过表达乙酰肝素酶促进胆囊癌细胞系GBC-SD的增殖和迁移

目的探讨乙酰肝素酶基因过表达对胆囊癌细胞系(GBC-SD)生物学功能的影响。方法构建乙酰肝素酶过表达质粒载体,通过脂质体法转染至胆囊癌细胞系GBC-SD中。用RT-qPCR和Western blot验证转染效果。CCK-8法、流式细胞计量术、划痕愈合实验检测细胞增殖、凋亡、迁移。Western blot检测syndecan-1、FGF-2、E-cadherin蛋白表达情况。结果 GBC-SD细胞经转染后乙酰肝素酶mRNA(P<0. 01)和蛋白(P<0. 05)表达水平显著升高。与对照组相比,过表达乙酰肝素酶显著上调GBC-SD细胞体外增殖和迁移能力(P<0. 05),下调凋亡水平(P<0. 01),FGF-2蛋白表达显著升高(P<0. 05),而syndecan-1(P<0. 01)和E-cadherin(P<0. 05)蛋白表达降低。结论乙酰肝素酶过表达促进胆囊癌细胞系GBC-SD的增殖与迁移,其机制可能与FGF-2调节E-cadherin有关。

Over-expression of heparanase promotes proliferation and migration of gallbladder cancer cell line GBC-SD

Objective To investigate the effect of heparanase over-expression on the biological function of gallbladder carcinoma cell line GBC-SD.Methods The heparanase over-expression plasmid vector was constructed and transfected into GBC-SD cells by liposome method.The transfection efficiency was verified by RT-qPCR and Western blot.CCK-8 assay,flow cytometry and scratch healing experiment were used to detect the cell proliferation,apoptosis and migration.The expression of syndecan-1,FGF-2 and E-cadherin protein was detected by Western blot.Results The expression of heparanase mRNA(P<0.01)and protein(P<0.05)in gallbladder cancer cell line GBC-SD was significantly increased after transfection.Compared with the control group,the over-expression of heparanase significantly up-regulated the proliferation and migration ability of GBC-SD cells in vitro(P<0.05),down-regulated the level of apoptosis(P<0.05),and increased the expression of FGF-2 protein,while decreasing the expression of syndecan-1(P<0.01)and E-cadherin(P<0.05).Conclusions Over-expression of heparanase promotes the proliferation and migration of gallbladder cancer cell line GBC-SD,and the mechanism may be related to its regulation of E-cadherin through FGF-2.