Format

Send to

Choose Destination
J Proteome Res. 2018 Sep 7;17(9):2963-2977. doi: 10.1021/acs.jproteome.8b00135. Epub 2018 Aug 3.

Proteomic Analysis of Charcoal-Stripped Fetal Bovine Serum Reveals Changes in the Insulin-like Growth Factor Signaling Pathway.

Author information

1
Department of Pharmaceutical Sciences , State University of New York at Buffalo , 285 Kapoor Hall , Buffalo , New York 14260 , United States.
2
New York State Center of Excellence in Bioinformatics and Life Sciences , 701 Ellicott Street , Buffalo , New York 14203 , United States.
3
Department of Urology, Roswell Park Comprehensive Cancer Center , Elm and Carlton Streets , Buffalo , New York 14263 , United States.
4
Department of Pathology, Roswell Park Comprehensive Cancer Center , Elm and Carlton Streets , Buffalo , New York 14263 , United States.
5
Department of Pharmacy, Changzheng Hospital , Second Military Medical University , 415 Fengyang Road , Shanghai 200003 , China.
6
College of Mechanical Engineering , Dongguan University of Technology , 1 Daxue Road , Dongguan , Guangdong 523808 , China.
7
Department of Cancer Prevention and Control , Roswell Park Comprehensive Cancer Center , Elm and Carlton Streets , Buffalo , New York 14263 , United States.

Abstract

Charcoal-stripped fetal bovine serum (CS-FBS) is commonly used to study androgen responsiveness and androgen metabolism in cultured prostate cancer (CaP) cells. Switching CaP cells from FBS to CS-FBS may reduce the activity of androgen receptor (AR), inhibit cell proliferation, or modulate intracellular androgen metabolism. The removal of proteins by charcoal stripping may cause changes in biological functions and has not yet been investigated. Here we profiled proteins in FBS and CS-FBS using an ion-current-based quantitative platform consisting of reproducible surfactant-aided precipitation/on-pellet digestion, long-column nanoliquid chromatography separation, and ion-current-based analysis. A total of 143 proteins were identified in FBS, among which 14 proteins including insulin-like growth factor 2 (IGF-2) and IGF binding protein (IGFBP)-2 and -6 were reduced in CS-FBS. IGF-1 receptor (IGF1R) and insulin receptor were sensitized to IGFs in CS-FBS. IGF-1 and IGF-2 stimulation fully compensated for the loss of AR activity to maintain cell growth in CS-FBS. Endogenous production of IGF and IGFBPs was verified in CaP cells and clinical CaP specimens. This study provided the most comprehensive protein profiles of FBS and CS-FBS and offered an opportunity to identify new protein regulators and signaling pathways that regulate AR activity, androgen metabolism, and proliferation of CaP cells.

KEYWORDS:

IGF-1; cell culture; charcoal stripping; fetal bovine serum; growth factors; medium; phosphorylation; prostate cancer; proteomics; serum

PMID:
30014700
DOI:
10.1021/acs.jproteome.8b00135
[Indexed for MEDLINE]

Publication types, MeSH terms, Substances, Grant support

Publication types

MeSH terms

Substances

Grant support

Supplemental Content

Full text links

Icon for American Chemical Society
Loading ...
Support Center