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siRNA沉默胃癌SGC-7901细胞HO1基因的细胞系的建立及其生物学行为 被引量：1

Construction and functional analysis of siRNA expression vector of HO1 gene in gastric cancer SGC-7901 cells
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摘要 目的构建HO1基因的真核干扰表达载体,评估其转染人胃癌细胞系SGC-7901细胞后对HO1基因的干扰效果及其功能。方法将外源性重组真核干扰表达载体HO1基因(pS/HO1)转染到人胃癌细胞系SGC-7901内,经G418筛选并建立siRNA表达载体稳定沉默胃癌SGC-7901细胞HO1基因的细胞系,分为SGC-7901-pS/HO1组,转染空质粒细胞(SGC-7901-pS)组和未处理细胞(SGC-7901)组;用实时荧光定量PCR和蛋白印迹验证HO1基因在各组细胞中的表达,并通过CCK-8和克隆形成实验分别观察HO1基因被干扰后细胞的生物学行为。结果与SGC-7901-pS组相比,SGC-7901-pS/HO1细胞中HO1基因蛋白表达明显减少,降低了5.58倍(0.321±0.051 vs 1.675±0.153,P<0.05);与对照组相比较,SGC-7901-pS/HO1实验组较SGC-7901-pS对照组细胞增殖数量明显减少(P<0.001);与转染pS空载体的SGC-7901-pS细胞对照组相比,SGC-7901-pS/HO1细胞的克隆形成明显减少,降低了3.45倍(8.32±1.142 vs 2.32±0.362,P<0.05)。结论 HO1基因真核siRNA表达载体筛选成功,为继续深入的研究HO1基因在胃癌中的功能提供了依据。 Objective To investigate the effect and function of HO1 gene in its human gastric cancer SGC-7901 cell line by constructing the eukaryotic siRNA expression vector of HO1 gene and to explore its potential role in gastric carcinogenesis.Methods A recombinant plasmid carrying siRNA specific for HO1 gene expression vector was constructed and transfected into SGC-7901 cells by lipofectin-mediated method.Cells（SGC-7901-pS/HO1） stably expressing siRNA specific for HO1 gene were selected using G418.SGC-7901 cells untransfected and those transfected with empty pS plasmid（SGC-7901-pS） were used as controls.The expression levels of HO1 gene were detected by real-time PCR and Western blot in stably transfected-SGC-7901（SGC-7901-pS/HO1） cells,biological function of HO1 gene was analyzed by cell growth activity and soft agar colony formation assay.Results Compared with SGC-7901-pS cells,the expression of HO1 protein of SGC-7901-pS/HO1 cells was an average 5.58-fold decrease（0.321±0.051 vs 1.675±0.153,P0.05）.The colony formation rate of SGC-7901-pS/HO1 cells was significantly reduced as compared to that of control group（P0.05）.The cell growth activity showed same tendency in SGC-7901-pS/HO1 group,SGC-7901-pS cells group and SGC-7901 cells group.Conclusion The eukaryotic expression vector of siRNA specific for HO1 gene was constructed successfully,which might contribute to probe the functions of HO1 in human gastric cancer in depth.
作者 程力 李乐平 张黎 苗瑞政 靖昌庆 郭晓波
机构地区 山东大学附属省立医院胃肠外科
出处 《基础医学与临床》 CSCD 北大核心 2012年第11期1259-1263,共5页 Basic and Clinical Medicine
基金 国家自然科学基金青年基金(81101858) 山东省自然科学基金(Y2007C127 ZR2011HM041)
关键词 胃癌 HO1基因 干扰载体 SGC-7901细胞 gastric carcinoma HO1 gene siRNA vector SGC-7901 cells
分类号 R735.2 [医药卫生—肿瘤]
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共引文献2

同被引文献13

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基础医学与临床
2012年 第11期

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