TY - JOUR
T1 - Proteomic interrogation of androgen action in prostate cancer cells reveals roles of aminoacyl tRNA synthetases
AU - Vellaichamy, Adaikkalam
AU - Sreekumar, Arun
AU - Strahler, John R.
AU - Rajendiran, Theckelnaycke
AU - Yu, Jindan
AU - Varambally, Sooryanarayana
AU - Li, Yong
AU - Omenn, Gilbert S.
AU - Chinnaiyan, Arul M.
AU - Nesvizhskii, Alexey I.
N1 - Funding Information:
Synthetic androgen R1881 (methyltrienolone) was purchased from Perkin Elmer (Boston, MA). A 10 mM R1881 stock was prepared in 100% ethanol and stored in −20°C. RPMI 1640 medium and fetal bovine serum was purchased from Invitrogen (Carlsbad, CA). Sequencing grade modified trypsin was from Promega (Madison, WI). Sample preparation reagents formic acid, trifluoroacetic acid, and HPLC solvents were from Fisher Scientific (Waltham, MA). Mass spectrometriy grade water was from Burdick and Jackson (Muskegon, MI, USA). Antibodies used were: KLK3 (kallikrein 3, rabbit polyclonal, DakoCytomation Carpinteria, CA ), NDRG1 (N-myc downstream regulated 1, goat polyclonal, Santa Cruz Biotechnology Inc., Santa Cruz, CA.), FASN (fatty acid synthase, mouse monoclonal, BD Transduction Labs, Franklin Lakes, NJ), FKBP5 (FK506 binding protein 5, mouse monoclonal, BD Transduction Labs), AR (mouse monoclonal-AR441, Lab Vision, Fremont, CA), GARS (glycyl-tRNA synthetase, rabbit polyclonal, Abcam, Cambridge, MA), WARS (tryptophanyl-tRNA synthetase, rabbit polyclonal, Abcam), KARS (lysyl-tRNA synthetase, rabbit polyclonal, Bethyl Laboratories, Montgomery, TX), β-tubulin (rabbit polyclonal, Santa Cruz), β-actin (rabbit polyclonal, Cell Signaling Technology, Inc., Danvers, MA). LNCaP cells were purchased from American Type Culture Collection (University Boulevard, Manassas, VA). Prostate specimens comprising adjacent normal, localized prostate cancer, and metastasis tissue utilized for immunoblot experiments were from the University of Michigan Rapid Autopsy Program which is part of University of Michigan Prostate Cancer Specialized Program of Research Excellence (SPORE) tissue core.
PY - 2009/9/18
Y1 - 2009/9/18
N2 - Prostate cancer remains the most common malignancy among men in United States, and there is no remedy currently available for the advanced stage hormone-refractory cancer. This is partly due to the incomplete understanding of androgen-regulated proteins and their encoded functions. Whole-cell proteomes of androgen-starved and androgen-treated LNCaP cells were analyzed by semi-quantitative MudPIT ESI- ion trap MS/ MS and quantitative iTRAQ MALDI- TOF MS/MS platforms, with identification of more than 1300 high-confidence proteins. An enrichment-based pathway mapping of the androgen-regulated proteomic data sets revealed a significant dysregulation of aminoacyl tRNA synthetases, indicating an increase in protein biosynthesis- a hallmark during prostate cancer progression. This observation is supported by immunoblot and transcript data from LNCaP cells, and prostate cancer tissue. Thus, data derived from multiple proteomics platforms and transcript data coupled with informatics analysis provides a deeper insight into the functional consequences of androgen action in prostate cancer.
AB - Prostate cancer remains the most common malignancy among men in United States, and there is no remedy currently available for the advanced stage hormone-refractory cancer. This is partly due to the incomplete understanding of androgen-regulated proteins and their encoded functions. Whole-cell proteomes of androgen-starved and androgen-treated LNCaP cells were analyzed by semi-quantitative MudPIT ESI- ion trap MS/ MS and quantitative iTRAQ MALDI- TOF MS/MS platforms, with identification of more than 1300 high-confidence proteins. An enrichment-based pathway mapping of the androgen-regulated proteomic data sets revealed a significant dysregulation of aminoacyl tRNA synthetases, indicating an increase in protein biosynthesis- a hallmark during prostate cancer progression. This observation is supported by immunoblot and transcript data from LNCaP cells, and prostate cancer tissue. Thus, data derived from multiple proteomics platforms and transcript data coupled with informatics analysis provides a deeper insight into the functional consequences of androgen action in prostate cancer.
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U2 - 10.1371/journal.pone.0007075
DO - 10.1371/journal.pone.0007075
M3 - Article
C2 - 19763266
AN - SCOPUS:70349446266
SN - 1932-6203
VL - 4
JO - PloS one
JF - PloS one
IS - 9
M1 - e7075
ER -
