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
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摘要
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.
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.
引文
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[18]Mo C,Liu T,Zhang S,et al.Reduced N-acetylglucosaminyltransferase III expression via Smad3 and Erk signaling in TGF-β1-induced HCC EMT model[J].Discov Med,2017,23(124):7-17.(收稿日期:2017-10-27)
[2]Ma H,Zhou H,Li P,et al.Effect of ST3GAL4 and FUT7 on sialyl Lewis X synthesis and multidrug resistance in human acute myeloid leukemia[J].Biochim Biophys Acta,2014,1842(9):1681-1692.
[3]Ogawa J I,Inoue H,Koide S.alpha-2,3-Sialytransferase type3N and alpha-1,3-fucosyltransferase typeⅦare related to sialyl Lewis(x)synthesis and patient survival from lung carcinoma[J].Cancer,1997,79(9):1678-1685.
[4]Kannagi R,Izawa M,Koike T,et al.Carbohydrate-mediated cell adhesion in cancer metastasis and angiogenesis[J].Cancer Sci,2004,95(5):377-384.
[5]Siegel R L,Miller K D,Jemal A.Cancer statistics,2017[J].CACancer J Clin,2017,67(1):7-30.
[6]Reimand J,Wagih O,Bader G D.Evolutionary constraint and disease associations of post-translational modification sites in human genomes[J].PLoS Genet,2015,11(1):e1004919.
[7]Lis H,Sharon N.Protein glycosylation.Structural and functional aspects[J].Eur J Biochem,1993,218(1):1-27.
[8]Pinho S S,Reis C A.Glycosylation in cancer:mechanisms and clinical implications[J].Nat Rev Cancer,2015,15:540-555.
[9]Becker D J,Lowe J B.Fucose:biosynthesis and biological function in mammals[J].Glycobiology,2003,13(7):41R-53R.
[10]Li W,Zhang W,Luo J,et al.Alpha1,3 fucosyltransferaseⅦplays a role in colorectal carcinoma metastases by promoting the carbohydration of glycoprotein CD24[J].Oncol Rep,2010,23(6):1609-1617.
[11]Wang H,Wang Q Y,Zhang Y,et al.Alpha1,3 Fucosyltransferase-Ⅶmodifies the susceptibility of apoptosis induced by ultraviolet and retinoic acid in human hepatocarcinoma cells[J].Glycoconj J,2007,24(4-5):207-20.
[12]Seales E C,Jurado G A,Brunson B A,et al.Hypersialylation of beta1 integrins,observed in colon adenocarcinoma,may contribute to cancer progression by up-regulating cell motility[J].Cancer Res,2005,65(11):4645-4652.
[13]Komatsu H,Mizuguchi S,Izumi N,et al.Sialyl lewis X as a predictor of skip N2 metastasis in clinical stage IA non-small cell lung cancer[J].World J Surg Oncol,2013,11:309.
[14]Liang J X,Liang Y,Gao W.Clinicopathological and prognostic significance of sialyl Lewis X overexpression in patients with cancer:a meta-analysis[J].Onco Targets Ther,2016,9:3113-3125.
[15]Tei K,Kawakami-Kimura N.Roles of cell adhesion molecules in tumor angiogenesis induced by cotransplantation of cancer and endothelial cells to nude rats[J].Cancer Res,2002,62(21):6289-6296.
[16]Yilmaz M,Christofori G.EMT,the cytoskeleton,and cancer cell invasion[J].Cancer Metastasis Rev,2009,28(1-2):15-33.
[17]Wang A,Lu C,Ning Z,et al.Tumor-associated macrophages promote Ezrin phosphorylation-mediated epithelial-mesenchymal transition in lung adenocarcinoma through?FUT4/LeYup-regulation[J].Oncotarget,2017,8(17):28247-28259.
[18]Mo C,Liu T,Zhang S,et al.Reduced N-acetylglucosaminyltransferase III expression via Smad3 and Erk signaling in TGF-β1-induced HCC EMT model[J].Discov Med,2017,23(124):7-17.(收稿日期:2017-10-27)