目的探讨ECRG2基因在人类纤维肉瘤侵袭和转移过程中的作用。方法应用RNA干扰技术抑制内源性ECRG2基因在人类纤维肉瘤HT1080细胞中的表达,建立ECRG2基因在该细胞中的可诱导表达系统,Westernblotting方法观察细胞ECRG2蛋白表达水平的变化,细胞划痕实验和Transwell侵袭实验观察HT1080细胞迁移、侵袭能力的变化。结果Westernblotting检测筛选得到敲除内源性ECRG2基因的最佳Tet—On可诱导表达克隆。细胞划痕实验和Transwell侵袭实验发现,敲除内源性ECRG2基因明显促进HT1080细胞的侵袭迁移能力,而表达外源ECRG2基因的HT1080细胞侵袭、迁移能力显著下降。结论ECRG2基因与人类纤维肉瘤的侵袭转移潜能相关,外源ECRG2基因的表达可明显抑制人类纤维肉瘤HT1080细胞的侵袭、迁移能力。
Objective To investigate the role of ECRG2 gene in migration and invasion of human fibrosarcoma HT1080 cells. Methods Tet-on system was used to set up ECRG2-inducible cell clones. Knockdown of ECRG2 gene was done by stably expressing short hairpin RNA (shRNA) specific for ECRG2 mRNA in a highly invasive fibrosareoma cell line HT1080. The expressions of ECRG2 proteins were detected by Western blotting assay. Wound-healing scrape assay and transwell invasion assay were performed to assess the effect of ECRG2 gene on the invasive properties of HT1080 in vitro. Results The most effective ECRG2- siRNA interference and inducible was selected by Western blotting assay. Knockdown of ECRG2 gene significantly promoted the migration and invasion of HTI080 cells while expression of ECRG2 impaired the migration and invasion of HT1080 cells in vitro. Conclusion ECRG2 gene is correlated with the invasive and metastatic potential of fibrosarcoma carcinoma, and expression of ECRG2 induces the inhibition of migration and invasion of HT1080 cell line.