Proteomic fingerprinting of N-linked glycoproteins involved in hepatocellular carcinoma

doi:10.3969/j.issn.0529-1356.2014.04.010

Abstract

Abstract:

Objective To identify differentially expressed N-linked glycoproteins between hepatocellular carcinoma (HCC) and adjacent non-tumorous liver tissues.
Methods N-linked glycoproteome was extracted by multi-lectin affinity chromatography comprising concanavalin A (ConA), lentil lectin (LCH), and snowdrop lectin (GNA) and subsequently subjected to two-dimensional electrophoresis (2DE) and mass spectrometry(MS) for identification of differential glycoproteins between 10 pairs of HCC and adjacent non-cancer tissue. Western blotting was used to verify different expression of human liver carboxylesterase1 (hCE1), haptoglobin (HP）and cathepsin D (CD). Invasion potentialin vitro was examined after si-RNA mediated CD gene scilencing. Results LC-ESI-MS/MS identified a total of 28 differentially expressed glycoproteins (14 up-regulation and 14 down-regulated). Western blotting detected consistent down-regulation of hCE1 and HP, and up-regulation of pro-cathepsin D (pCD) in HCC. Up-regulation of ConA-binding CD (ConA-CD), however, was verified in HCC only after ConA-CD enrichment by ConA chromatography. Down-regulation of CD expression mediated by CD-siRNA markedly inhibited the in vitroinvasive potential of SNU449 and SNU473.
Conclusion Dysregulation of HP, hCE1 expression and alteration of glycans linked to CD may play crucial roles in pathogenesis of HCC.

Key words: Hepatocellular carcinoma, Glycoproteins, Cathepsin D, Haptoglobin, Human liver carboxylesterase1, Two-dimensional electrophoresis, Mass spectrometry, Human

MA Jin QI Yi-jun* LIU Rui-min WANG Ming ZHANG Tian ZHU Han MA Yuan-fang. Proteomic fingerprinting of N-linked glycoproteins involved in hepatocellular carcinoma[J]. AAS, 2014, 45(4): 493-499.

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