摘要
目的:建立Grim-19基因稳定表达的人子宫颈癌Hela细胞株,探讨Grim-19及其对下游基因STAT3表达的影响。方法:将构建好的质粒采用脂质体法转染人子宫颈癌Hela细胞株,经G418筛选,得到稳定表达Grim-19的Hela细胞株,并分别通过逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western-blot)检测Grim-19及其下游基因STAT3的表达。结果:成功建立了高表达GRIM-19的Hela/Grim-19细胞株,并在mRNA与蛋白水平上均发现Grim-19在转染细胞中的高表达和其下游基因STAT3表达的明显下调。结论:上调子宫颈癌Hela细胞Grim-19的表达对STAT3的表达有抑制作用。
Objective： To establish the stable Hela cell lines with a persistent expression of GRIM - 19, explore the expression of GRIM- 19 and it＇s effect on the downstream gene -signal transducer and activator of transcription 3 （ STAT3 ） .Methods： The recombinant plasmid was transfected into Hela cells by liposome method, then the positive transfected cells were screened by G418. The expression of GRIM - 19 and STAT3 in Hela/GRIM - 19 cells in transfected Hela cells was identified by RT - PCR and Western blot. Results： The transfencted Hela cell line was established suscessfully, which can express GRIM - 19 highly. The high expression of GRIM - 19 in Hela cells can obviously downregulate the expression of STAT3. Conclusion： The high expression of GRIM - 19 in Hela cells can inhibit the expression of STAT3.
出处
《中国妇幼保健》
CAS
北大核心
2009年第10期1406-1409,共4页
Maternal and Child Health Care of China
基金
安徽省科技攻关项目(No.06013124B)
关键词
GRIM-19
STAT3
转染
HELA细胞
GRIM - 19
Signal transducer and activator of transcription 3
Transfection
Hela cell
