Michel [1 ,3 ]
机构
[1] CHU Grenoble, Ctr Innovat Biol, F-38043 Grenoble 09, France
[2] CHU Grenoble, Dept Biol Integree, F-38043 Grenoble 09, France
[3] Univ Grenoble 1, INSERM, Inst Albert Bonniot, U823, Grenoble, France
关键词
Proteomics; iTRAQ; lung cancer; p53; secretome; xenograft mouse model; WILD-TYPE P53; FIBROBLAST-GROWTH-FACTOR; HUMAN GLIOMA-CELLS; GENE-EXPRESSION; OFFGEL ELECTROPHORESIS; P53-INDUCED APOPTOSIS; SUPPRESSOR GENE; SIGNAL PEPTIDES; BINDING PROTEIN; UP-REGULATION;
D O I
10.1021/pr900383g
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Malignant processes such as metastasis, invasion, or angiogenesis are tightly dependent on the composition of the extracellular medium, which is itself affected by the release of proteins by the tumor cells. p53, a major tumor suppressor protein very frequently mutated and/or inactivated in cancer cells, is known to modulate the release of proteins by the tumor cells; however, while p53-modulated intracellular proteins have been extensively studied, little is known concerning their extracellular counterparts. Here, we characterized the p53-dependent secretome of a lung tumor model in vitro (H358 human nonsmall cell lung adenocarcinoma cell line with a homozygous deletion of p53) and demonstrate that the modulation of exported proteins can also be detected in vivo in the plasma of tumor-bearing mice. We used a clone of H358, stably transfected with a tetracycline-inducible wildtype p53-expressing vector. With the use of iTRAQ labeling and LC-MALDI-MS/MS analysis, we identified 909 proteins released in vitro by the cells, among which 91 are p53-modulated. Three proteins (GDF-15, FGF-19, and VEGF) were also investigated in H358/TetOn/p53 xenograft mice. The ELISA dosage on total tumor protein extracts confirmed the influence of p53 on the release of these proteins in vivo. Moreover, the GDF-15 concentration was measured in the plasma and its p53-dependent modulation was confirmed. To our knowledge, this is the first report establishing that the in vitro cell line secretome is reliable and reflects the extracellular release of proteins from tumor cells in vivo and could be used to identify putative tumor markers.
引用
收藏
页码:4579 / 4591
页数:13
相关论文
共 93 条
[1]   The fibroblast growth factor-binding protein FGF-BP [J].
Abuharbeid, Shaker ;
Czubayko, Frank ;
Aigner, Achim .
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 2006, 38 (09) :1463-1468
[2]  
Atencio IA, 2000, CELL GROWTH DIFFER, V11, P247
[3]   Feature-based prediction of non-classical and leaderless protein secretion [J].
Bendtsen, JD ;
Jensen, LJ ;
Blom, N ;
von Heijne, G ;
Brunak, S .
PROTEIN ENGINEERING DESIGN & SELECTION, 2004, 17 (04) :349-356
[4]   Improved prediction of signal peptides: SignalP 3.0 [J].
Bendtsen, JD ;
Nielsen, H ;
von Heijne, G ;
Brunak, S .
JOURNAL OF MOLECULAR BIOLOGY, 2004, 340 (04) :783-795
[5]   The emerging roles of human tissue kallikreins in cancer [J].
Borgoño, CA ;
Diamandis, EP .
NATURE REVIEWS CANCER, 2004, 4 (11) :876-890
[6]  
Brambilla E, 1996, AM J PATHOL, V149, P1941
[7]   INDUCTION OF THE GROWTH INHIBITOR IGF-BINDING PROTEIN-3 BY P53 [J].
BUCKBINDER, L ;
TALBOTT, R ;
VELASCOMIGUEL, S ;
TAKENAKA, I ;
FAHA, B ;
SEIZINGER, BR ;
KLEY, N .
NATURE, 1995, 377 (6550) :646-649
[8]   Peptides OFFGEL electrophoresis: a suitable pre-analytical step for complex eukaryotic samples fractionation compatible with quantitative iTRAQ labeling [J].
Chenau, Jerome ;
Michelland, Sylvie ;
Sidibe, Jonathan ;
Seve, Michel .
PROTEOME SCIENCE, 2008, 6 (1)
[9]   Antitumor activity of bax and p53 naked gene transfer in lung cancer:: In vitro and in vivo analysis [J].
Coll, JL ;
Negoescu, A ;
Louis, N ;
Sachs, L ;
Tenaud, C ;
Girardot, V ;
Demeinex, B ;
Brambilla, E ;
Brambilla, C ;
Favrot, M .
HUMAN GENE THERAPY, 1998, 9 (14) :2063-2074
[10]   wt p53 dependent expression of a membrane-associated isoform of adenylate kinase [J].
Collavin, L ;
Lazarevic, D ;
Utrera, R ;
Marzinotto, S ;
Monte, M ;
Schneider, C .
ONCOGENE, 1999, 18 (43) :5879-5888