Identification and analysis of altered alpha1,6-fucosylated glycoproteins associated with hepatocellular carcinoma metastasis.
Tumor metastasis might be associated with the expression levels of cellular glycoproteins and the alteration of their glycan parts. In order to screen the aberrantly alpha1,6-fucosylated glycoproteins related to hepatocellular carcinoma (HCC) metastasis, a high-throughput glycomic approach which consisted of 2-DE, electronic transfer of proteins, lectin affinity blot and precipitation, and MALDI-TOF-MS/MS, was established. Lens culinaris agglutinin (LCA) affinity glycoprotein profiles of higher and lower metastatic HCC cell lines were compared and analyzed. Seven out of 34 identified glycoproteins were differentially displayed; they were cytokeratin 8 (CK8), annexin I, annexin II, heterogeneous nuclear ribonucleoprotein A/B, PDZ and LIM domain 1, RNA-binding motif protein 4, and poly(rC)-binding protein 1. On comparison with Hep3B, CK8 showed a higher affinity to Ricinus communis agglutinin 1 (RCA-I) and LCA, and annexin I presented a higher affinity to LCA and Con A by the lectin-binding assay. Furthermore, the up-regulation of CK8, annexin I, and annexin II were found by Western blot and immunofluorescence analysis in higher metastatic HCC cell lines. This implied that the alteration of CK8, annexin I, and annexin II both in their expression levels and their glycan parts might be related to metastatic ability, and play a critical role in the process of HCC metastasis.
Dai
Zhi
Z
Liver Cancer Institute and Zhongshan Hospital, Fudan University, Shanghai, PR China.
Liu
Yin-Kun
YK
Cui
Jie-Feng
JF
Shen
Hua-Li
HL
Chen
Jie
J
Sun
Rui-Xia
RX
Zhang
Yu
Y
Zhou
Xin-Wen
XW
Yang
Peng-Yuan
PY
Tang
Zhao-You
ZY
D017305:Annexin A1 / Q000032:analysis; D017306:Annexin A2 / Q000032:analysis; D014408:Biomarkers, Tumor / Q000032:analysis; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000737:chemistry* / Q000473:pathology; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D005643:Fucose / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis*; D006031:Glycosylation; D006801:Humans; D053533:Keratin-8 / Q000032:analysis; D037102:Lectins / Q000378:metabolism; D008113:Liver Neoplasms / Q000737:chemistry* / Q000473:pathology; D008970:Molecular Weight; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000032:analysis*; D010449:Peptide Mapping; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling
D017305:Annexin A1; D017306:Annexin A2; D014408:Biomarkers, Tumor; D006023:Glycoproteins; D053533:Keratin-8; D037102:Lectins; D009363:Neoplasm Proteins; D005643:Fucose