摘要
目的:通过腺病毒载体介导的RNA干扰技术抑制乳腺癌细胞HGF受体c-Met的表达,抑制乳腺癌细胞增殖、诱导细胞凋亡。方法:PCR法获得人U6启动子及带有c-Met反向互补靶序列的片段HU6shmet;利用腺病毒载体将其传递至SK-BR-3细胞;RNA狭缝杂交检测SK-BR-3细胞c-Met mRNA水平,Western印迹检测c-Met蛋白水平。结果:获得了带有人U6启动子及c-Met反向互补靶序列的重组腺病毒载体rAdUshmet1和rAdUshmet2。转导了重组腺病毒的SK-BR-3细胞的c-Met mRNA和蛋白相对表达量均有所下降。结论:腺病毒载体rAdUshmet通过抑制HGF受体c-Met表达,能在一定程度上阻断HGF-c-Met信号转导通路,有可能成为对乳腺癌进行基因治疗的有效载体。
Objective： To inhibit the expression of hepatocyte growth factor （HGF） receptor-c-Met by RNAi technique mediated by adenovirus vector, so as to inhibit tumor cell proliferation and induce tumor cell apoptosis. Methods： Human U6 promotor and the fragment including c-Met reverse complement target sequence were obtained by PCR. The fragment was packaged in adenovirus and transduced into SK-BR-3 cells, c-Met mRNA level was assayed by slit hybridization and the protein level was assayed by Western blot. Results： The recombinated adenovirus vector （named rAdUshmetl and rAdUshmet2）, harboring human U6 promotor and c-Met reverse complement target sequence, were obtained. The c-Met mRNA and protein expression levels were reduced in the SK-BR-3 cells transducted with the rAdUshmetl and rAdUshmet2. Conclusion： The c-Met short hairpin RNA mediated by adenovirus vector can inhibit c-Met expression, block the HGF-c-Met signal pathway partially and may act as an effective vector for gene therapy of breast cancer.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2006年第5期549-552,共4页
Chinese Journal of Anatomy
