摘要
目的探讨HER4在食管癌细胞侵袭和转移过程中的作用。方法应用RNA干扰技术抑制HER4在食管癌细胞系Eca-109中的表达,Western blot方法观察细胞MMP-9、VEGF蛋白表达水平的变化,细胞划痕实验和Transwell侵袭实验观察细胞迁移、侵袭能力的变化。结果酶切鉴定和DNA测序分析显示HER4靶向RNA干扰重组载体构建成功;荧光实时定量PCR和Westernblot检测筛选得到最佳干扰效果质粒;该质粒转染细胞后,MMP-9、VEGF蛋白表达水平出现明显降低,细胞迁移、侵袭能力显著下降。结论HER4与食管癌的侵袭转移潜能相关,敲减HER4的表达可明显抑制食管癌细胞系Eca-109的运动和侵袭能力。
Objective To investigate the role of HER4 in cell invasion and metastasis of human esophageal carcinoma. Methods Knockdown of HER4 was done by stably expressing short hairpin RNA （shRNA） specific for HER4 mRNA in a malignant esophageal carcinoma cell line Eea-109. The expressions of MMP-9 and VEGF proteins were detected by Western blot assay after RNA interference. Wound-healing scrape assay and transwell invasion assay were performed to assess the effect of HER4-targeted RNA interference on the invasive properties of esophageal carcinoma in vitro. Results Enzyme digestion analysis and DNA sequencing showed that HER4-targeted RNA interference plasmid was constructed successfully. The most effective vector plasmid was selected by Real-time quantitative PCR and Western blot assay. After transfection, down-regulation of MMP-9 and VEGF expression was observed, and knockdown of HER4 significantly impaired the migration and invasion of Eca-109 cells in vitro. Conclusions HER4 is correlated with the invasive and metastatic potential of esophageal carcinoma, and knockdown of HER4 induced the inhibition of migration and invasion of Eea-109 cell line.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2008年第4期521-527,共7页
Fudan University Journal of Medical Sciences
