GRIM-19基因表达及沉默对宫颈癌细胞株SiHa细胞中核转录因子STAT3表达的影响  

Effects of expression and inhibition of GRIM-19 gene on the expression of the nuclear transcription factor STAT3 in the cervical cancer cell line SiHa

作　　者：朱靖[1] 卫莹[1] 王青元[1] 齐丽敏[1] 李敏[1] 肖卫华[2] 凌斌[1] 周颖[1] 

机构地区：[1]安徽医科大学附属省立医院妇产科分子实验室,合肥230001 [2]中国科技大学生命科学院

出　　处：《现代妇产科进展》2010年第4期245-248,共4页Progress in Obstetrics and Gynecology

基　　金：安徽省自然科学基金项目(No:090413117)

摘　　要：目的:构建pEGFP-C2-GRIM-19真核表达载体并设计GRIM-19siRNA干扰序列,以探讨GRIM-19表达水平的变化对SiHa细胞中核转录因子STAT3表达的影响。方法:用RT-PCR法从SiHa细胞中扩增出带HindⅢ、BamHI酶切位点的GRIM-19基因片段,将GRIM-19基因片段克隆到增强型绿色荧光蛋白(EGFP)基因真核表达载体pEGFP-C2上。分别将pEGFP-C2-GRIM-19及GRIM-19siRNA通过脂质体转染入SiHa细胞48h后,通过Western-blot检测GRIM-19、STAT3的表达。结果:成功构建pEGFP-C2-GRIM-19真核表达载体,转染后可见GFP表达及STAT3表达降低,转染GRIM-19siRNA后可见GRIM-19表达降低及STAT3表达升高。结论:在宫颈癌细胞株中GRIM-19具有调控STAT3表达水平的作用。Objective：To construct the pEGFP-C2-GRIM19 eukaryotic expression vector and design GRIM-19 siRNA disturbance sequence,to approach the effects of the changing expression of GRIM-19 on the expression of STAT3 in SiHa cell.Methods：GRIM-19 gene sequence with HindⅢ and BamHI restriction enzyme cutting site were amplified from total RNA of SiHa cell line by reverse transcription-polymerase chain reaction（RT-PCR）,then the GRIM-19 gene sequence was inserted into the eukaryotic expression vector pEGFP-C2 encoding the enhanced green fluorescent protein（EGFP） gene.The plasmid pEGFP-C2-GRIM-19 and GRIM-19 siRNA were transfected into the SiHa cells.Western-blot was used to test the expression levels of GRIM-19 and STAT3 after 48 hours.Results：The pEGFP-C2-GRIM-19 eukaryotic expression vector was successfully constructed.Consistent expression of GFP and decreased expression of STAT3 were seen after transfection of pEGFP-C2-GRIM-19,however,low expression of GRIM-19 and overexpression of STAT3 after transfection of the GRIM-19 siRNA.Conclusion：GRIM-19 can regulating the expression level of STAT3 in the cervical cancer cell line.

关 键 词：宫颈肿瘤 SIHA细胞 基因 GRIM-19 核转录因子STAT3 转染 

分 类 号：R737.33[医药卫生—肿瘤]