Abstract
The aim of this study was to identify novel biomarkers for the diagnosis of, and potential therapeutic targets for, hepatocellular carcinoma (HCC). Multilectin affinity chromatography was used to enrich N-linked glycoproteins from nontumorous liver and HCC tissues followed by 2DE and protein identification by MS. Twenty-eight differentially expressed proteins were identified. Western blotting validated consistently lower concentrations of human liver carboxylesterase 1 and haptoglobin, and higher concentration of procathepsin D (pCD) in HCC tissues. Knockdown of cathepsin D (CD) expression mediated by siRNA significantly inhibited the in vitro invasion of two HCC cell lines, SNU449 and SNU473, which normally secrete high-levels of CD. Prefractionation using individual lectins demonstrated an elevation in ConA-binding glycoforms of proCD and CD in HCC tissues. In the serum of HCC patients, "ConA-binding proCD" (ConA-pCD) is significantly increased in concentration and this increase is comprised of several distinct upregulated acidic isoforms (pI 4.5-5.5). Receiver operating characteristic analysis showed that the sensitivity and specificity of serum ConA-pCD for HCC diagnosis were 85% and 80%, respectively. This is the first report that serum ConA-pCD is increased significantly in HCC and is potentially useful as a serological biomarker for diagnosis of HCC.
Original language | English |
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Pages (from-to) | 186-195 |
Number of pages | 10 |
Journal | Proteomics |
Volume | 14 |
Issue number | 2-3 |
Early online date | 20 Nov 2013 |
DOIs | |
Publication status | Published - Feb 2014 |
Keywords
- Adult
- Aged
- Aged, 80 and over
- Biomarkers, Tumor
- Blotting, Western
- Carboxylic Ester Hydrolases
- Carcinoma, Hepatocellular
- Cathepsin D
- Cell Line, Tumor
- Concanavalin A
- Enzyme Precursors
- Female
- Gene Expression Regulation, Neoplastic
- Glycoproteins
- Haptoglobins
- Humans
- Liver
- Liver Neoplasms
- Male
- Middle Aged
- Proteomics
- RNA Interference
- RNA, Small Interfering