靶向STAT5的siRNA诱导人肝癌SMMC-7721细胞凋亡  

siRNA targeting STAT5 induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721

作　　者：赵振军 张丽杰[2] 鹿刚 张引娟[3] 单保恩[3] 

机构地区：[1]河北省以岭医院检验科,石家庄050091 [2]河北医科大学第三医院检验科,石家庄050051 [3]河北医科大学第四医院科研中心,石家庄050011

出　　处：《第三军医大学学报》2009年第12期1147-1150,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(3037153);河北省自然科学基金(C2004000610)~~

摘　　要：目的采用RNA干扰技术阻断STAT5基因的表达,观察其对人肝癌SMMC-7721凋亡的影响。方法构建3种靶向STAT5的小干扰RNA(siRNA)真核表达载体,采用LipofectamineTM2000转染肝癌细胞SMMC-7721;分别采用半定量RT-PCR、Western blot技术检测转染前后SMMC-7721细胞STAT5 mRNA和蛋白质表达的变化;采用透射电镜技术观察细胞形态变化;流式细胞术检测细胞凋亡率。结果3条靶向STAT5的序列特异性的siRNA可以有效地抑制SMMC-7721细胞STAT5 mRNA和蛋白质表达,STAT5 mRNA表达抑制率分别为70.43%、43.02%、45.07%,STAT5蛋白表达抑制率分别为67.45%、37.36%、41.86%;转染靶向STAT5的siRNA真核表达载体48h后出现凋亡细胞的典型形态学变化,并诱导25.61%的SMMC-7721细胞发生凋亡。结论靶向STAT5的siRNA真核表达载体可以有效、特异地阻断SMMC-7721细胞STAT5基因的表达,并能够诱导SMMC-7721细胞凋亡,STAT5 siRNA在人肝癌的基因治疗中可能具有潜在的应用价值。Objective To investigate the effects of inhibition of STATS gene expression by RNA interference technology on apoptosis of human hepatocellular carcinoma cell line SMMC-7721. Methods Three siRNA eukaryotic expression vectors against STATS were constructed and transfected with lipofectamineTM 2000 into SMMC-7721 cells. The changes in STATS expression were detected by semi-quantitative RT-PCR and Western blot. Cell apoptosis was assayed by flow cytometry （FCM）. Results The sequence-specific siRNA could effectively and specifically inhibit STATS gene expression at both mRNA and protein levels. The inhibition rates of STATS mRNA expression were 70.43%, 43.02% , and 45.07% , respectively. The inhibition rates of STATS protein expression were 67.45%, 37.36%, and 41.86%, respectively. At 48 h after transfection, apoptosis rate was 25.61%. Conclusion siRNA against STATS can inhibit STATS gene expression in SMMC-7721 cells effectively and specifically and induce apoptosis of SMMC-7721 cells, siRNA targeting STATS has a great potential value in gene therapy of hepatocellular carcinoma.

关 键 词：STAT5 SIRNA 凋亡 人肝癌细胞SMMC-7721 

分 类 号：R73-362[医药卫生—肿瘤] R735.7[医药卫生—临床医学]