eIF-4E腺病毒载体构建及对乳腺癌MCF-7细胞转移能力的影响  

Construction of eIF-4E Adenovirus and Its Effect on Metastasis Ability of Human Breast Cancer Cell Line MCF-7

作　　者：张国强[1] 潘薇[1] 赵亮[1] 庞达[1] 

机构地区：[1]哈尔滨医科大学附属第三医院,哈尔滨150081

出　　处：《中国生物工程杂志》2008年第3期8-12,共5页China Biotechnology

基　　金：黑龙江省青年基金资助项目(QC05C48)

摘　　要：目的:构建乳腺癌真核细胞起始因子——4E(eukaryotic initiation factor 4E,eIF-4E)基因重组腺病毒载体,并观察其对乳腺癌细胞MCF-7转移能力的影响。方法:应用基因重组技术将eIF-4E基因构建于腺病毒载体pAD-X,转染293包装细胞得到高滴度重组腺病毒,并用real-time PCR进行eIF-4E基因表达的验证。将重组腺病毒pAD-eIF-4E感染MCF-7细胞,应用siRNA封闭eIF-4E后通过transwell小室法观察其对细胞侵袭和运动能力的影响。结果:酶切结果与预期相符,real-time PCR可检测到转染后MCF-7细胞有eIF-4E基因过表达。且病毒转染后transwell小室可见,与eIF-4E过表达组相比较,eIF-4E siRNA封闭组MCF-7细胞的侵袭和运动能力均受到显著的抑制(均为p<0.01)。结论:重组eIF-4E腺病毒载体正确构建,抑制eIF-4E的表达,对乳腺癌细胞MCF-7的侵袭和运动都有抑制作用。Objective： To reconstruct adenovirus vector of breast eukaryotic initiation factor 4E and to observe its effect on the metastasis ability of breast cancer cell line MCF-7. Methods： eIF-4E gene was constructed into adenovirus vector pAD-X by gene recombination technique, which was transformed into 293 packaged cell for high titer adenovirus. Real-time PCR was applied to detect eIF-4E gene expression, eIF 4E siRNA was applied and then transwell cabin assay was used to observe changes of invasion and motor ability of MCF-7 ceils transfected with reconstruction adenovirus. Result： The finding of digestion was coincided with expected, eIF-4E gene over-expression was detected in transfected MCF-7 cells with real-time PCR. And the invasion and motor abilities of transfected MCF-7 cells were more significantly inhibited in transwell cabin assay （respectively p 〈0.01 ） in eIF-4E siRNA group than that in controls. Conclusion： eIF-4E adenovirus was correctly reconstructed and invasion and motor abilities of MCF-7 ceils could be inhibited while eIF-4E expression was silent.

关 键 词：EIF-4E 重组腺病毒 乳腺癌MCF-7细胞 RNA干扰 侵袭和运动 

分 类 号：Q789[生物学—分子生物学]