摘要
目的:探讨缺氧诱导因子1α(HIF-1α)在肝癌上皮-间充质化(EMT)中的作用。方法:采用可调控HIF-1α表达的肝癌HepG2Tet-on-HIF-1α细胞系,首先用real-time PCR与Westernblot方法检测低氧环境中HepG2Tet-on-HIF-1α细胞EMT相关分子(E-cadherin,vimentin,FSP-1)及HIF-1α的mRNA和蛋白表达水平,然后在常氧环境下,采用强力霉素(Dox)诱导HepG2Tet-on-HIF-1α细胞HIF-1α过表达,以及HepG2Tet-on-HIF-1α细胞经Dox处理后再转染HIF-1αsiRNA,观察上述分子的表达情况。结果:低氧处理后,HepG2Tet-on-HIF-1α细胞EMT相关分子及HIF-1α的mRNA和蛋白表达水平较常氧状态下均明显增加(均P<0.05);常氧环境下,Dox能诱导HepG2Tet-on-HIF-1α细胞HIF-1α过表达,同时明显增加EMT相关分子的mRNA和蛋白表达水平(均P<0.05),但转染HIF-1αsiRNA后,Dox的诱导作用被取消。结论:HIF-1α促进HepG2细胞EMT,并可能是肝癌基因治疗的有效靶点。
Objective： To investigate the role of hypoxia inducible factor 1α（HIF-1α） in epithelial-mesenchymal transition（EMT） process of hepatocellular carcinoma. Methods： HIF-1α inducible liver cancer cell line HepG2Tet-on-HIF-1α cells were used.First,the mRNA and protein expressions of EMT-related molecules（E-cadherin,vimentin and FSP-1） and HIF-1α in HepG2Tet-on-HIF-1α cells under hypoxia were determined by real-time PCR and Western blot analysis,respectively.Then,under normoxia condition,the expressions of above molecules were determined in HepG2Tet-on-HIF-1α cells with HIF-1α overexpression induced by doxycycline（Dox）,and HepG2Tet-on-HIF-1α cells transfected with HIF-1α siRNA after Dox treatment. Results： After hypoxia treatment,the mRNA and protein expressions of EMT-related molecules and HIF-1α in HepG2Tet-on-HIF-1α cells were all significantly increased compared with normoxia status（all P〈0.05）.Under normoxia condition,HIF-1α was overexpressed,and the EMT-related molecules expressions were simultaneously increased significantly in HepG2Tet-on-HIF-1α by Dox exposure（all P〈0.05）,but the above effect of Dox was abolished by HIF-1α siRNA transfection. Conclusion： HIF-1α can promote EMT process in HepG2 cells,and probably may be an effective target for liver cancer gene therapy.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2013年第7期885-889,共5页
China Journal of General Surgery
基金
国家自然科学基金资助项目(30371395)
关键词
癌
肝细胞
上皮-间充质化
缺氧诱导因子1
Α亚基
Carcinoma
Hepatocellular
Epithelial-Mesenchymal Transition
Hypoxia-Inducible Factor 1
α Subunit
