AKR1B10基因过表达载体的构建及其在乳腺癌MCF-7细胞中的表达

J4 ›› 2012, Vol. 38 ›› Issue (3): 465-470.

AKR1B10基因过表达载体的构建及其在乳腺癌MCF-7细胞中的表达

何鑫^1|2|李泽中²|张西臣²|刘素菊¹|邢沈阳²|宫鹏涛²|李建华^2,张国才²|陈玉江²|倪劲松¹

1.吉林大学白求恩医学院病理生物学教育部重点实验室|吉林长春 130021；2.吉林大学畜牧兽医学院|吉林长春 130062

收稿日期:2011-12-14 出版日期:2012-05-28 发布日期:2012-05-28
通讯作者: 倪劲松（Tel：0431-84886631，E-mail：nijs@jlu.edu.cn） E-mail:nijs@jlu.edu.cn
作者简介:何鑫（1984-）|女|黑龙江省齐齐哈尔市人|在读医学硕士|主要从事肿瘤病理生物学研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题(2003055025，20106044)；国家973项目资助课题(2006CB910505，2010CB530004)

Construction of overexpression vector of AKR1B10 gene and expression in breast cancer MCF-7 cells

1.Key Laboratory of Pathobiology,Ministry of Education,Norman Bethune College of Medicine,Jilin University,Changchun 130021,China|2.School of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China

Received:2011-12-14 Online:2012-05-28 Published:2012-05-28

摘要/Abstract

摘要：

目的：构建AKR1B10基因真核表达载体pcDNA3.1(+)-AKR1B10，观察其在乳腺癌MCF-7细胞中的表达，为研究AKR1B10基因与乳腺癌的关系提供实验依据。方法：构建真核表达载体pcDNA3.1(+)-AKR1B10，将载体瞬时转染乳腺癌MCF-7细胞，利用RT-PCR与Western blotting方法检测未转染的MCF-7细胞、转染pcDNA3.1(+)-AKR1B10的MCF-7细胞和转染空质粒pcDNA3.1(+)的MCF-7细胞中AKR1B10基因的表达。结果：成功构建真核表达载体pcDNA3.1(+)-AKR1B10。RT-PCR结果显示，将未转染的MCF-7细胞中AKR1B10基因表达量设定为1，转染质粒的MCF-7细胞中AKR1B10基因表达量为1.79，转染空质粒的MCF-7细胞表达量为0.98，转染表达载体细胞AKR1B10基因表达量与转染空质粒的细胞和未转染的细胞比较，差异有统计学意义（P＜0.05）；Western blotting结果显示,与未转染的MCF-7细胞和转染空质粒的细胞比较，转染表达载体pcDNA3.1(+)-AKR1B10的MCF-7细胞中AKR1B10基因表达量增多。结论：成功构建真核表达载体pcDNA3.1(+)-AKR1B10，AKR1B10基因在转染表达载体的MCF-7细胞中高表达。

关键词: AKR1B10；真核表达载体；转染；乳腺肿瘤

Abstract:

Objective
To constructed an eukaryotic expression vector pcDNA3.1(+)-AKR1B10 and observe its expression in breast cancer MCF-7 cells,and to provide experimental basis for study on the relationship between AKR1B10 gene and breast cancer.Methods The breast cancer MCF-7 cells were transfected transiently with the constructed eukaryotic expression vector pcDNA3.1(+)-AKR1B10. The expressions of AKR1B10 gene in the untransfected MCF-7 cells,the MCF-7 cells transfected with pcDNA3.1(+)-AKR1B10 and the MCF-7 cells transfected with pcDNA3.1(+) (control vector) were detected by RT-PCR and Western blotting methods.Results The eukaryotic expression vector pcDNA3.1(+)-AKR1B10 was successfully constructed.The RT-PCR results showed that if the AKR1B10 gene expression in the untransfected MCF-7 cells was regarded as 1, the expression values were 1.79 in the cells transfected with pcDNA3.1(+)-AKR1B10 and 0.98 in the cells tranfected with pcDNA3.1(+). The expression of AKR1B10 gene in the cells transfected with pcDNA3.1(+)-AKR1B10 was significantly increased compared with the cells transfected with pcDNA3.1(+) and the untransfected cells (P<0.05).The Western blotting results showed that the expression of AKR1B10 gene in the cells transfected with pcDNA3.1(+)-AKR1B10 was significantly increased compared with the cells transfected with pcDNA3.1(+) and the untransfected cells.Conclusion The eukaryotic expression vector pcDNA3.1-AKR1B10 is successfully constructed.The AKR1B10 gene highly expresses in the MCF-7 cells transfected with the expression vectror.

Key words: AKR1B10, overexpression vector, transfection

中图分类号:

R394.2

何鑫|李泽中|张西臣|刘素菊|邢沈阳|宫鹏涛|李建华,张国才|陈玉江|倪劲松. AKR1B10基因过表达载体的构建及其在乳腺癌MCF-7细胞中的表达[J]. J4, 2012, 38(3): 465-470.

HE Xin|LI Ze-zhong|ZHANG Xi-chen,LIU Su-ju|XING Shen-yang|GONG Peng-tao. Construction of overexpression vector of AKR1B10 gene and expression in breast cancer MCF-7 cells[J]. J4, 2012, 38(3): 465-470.