STAT5A基因有效siRNA序列的筛选及其对人肝癌HepG2细胞增殖及凋亡的影响

目的筛选针对STAT5A基因有效的siRNA，研究抑制STAT5A基因的表达对人肝癌HepG2细胞增殖、周期及凋亡的影响，探讨STAT5A基因在肝癌发生发展中的作用。方法设计并化学合成针对STAT5A基因三个靶点的siRNA，采用脂质体法转染人肝癌细胞HepG2，分别用半定量RT-PCR、Westernblot技术检测转染后HepG2细胞STAT5AmRNA和蛋白表达的变化，从中筛选出一段干扰效率最显著的siRNA；用该siRNA转染HepG2细胞，MTT法检测细胞增殖情况，流式细胞仪检测细胞凋亡率。结果三段STAT5A基因的siRNA均对HepG2细胞中STAT5AmRNA和蛋白表达产生了不...

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Published in肿瘤防治研究 Vol. 40; no. 11; pp. 1031 - 1035
Main Author 杨华秀 曾永秋 曹洋 林春燕 黄燕 李洁
Format Journal Article
LanguageChinese
Published 2013
Subjects
HepG2细胞
siRNA
STAT5A基因
细胞凋亡
细胞增殖
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ISSN1000-8578

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Summary:目的筛选针对STAT5A基因有效的siRNA，研究抑制STAT5A基因的表达对人肝癌HepG2细胞增殖、周期及凋亡的影响，探讨STAT5A基因在肝癌发生发展中的作用。方法设计并化学合成针对STAT5A基因三个靶点的siRNA，采用脂质体法转染人肝癌细胞HepG2，分别用半定量RT-PCR、Westernblot技术检测转染后HepG2细胞STAT5AmRNA和蛋白表达的变化，从中筛选出一段干扰效率最显著的siRNA；用该siRNA转染HepG2细胞，MTT法检测细胞增殖情况，流式细胞仪检测细胞凋亡率。结果三段STAT5A基因的siRNA均对HepG2细胞中STAT5AmRNA和蛋白表达产生了不同程度的抑制作用，其中以siRNA-3697的干扰效率最高，STAT5AmRNA和蛋白的表达抑制率分别为72．03％和66．27％（P〈0．05）；转染后1-4d细胞生长速度减慢、增殖显著抑制（P〈0．05）；转染后48h细胞凋亡率为37．33％（P〈0．05）。结论成功筛选出针对STAT5A基因有效的siRNA；该siRNA特异地阻断HeDG2细胞STAT5A基因的表达．抑制细胞生长，诱导细胞凋亡。
Bibliography:STAT5A gene; siRNA; HepG2 cell; Cell proliferation; Cell apoptosis
42-1241/R
YANG Huaxiu , ZENG Yongqiu , CAO Yang LIN Chunyan HUANG Yan, LI Jie （ 1. Department of Hepatopathy, Affiliated Hospital（T.C.M） Affiliated to Luzhou Medical College, Luzhou 646000, China;2.Department of Medical Biology and Genetics, Luzhou Medical College; 3. Department of Physiology, Luzhou Medical College, Luzhou）
Objective To screen for effective siRNAs for STAT5A gene and study the effect of suppression of STATSA gene expression on proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2, and to explore the role of STAT5A in generation and development of hepatocellular carcinoma. Methods Three siRNAs targeted at STAT5A gene were designed and synthesized chemically, and then transfected into HepG2 cells with liposome transfection method. The expression levels of STATSA mRNA and protein were detected by semi-quantitative reverse transcript polymerase chain reaction （RT-PCR） and immunoblotting assay （Western blot） r
ISSN:1000-8578