岩藻糖基转移酶Ⅶ通过sLeX糖基化诱导肺癌A549细胞上皮间质转化的研究

Fucosyltransferase Ⅶ induces epithelial-mesenchymal transition of A549 cells via sLeX synthesis

目的研究岩藻糖基转移酶Ⅶ(FUT7)对肺癌A549细胞上皮间质转化(EMT)的影响。方法采用质粒转染法对A549细胞中FUT7基因进行过表达;划痕实验检测细胞侵袭能力;Western blot法检测唾液酸化的路易斯寡糖-X抗原(sLeX)糖抗原以及EMT相关蛋白表达水平。结果转染FUT7过表达质粒后A549细胞中FUT7蛋白的表达明显增强(P<0.01),与空白质粒转染组相比,sLeX抗原表达增高(P<0.05),细胞侵袭能力增强。FUT7过表达质粒转染组与空白质粒转染组相比,E-cadherin蛋白表达减少,N-cadherin、Vimentin蛋白表达均增加(均P<0.01),利用sLeX抗体封闭细胞表面sLeX抗原后FUT7过表达细胞E-cadherin蛋白表达增加(P<0.05),而N-cadherin、Vimentin蛋白表达减少(P<0.05或0.01)。结论 FUT7可通过sLeX糖基化作用诱导肺癌A549细胞发生EMT,导致细胞侵袭转移能力增强。

Objective To investigate the effect of fucosyltransferaseVII(FUT7) gene overexpression on epithelialmesenchymal transition(EMT) in lung cancer A549 cells. Methods A549 cells were transfected with FUT7 cDNA plasmid, and the migration ability was determined by wound healing assay. The expression of sialyl Lewis-X(sLeX) antigen and EMT related proteins were determined by Western Blot. Results The expression of FUT7 protein was significantly augmented in A549 cells after transfected with FUT7 plasmid(P<0.01). Compared with blank plasmid, the expression of sLeX antigen was significantly increased(P<0.05), and the migration ability was enhanced in A549 cells transfected with FUT7 plasmid; the expression of E-cadherin protein was decreased, the expression of N-cadherin and Vimentin protein were increased in A549 cells after transfected with FUT7 plasmid(all P<0.01). While the expression of E-cadherin protein(P<0.05) was increased, the expression of N-cadherin(P<0.05) and Vimentin(P<0.01) were decreased after blockage of sLeX antigen in FUT7 overexpression A549 cells. Conclusion FUT7 overexpression promotes the EMT level and the migration ability of A549 cells via unregulation of sLeX antigen expression.