与肿瘤侵袭相关的Lewis抗原在人肝癌细胞H7721上的表达及其合成的酶学分析

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目的 研究人肝癌细胞H772 1表面 4种含岩藻糖类抗原—Lewis抗原的表达、它们与肝癌细胞在体外侵袭行为的关系 ,及其合成相关酶α1,3岩藻糖转移酶 (FucT) 5种亚型的基因表达。方法 用单克隆抗体结合流式细胞仪测定Lewis抗原 ,用涂有Matrigel膜的转移小室测定细胞的侵袭行为 ,利用实时RT PCR测定FucT的表达。结果 H772 1细胞表面主要表达Slex和少量SDLex,而Lex和Slea表达极微。其中只有Slex和H772 1细胞的侵袭明显相关。H772 1细胞中FucT的表达强度为FucT IV >FucT III >FucT VI≥FucT VII,后两者表达少 ,而几乎完全缺乏FucT IX。结论 Slex是H772 1细胞表面最多也是与细胞侵袭最有关的Lewis抗原 ,因据报道FucT VI催化合成Slex和SDLex的效率高于FucT III及VII,故可能是负责H772 1细胞中Slex和SDLex合成的主要FucT ,但也不排除FucT VII和FucT III也参与Slex的合成。合成Lex中最重要的FucT IX的缺乏可能是Lex低表达的原因。 Objective To study the expression of four anti-Lewis antigens containing fucosylated antigens on the surface of human hepatocellular carcinoma cell line H772 1 and their relationship with the invasiveness of human hepatocellular carcinoma cells in vitro and to explore the mechanisms underlying their biosynthesis-related enzymes α1,3 fucosyltransferase (FucT) 5 Subtype of gene expression. Methods The Lewis antigen was detected by monoclonal antibody combined with flow cytometry. The invasion of cells was measured by the transfer chamber coated with Matrigel membrane, and the expression of FucT was detected by real-time RT-PCR. Results The cell surface of H772 1 mainly expressed Slex and a small amount of SDLex, while the expressions of Lex and Slea were minimal. Among them, only the invasion of Slex and H772 1 cells was significantly correlated. The intensity of FucT expression in H772 1 cells was FucT IV> FucT III> FucT VI ≧ FucT VII, with little expression of the latter two and almost complete lack of FucT IX. Conclusions Slex is the most antigens involved in cell invasion on H772 1 cells and Lewis ligand. It has been reported that FucT VI catalyzes the synthesis of Slex and SDLex more efficiently than FucT III and VII, so it may be responsible for the synthesis of Slex and SDLex in H772 1 cells The main FucT, but does not rule out FucT VII and FucT III also involved in the synthesis of Slex. The lack of the most important FucT IX in the synthesis of Lex may be responsible for the low expression of Lex.
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