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Proteomic fingerprinting of N-linked glycoproteins involved in hepatocellular carcinoma
MA Jin QI Yi-jun* LIU Rui-min WANG Ming ZHANG Tian ZHU Han MA Yuan-fang
Acta Anatomica Sinica ›› 2014, Vol. 45 ›› Issue (4) : 493-499.
Proteomic fingerprinting of N-linked glycoproteins involved in hepatocellular carcinoma
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 potentialin 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 vitroinvasive 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.
Hepatocellular carcinoma / Glycoproteins / Cathepsin D / Haptoglobin / Human liver carboxylesterase1 / Two-dimensional electrophoresis / Mass spectrometry / Human
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the National Natural Science Foundation of China
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