Comparison of GP73 Glycosylation in Human Hepatocellur Carcinoma Cell Lines with Different Metastatic Potential
ZHANG Shu1, JIANG Kai2, MAO Xiaoyi3, LIU Yinkun1,2
1. Key Laboratory of Carcinogenesis and Cancer Invasion of Ministry of Education; Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China;
2. Cancer Research Center of Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China;
3. School of Basic Medical Sciences, Fudan University, Shanghai 200032, China
Abstract：In this study, Golgi protein 73 (GP73) of hepatocelluler carcinoma (HCC) cell lines with different metastatic potential was purified by antibody affinity chromatography and the purification result was confirmed by LC-MS/MS, providing more proof for GP73 as a biomarker for HCC. Purified GP73 was then analyzed using western blot and lectin blot to obtain its glycosylation level in HCC cell lines with different metastatic potential. We found that glycan of GP73 can be recognized by ConA, AAL, LCA and PHA-E lectins, while the LCA and PHA-E-reactive fraction of GP73 was increased along with the increasing metastatic potential of HCC cell lines. The glycan level of GP73 was different in HCC cell lines with different metastatic potential, and the content of fucose (recognized by LCA) and bisecting structure (recognized by PHA-E) increased in agreement with the metastatic potential.
张舒, 江凯, 茅晓意, 刘银坤. 不同转移潜能人肝癌细胞系中GP73糖基化修饰水平比较[J]. 科技导报, 2014, 32(16): 15-19.
ZHANG Shu, JIANG Kai, MAO Xiaoyi, LIU Yinkun. Comparison of GP73 Glycosylation in Human Hepatocellur Carcinoma Cell Lines with Different Metastatic Potential. journal1, 2014, 32(16): 15-19.
 Marrero J A, Romano P R, Nikolaeva O, et al. GP73, a resident Golgi glycoprotein, is a novel serum marker for hepatocellular carcinoma[J]. Journal of Hepatology, 2005, 43(6): 1007-1012.
 Bachert C, Fimmel C, Linstedt A D. Endosomal trafficking and proprotein convertase cleavage of cis Golgi protein GP73 produces marker for hepatocellular carcinoma[J]. Traffic, 2007, 8(10): 1415-1423.
 Mao Y, Yang H, Xu H, et al. Golgi protein 73 (GOLPH2) is a valuable serum marker for hepatocellular carcinoma[J]. Gut, 59(12): 1687-1693.
 Drake R R, Schwegler E E, Malik G, et al. Lectin capture strategies combined with mass spectrometry for the discovery of serum glycoprotein biomarkers[J]. Molecular of Cell Proteomics, 2006, 5(10): 1957-1967.
 Hu L, Li L, Xie H, et al. The Golgi localization of GOLPH2 (GP73/ GOLM1) is determined by the transmembrane and cytoplamic sequences[J]. PLoS One, 2011, 6(11): e28207.
 Norton P A, Comunale M A, Krakover J, et al. N-linked glycosylation of the liver cancer biomarker GP73[J]. Journal of Cell Biochemistry, 2008, 104(1): 136-149.
 Wang M, Long R E, Comunale M A, et al. Novel fucosylated biomarkers for the early detection of hepatocellular carcinoma[J]. Cancer Epidemiology Biomarkers Prevention, 2009, 18(6): 1914-1921.
 Block T M, Comunale M A, Lowman M, et al. Use of targeted glycoproteomics to identify serum glycoproteins that correlate with liver cancer in woodchucks and humans[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(3): 779-784.
 Morelle W, Michalski J C, Analysis of protein glycosylation by mass spectrometry[J]. Nature Protocols, 2007, 2(7): 1585-1602.
 Marino K, Bones J, Kattla J J, et al. A systematic approach to protein glycosylation analysis: A path through the maze[J]. Nature Chemical Biology, 6(10): 713-723.
 Amos B, Lotan D, Lotan R. Increased fucosylation of high-molecularweight glycoproteins accompanies retinoic-acid-induced differentiation of F-9 embryonal carcinoma cells[J]. International Journal of Cancer, 1990, 46(1): 86-94.
 Nakagawa T, Miyoshi E, Yakushijin T, et al. Glycomic analysis of alpha-fetoprotein L3 in hepatoma cell lines and hepatocellular carcinoma patients[J]. Journal of Proteome Research, 2008, 7(6): 2222-2233.
 Yamashita K, Koide N, Endo T, et al. Altered glycosylation of serum transferrin of patients with hepatocellular carcinoma[J]. The Journal of Biological Chemistry, 1989, 264(5): 2415-2423.
 Nishihara S, Iwasaki H, Kaneko M, et al. Alpha1, 3-fucosyltransferase 9 (FUT9, Fuc-TIX) preferentially fucosylates the distal GlcNAc residue of polylactosamine chain while the other four alpha1, 3FUT members preferentially fucosylate the inner GlcNAc residue[J]. FEBS Letters, 1999, 462(3): 289-294.
 Orntoft T F, Vestergaard E M, Clinical aspects of altered glycosylation of glycoproteins in cancer[J]. Electrophoresis, 1999, 20(2): 362-371.
 Kim Y J, Varki A, Perspectives on the significance of altered glycosylation of glycoproteins in cancer[J]. Glycoconjugate Journal, 1997, 14(5): 569-576.
 Miyoshi E, Ihara Y, Hayashi N, et al. Transfection of Nacetylglucosaminyltransferase Ⅲ gene suppresses expression of hepatitis B virus in a human hepatoma cell line, HB611[J]. The Journal of Biological Chemistry, 1995, 270(47): 28311-28315.
 Kang S K, Chung T W, Lee J Y, et al. The hepatitis B virus X protein inhibits secretion of apolipoprotein B by enhancing the expression of N-acetylglucosaminyltransferase Ⅲ[J]. The Journal of Biological Chemistry, 2004, 279(27): 28106-28112.
 董芳, 肖凡, 魏红山. 平分型GlcNAc糖基化修饰的生物学功能[J]. 医 学分子生物学杂志, 2009, 6(2): 138-143. Dong fang, Xiao fan, Wei hongshan. Bisecting N-acetylglucosamine and its biological functions[J]. Journal of Medical Molecular Biology, 2009, 6(2): 138-143.