Modified serum-guided immunoblotting for differential proteomic study of prostate cancer.
- Author:
Xiao-bo ZHANG
1
;
Zheng-yan TANG
;
Xiong-bin ZU
;
Lin QI
;
Jing-de RUAN
Author Information
- Publication Type:Journal Article
- MeSH: Biomarkers, Tumor; blood; Blotting, Western; Humans; Immunoblotting; methods; Male; Mass Spectrometry; Prostatic Neoplasms; genetics; metabolism; Proteomics
- From: National Journal of Andrology 2010;16(5):438-444
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo search for a new method of screening for molecular targets for androgen-dependent prostate cancer.
METHODSWe collected tissue samples and paired serum samples from 3 cases of androgen-dependent prostate cancer (ADPC) treated by surgical resection, and included another 3 samples of benign prostatic hyperplasia (BPH) tissue and normal human serum in the control group. The total proteins extracted were separated and transmembrane by two-dimensional gel electrophoresis, followed by hybridization with the sera of the patients with ADPC and those with hormone-independent prostate cancer (HIPC) as the primary antibodies. The differentially expressed proteins were compared by Western blot, analyzed by MALDI-TOF-MS mass spectrography, and verified by RT-PCR and Western blot following bioinformatic identification.
RESULTSThis modified method exhibited a significantly better effect in displaying differentially expressed proteins, by which 12 differentially expressed protein spots were identified, including Beclin1, glutathione S-transferase P (GSTP1-1), ZBTB7, dihydrodiol dehydrogenase 2 (DDH), enolase (ENO1), glucose-dependent insulin-releasing peptide receptor (GIPR), Mn-superoxide dismutase (MnSOD), phosphoglycerate mutase 1 (PGAM1), amino-peptidyl-prolyl cistrons isomerase (PPIA), and phospholipid-PE-binding protein (PEBP). The mRNA and protein expressions of Beclin1 were significantly down-regulated in androgen-dependent prostate cancer tissues.
CONCLUSIONThis modified serum-guided immunoblotting technique has provided a new method for clarifying the molecular mechanisms of the occurrence and progression of HIPC, in which Beclin1-mediated autophagy may play a key role.