Cell surface glycan alterations in epithelial mesenchymal transition process of Huh7 hepatocellular carcinoma cell.
Cell surface glycan alterations in epithelial mesenchymal transition process of Huh7 hepatocellular carcinoma cell.
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Huh7肝癌细胞上皮间质转化过程中细胞表面聚糖的改变
DOI：
10.1371/journal.pone.0071273
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Liu Y
中科院分区：
文献类型：
--
作者：
Li S;Mo C;Peng Q;Kang X;Sun C;Jiang K;Huang L;Lu Y;Sui J;Qin X;Liu Y
背景与目的肝细胞癌（Hepatocellular carcinoma，HCC）由于复发和转移，死亡率高.众所周知，上皮间质转化（EMT）和细胞表面糖蛋白聚糖在肿瘤转移中起着关键作用。本研究的目的是使用HGF诱导的EMT模型鉴定HCC转移相关的差异聚糖模式及其酶学基础。方法采用肝细胞生长因子诱导肝癌EMT模型。用凝集素芯片检测细胞表面糖蛋白的表达，并通过凝集素印迹和荧光细胞凝集素免疫化学方法验证差异。通过qRT-PCR测定糖基转移酶的mRNA表达水平。结果经HGF处理后，Huh 7细胞失去上皮性特征，获得间充质标志。这些变化表明HGF可诱导典型的EMT细胞模型。凝集素微阵列分析鉴定了7种凝集素ACL、BPL、JAC、MPL、PHA-E、SNA和SBA对细胞表面糖蛋白聚糖的亲和力降低。这意味着含有T/Tn抗原、NA 2和二等分GlcNAc、Siaα2- 6 Gal/GalNAc、末端α或βGalNAc结构的聚糖减少。AAL、LCA、LTL、ConA、NML、NPL、DBA、HAL、PTL II、WFL、ECL、GSL II和PHA-L等13种凝集素与糖链的结合能力增强，并明确表明含有末端αFuc和± Sia-Le、核心岩藻糖、α-man、gal-β（α）GalNAc、β 1，6 GlcNAc分支和四触角复合寡糖结构的糖链增加。这些结果进一步验证了凝集素印迹和荧光细胞凝集素免疫化学。此外，Mgat 3的mRNA表达水平降低，而Mgat 5、FucT 8和β 3GalT 5的mRNA表达水平升高。因此，EMT过程中细胞表面聚糖的改变可能与糖基转移酶的表达一致。结论本研究系统阐明了肝癌EMT中细胞表面糖链的变化，为肝癌转移提供了新的认识。
Background and Objective Due to recurrence and metastasis, the mortality of Hepatocellular carcinoma (HCC) is high. It is well known that the epithelial mesenchymal transition (EMT) and glycan of cell surface glycoproteins play pivotal roles in tumor metastasis. The goal of this study was to identify HCC metastasis related differential glycan pattern and their enzymatic basis using a HGF induced EMT model. Methodology HGF was used to induce HCC EMT model. Lectin microarray was used to detect the expression of cell surface glycan and the difference was validated by lectin blot and fluorescence cell lectin-immunochemistry. The mRNA expression levels of glycotransferases were determined by qRT-PCR. Results After HGF treatment, the Huh7 cell lost epithelial characteristics and obtained mesenchymal markers. These changes demonstrated that HGF could induce a typical cell model of EMT. Lectin microarray analysis identified a decreased affinity in seven lectins ACL, BPL, JAC, MPL, PHA-E, SNA, and SBA to the glycan of cell surface glycoproteins. This implied that glycan containing T/Tn-antigen, NA2 and bisecting GlcNAc, Siaα2-6Gal/GalNAc, terminal α or βGalNAc structures were reduced. The binding ability of thirteen lectins, AAL, LCA, LTL, ConA, NML, NPL, DBA, HAL, PTL II, WFL, ECL, GSL II and PHA-L to glycan were elevated, and a definite indication that glycan containing terminal αFuc and ± Sia-Le, core fucose, α-man, gal-β(α) GalNAc, β1,6 GlcNAc branching and tetraantennary complex oligosaccharides structures were increased. These results were further validated by lectin blot and fluorescence cell lectin-immunochemistry. Furthermore, the mRNA expression level of Mgat3 decreased while that of Mgat5, FucT8 and β3GalT5 increased. Therefore, cell surface glycan alterations in the EMT process may coincide with the expression of glycosyltransferase. Conclusions The findings of this study systematically clarify the alterations of cell surface glycan in cancer EMT, and may provide novel insight for HCC metastasis.
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DOI：
10.1016/j.biocel.2007.07.011
发表时间：
2007-01-01
影响因子：
4
作者：
Guarino, Marcello
通讯作者：
Guarino, Marcello
影响因子：
3.7
作者：
Comunale MA;Rodemich-Betesh L;Hafner J;Wang M;Norton P;Di Bisceglie AM;Block T;Mehta A
通讯作者：
Mehta A
影响因子：
3.7
作者：
Pinho SS;Oliveira P;Cabral J;Carvalho S;Huntsman D;Gärtner F;Seruca R;Reis CA;Oliveira C
通讯作者：
Oliveira C
影响因子：
3.4
作者：
Dai, Zhi;Liu, Yin-Kun;Tang, Zhao-You
通讯作者：
Tang, Zhao-You
影响因子：
254.7
作者：
Parkin, DM;Bray, F;Pisani, P
通讯作者：
Pisani, P