Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

iTRAQ-facilitated Proteomic analysis of human prostate cancer cells identifies proteins associated with progression

Glen, Adam, Gan, Chee S., Hamdy, Freddie C., Eaton, Colby L., Cross, Simon S., Catto, James W. F., Wright, Phillip C. and Rehman, Ishtiaq 2008. iTRAQ-facilitated Proteomic analysis of human prostate cancer cells identifies proteins associated with progression. Journal of Proteome Research 7 (3) , pp. 897-907. 10.1021/pr070378x

Full text not available from this repository.


The unpredictable behavior of prostate cancer presents a major clinical challenge during patient management. In order to gain an insight into the molecular mechanisms associated with prostate cancer progression, we employed the shot-gun proteomic approach of isobaric tags for relative and absolute quantitation (iTRAQ), followed by 2D-LC-MS/MS, using the poorly metastatic LNCaP cell line and its highly metastatic variant LNCaP-LN3 cell line as a model. A total number of 280 unique proteins were identified (≥95% confidence), and relative expression data was obtained for 176 of these. Ten proteins were found to be significantly up-regulated (≥1.50 fold), while 4 proteins were significantly down-regulated (≥−1.50 fold), in LNCaP-LN3 cells. Differential expression of brain creatine kinase (CKBB), soluble catechol- O-methyltransferase (S-COMT), tumor rejection antigen (gp96), and glucose regulated protein, 78 kDa (grp78), was confirmed by Western blotting or independent 2D-PAGE analysis. Additionally, iTRAQ analysis identified absence of the lactate dehydrogenase-B (LDH-B) subunit in LNCaP-LN3 cells, confirming our published data. The clinical relevance of gp96 was assessed by immunohistochemistry using prostate tissues from benign ( n = 95), malignant ( n = 66), and metastatic cases ( n = 3). Benign epithelium showed absent/weak gp96 expression in the basal cells, in contrast to the moderate/strong expression seen in malignant epithelium. Furthermore, there was a statistically significant difference in the intensity of gp96 expression between benign and malignant cases ( p < 0.0005, Mann–Whitney U). Our study is the first to report the application of iTRAQ technology and its potential for the global proteomic profiling of prostate cancer cells, including the identification of absent protein expression.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Dentistry
Subjects: R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
R Medicine > RK Dentistry
Uncontrolled Keywords: benign hyperplasia; shot-gun proteomics; 2D-PAGE; immunochemistry; tumor rejection antigen (gp96)
Publisher: American Chemical Society
ISSN: 1535-3893
Last Modified: 10 Oct 2017 14:28

Citation Data

Cited 89 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item