URI基因稳定表达对肝癌SMMC-7721细胞增殖能力的影响

中国生物工程杂志

2013, Vol. 33

Issue (6): 1-6

研究报告

URI基因稳定表达对肝癌SMMC-7721细胞增殖能力的影响

季辉, 王琦, 仲蕾, 魏文祥

苏州大学医学部基础医学与生命科学学院苏州 215123

The Effect of URI Gene Stable Expression on Cell Proliferation in SMMC-7721 Cell Line

JI Hui, WANG Qi, ZHONG Lei, WEI Wen-xiang

Dept of Cell Biology, School of Biology and Basic Medicine Science, Medical College, Soochow University, Suzhou 215123, China

全文: PDF(802 KB) HTML

摘要：

目的:瞬转以及筛选出能够稳定表达URI(RPB5-mediating protein)基因的SMMC-7721细胞株,以其为模型研究URI基因对人肝癌SMMC-7721细胞增殖的影响。方法:首先,抽提URI的重组质粒并转染到SMMC-7721细胞中,在G418药物的筛选下选出能够稳定表达URI基因的细胞株,RT-PCR法和酶切检测该稳定细胞中URI基因的表达效率以确认URI是否稳定表达,MTT法和克隆形成实验检测URI基因对SMMC-7721细胞增殖的影响。结果:成功建立URI基因过表达的稳定细胞株。与SMMC-7721细胞对照组比较,其URI mRNA表达水平显著上调,能稳定表达URI细胞株的增殖,克隆形成率明显升高。结论:pFLAG-CMV-4-URI重组质粒能使URI在肝癌SMMC-7721细胞内稳定表达,过表达URI基因有可能帮助细胞通过G2/M期检验点来提高肝癌细胞SMMC-7721的增殖能力。

关键词： 肝癌细胞; URI过表达; 细胞增殖

Abstract:

Objective: To establish the cell lines which can stably express the RPB5-mediating protein (URI) and study the effects of URI gene on the biological function of human hepatoma SMMC-7721 cells. Methods: Get PFLAG-CMV-4-URI from the E.coli which was transfected the plasmid. Then to identify the recombinant eukaryotic expression by enzyme analysis, RT-PCR Transient transfected the plasmid PFLAG-CMV-4-URI into SMMC-7721 cells by Lipofectamine reagent. G418 selected the cell clone which stably expressed URI. The URI expression were detected by RT-PCR and cell proliferation was measured by MTT assay. Plate colony forming assay was used to observe the clone formation ability of cell lines. Results: The positive clones which stably expressed URI were selected. Result of RT-PCR and enzyme analysis showed that URI gene could stably expression in SMMC-7721 cells. MTT assay showed that URI promoted the growth of SMMC-7721 cells. And the URI gene could also improve the formation rate of cell clones. Conclusion: URI gene can improve the proliferation of SMMC-7721 cells probably by helping the cell through the G2/M checkpoint.

Key words: Human hepatoma cell URI over expression Cell proliferation

收稿日期: 2012-12-10 出版日期: 2013-06-25

ZTFLH:

Q819

通讯作者: 魏文祥 E-mail: wenxiangw@suda.edu.cn

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引用本文:

季辉, 王琦, 仲蕾, 魏文祥. URI基因稳定表达对肝癌SMMC-7721细胞增殖能力的影响[J]. 中国生物工程杂志, 2013, 33(6): 1-6.

JI Hui, WANG Qi, ZHONG Lei, WEI Wen-xiang. The Effect of URI Gene Stable Expression on Cell Proliferation in SMMC-7721 Cell Line. China Biotechnology, 2013, 33(6): 1-6.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I6/1

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