摘要
目的:研究siRNA c-Met腺病毒转染对培养的人外泌汗腺上皮细胞(human eccrine sweat gland epithelial cells,hESGc)增殖的影响。方法:按我科已经建立的方法培养和鉴定hESGc,取第1代细胞备用。构建表达红色荧光蛋白的siRNA c-Met腺病毒,荧光显微镜观察和Westernlot检测证实病毒的有效性。MTT法检测siRNA c-Met腺病毒对hESGc增殖的影响。结果:荧光显微镜下可见转染了siRNA c-Met腺病毒的hESGc有红色荧光表达,转染后48h,收集细胞进行westernblot检测,结果发现,siRNA c-Met腺病毒转染能抑制hESGc细胞中c-Met蛋白的表达,抑制率达70%以上,MTT法检测发现转染siRNA c-Met腺病毒能抑制hESGc增殖,抑制率分别为16.8%(2天)和34.5%(4天)。结论:siRNA c-Met腺病毒转染能抑制hESGc的增殖。
Objective To investigate the effect of siRNA c-Met adenovirus on proliferation of cultured hESGc. Methods The recombinant adenoviral vector containing siRNA targeting c-Met gene was successfully constructed. Adenovirus with high titer was harvested. Fluorescence microscope observation and westernblot test were used to confirm the effectiveness of adenovirus. MTT assay was used to detect the growth of the cells. Results Red fluorescence was observed in hESGc infected with siRNA c -Met adenovirus under fluorescence microscopy. The infected cells were harvested for westernblot test. It is observed that c-Met was inhibited in siRNA c-Met adenovirus infected cell group. The transfection of siRNA c-Met adenovirus can inhibit the growth of hESGc by MTT assay. The inhibition rate were 16.8% in 2 days and 34.5% in 4 days. Conclusion The transfection of siRNA c-Met adenovirus can inhibit the growth of hESGc.
出处
《中国美容医学》
CAS
2010年第9期1324-1326,共3页
Chinese Journal of Aesthetic Medicine
基金
国家自然科学基金资助项目(30801193
30700793)
第三军医大学第三附属医院"1135"创新人才基金(504-1371)
关键词
C-MET
外泌汗腺
上皮细胞
增殖
c-Met
eccrine sweat gland
epithelial cell
proliferation
