| 稳定表达siRNA-ARK5基因的胃癌SGC-7901细胞系的构建 |
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| 引用本文： | 李文花,王海超,雷丽恰,张月超,史立宏,张宝刚. 稳定表达siRNA-ARK5基因的胃癌SGC-7901细胞系的构建[J]. 青岛医药卫生, 2013, 45(5): 321-323,329 |
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| 作者姓名： | 李文花 王海超 雷丽恰 张月超 史立宏 张宝刚 |
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| 作者单位： | 李文花 (潍坊市第二人民医院消化内科,山东潍坊,261042)； 王海超 (潍坊医学院药理学实验室)； 雷丽恰 (潍坊医学院药理学实验室)； 张月超 (潍坊医学院药理学实验室)； 史立宏 (潍坊医学院药理学实验室)； 张宝刚 (潍坊医学院病理学教研室)； |
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| 基金项目： | 国家自然科学基金资助项目(项目编号：81072068),山东省自然科学基金(项目编号：ZR2011HL047)潍坊市科学技术发展计划项目(项目编号：201201230) |
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| 摘 要： | 目的构建稳定表达siRNA-ARK5基因的人胃癌SGC-7901细胞系,为研究ARK5在胃癌发生发展中的作用与机制奠定基础。方法把pGCsilencerU6/GFP/Neo-RNAi-ARK5重组质粒通过脂质体转染胃癌SGC-7901细胞,利用不同浓度的G418筛选获得稳定转染细胞株,挑取单克隆细胞系进行培养,应用荧光显微镜和RT-PCR技术确定是否获得了ARK5基因被稳定抑制的单克隆细胞系。结果成功挑取的4株单克隆细胞中2株细胞的ARK5基因的表达被显著抑制。结论通过siRNA干扰技术成功构建了稳定表达siRNA-ARK5基因的胃癌SGC-7901细胞系,为研究ARK5在胃癌发生发展中的作用与机制奠定了基础。
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| 关 键 词： | 胃癌 ARK5 siRNA |
Establish gastric cancer SGC-7901 cell line with stable siRNA-ARK5 gene expression |
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| LI Wen-hua,WANG Hai-chao,LEI Li-qia,ZHANG Yue-chao,SHI Li-hong,ZHANG Bao-gang. Establish gastric cancer SGC-7901 cell line with stable siRNA-ARK5 gene expression[J]. QINGDAO Medical Journal, 2013, 45(5): 321-323,329 |
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| Authors: | LI Wen-hua WANG Hai-chao LEI Li-qia ZHANG Yue-chao SHI Li-hong ZHANG Bao-gang |
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| Affiliation: | Medical University. Wei fang , Shandong 261042, China |
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| Abstract: | Objective To establish gastric cancer SGC-7901 cell lines that stably express siRNA-ARK5 for the study of the mechanism of ARK5 on the occurrence and development of gastric cancer. Methods The pGCsilencerU6/GFP/Neo-RNAi-ARK5 plasmids were transfected into gastric cancer SGC-7901 cells with lipofectamine. G418 was used to selected out stable transfection clone cells, the efficiency of transfection and ARK5 gene silencing were evaluated by RT- PCR and fluorescence microscopy. Results ARK5 gene expression was significantly inhibited in 2 of 4 selected stable clones. Conclusion We successfully established gastric cancer SGC-7901 cell line with stable siRNA-ARK5 gene expression. This cell line can provide a cell model for investigating the relationship between ARK5 and the gastric cancer malignancy. |
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| Keywords: | Lung tumor ARKS siRNA |
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