HDGF过表达促进胶质瘤细胞体外迁移与侵袭及β-catenin磷酸化

HDGF overexpression enhances migration and invasion of glioma cells by promoting β-catenin phosphorylation in vitro

目的:研究上调肝癌衍生生长因子(hepatoma-derived growth factor,HDGF)表达对胶质瘤细胞体外生物学行为的影响。方法:通过MTS实验和溴代脱氧尿苷(bromodeoxyuridine,BrdU)掺入实验,研究过表达HDGF对恶性胶质瘤细胞株DBTRG体外增殖的影响。采用划痕实验、Transwell小室迁移实验和侵袭实验,研究过表达HDGF对DBTRG细胞体外迁移与侵袭能力的影响。通过Western blot和免疫荧光实验研究上调HDGF表达对细胞黏附分子的表达及亚细胞定位的影响。结果:过表达组与对照组细胞在增殖率和新生细胞比例上均无明显差异。迁移和侵袭实验结果显示,HDGF过表达细胞较对照细胞的迁移距离、细胞数量及侵袭细胞数均显著增加。此外,HDGF过表达可增加N-cadherin表达但降低β-catenin蛋白水平。进一步的结果显示HDGF过表达不影响β-catenin核转位,但可促进其磷酸化。结论:上调HDGF表达可显著增强DBTRG细胞体外迁移与侵袭能力,这一作用可能与HDGF促进β-catenin磷酸化有关。

Objective:To investigate the effects of up-regulation of hepatoma-derived growth factor(HDGF)expression on the biological behavior of glioma cells in vitro. Methods: The effects of overexpression of HDGF on the proliferation of malignant glioma cell line DBTRG were examined by MTS and bromodeoxyuridine(BrdU)incorporation experiments. The effects of HDGF up-regulation on the migration and invasion of DBTRG in vitro were investigated by Wound-healing assay,Transwell assay and Matrigel invasion assay,respectively. Western blot and immunofluorescence were used to determine the expression and subcellular localization of cell adhesion molecules. Results: There was no significant difference in the proliferation rate and the proportion of BrdU positive cells between the overexpressing group and the control group. Compared with the control group,the numbers of migration and invasion cells in the HDGF overexpression group were significantly increased. In addition,overexpression of HDGF increased N-cadherin expression but decreased β-catenin protein levels. Furthermore,HDGF did not affect the nuclear translocation of β-catenin,but promoted its phosphorylation. Conclusion: HDGF enhances migration and invasion of DBTRG glioma cells in vitro by promoting β- catenin phosphorylation.