RNA干扰对肝癌细胞株SMMC7721中STAT3基因表达的抑制作用  

The inhibitory effect of RNA interference on STAT3 expression in liver cancer cell line SMMC7721

作　　者：廖维甲[1,2] 梅铭惠[1,2] 覃理灵[1,2] 陈谦[1,2] 袁晟光[1,2] 刘杰[1,2] 

机构地区：[1]桂林医学院附属医院肝胆外科研究室 [2]桂林医学院肝病防治研究所,广西桂林541001

出　　处：《细胞与分子免疫学杂志》2011年第6期615-617,620,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30860324);广西自然科学基金项目资助(桂科自0832268);广西壮族自治区卫生厅重点科研课题资助(重200714;重200883)

摘　　要：目的:研究小片段干扰RNA(siRNA)对肝癌细胞系SMMC7721的化疗敏感性。方法:根据信号转录及转录活化因子3(STAT3)基因设计siRNA序列,通过经脂质体Lipo-fectamineTM2000以siRNA转染SMMC7721细胞。用实时定量PCR检测SMMC7721细胞中STAT3基因表达的抑制。将细胞以10μmol/L5-氟脲嘧啶(5-FU)作用后,用MTT比色法检测细胞生长的抑制率。结果:成功地构建针对STAT3基因的siRNA表达载体。实时定量PCR检测结果显示,SMMC7721细胞经特异性siRNA转染后,STAT3基因的表达受到抑制。RNA干扰(RNAi)能特异、有效地抑制SMMC7721细胞中STAT3基因的表达。MTT比色法检测结果显示,经siRNA作用后SMMC7721细胞抑制率明显增加。结论:设计合成的siRNA表达载体能有效抑制STAT3基因在肝癌细胞系SMMC7721中表达,增强其对化疗药物5-FU的敏感性,为肿瘤的生物学治疗提供了实验依据。AIM： To investigate the chemosensitivity small interfering RNA（siRNA） on liver cancer cell line SMMC7721.METHODS： The siRNA sequences design based on signal transducers and activators of transcription 3（STAT3） gene,siRNA were transfected into SMMC7721 cells through liposome lipofectamineTM 2000.The expression inhibition of STAT3 gene in SMMC7721 cells was measured by real-time relative quantitative PCR.The cells growth inhibition rate were measured by MTT colorimetry after 10 μmol/L 5-fluorouracil（5-Fu） action.RESULTS： The siRNA expression vector to aim directly at STAT3 gene was constructed successfully.The result of real-time PCR revealed that specificity siRNA were transfected into SMMC7721 cells could inhibit the expression of STAT3 gene.STAT3 gene in SMMC7721 cells were specialty and effectually inhibit the expression by RNA interference（RNAi）.MTT colorimetry detection result revealed that SMMC7721 cells inhibition rate significantly increasing after siRNA action.CONCLUSION： The siRNA expression vector can active inhibit expression of STAT3 gene in SMMC7721 cells,enhance its sensitivity to chemotherapeutics 5-Fu,to provide experiment based on for the biological therapy of tumor.

关 键 词：小干扰RNA STAT3 SMMC7721细胞 5-氟脲嘧啶 化疗敏感性 

分 类 号：R735.7[医药卫生—肿瘤]