慢病毒介导的酸性成纤维细胞生长因子基因在乳腺癌细胞MCF-7中的表达及其意义

J4 ›› 2012, Vol. 38 ›› Issue (1): 46-49.

慢病毒介导的酸性成纤维细胞生长因子基因在乳腺癌细胞MCF-7中的表达及其意义

王伟,施恒亮,付春玲,曹让娟,朱筱娟

（东北师范大学遗传与细胞研究所神经可塑性实验室,吉林长春 130024）

收稿日期:2011-06-27 出版日期:2012-01-28 发布日期:2012-01-28
通讯作者: 朱筱娟 E-mail:（Tel: 0431-85099769,E-mail: zhuxj720@nenu.edu.cn）
作者简介:王伟（1987-）|女|山东省济南市人|细胞生物学硕士|主要从事肿瘤遗传及细胞学的研究。
基金资助:
教育部科学技术研究重点项目资助课题（107036）

Expression of FGF-1 gene in breast cancer cells MCF-7 |mediated by lentiviral vector  and its significance

WANG Wei,SHI Heng-liang,FU Chun-ling,CAO Rang-juan,ZHU Xiao-juan

(Laboratory of Neural Plasticity,Institute of Genetics and Cytology,Northeast Normal University,Changchun 130024,China )

Received:2011-06-27 Online:2012-01-28 Published:2012-01-28

摘要/Abstract

摘要：

目的：探讨慢病毒系统介导的实现酸性成纤维细胞生长因子 (FGF-1) 在 MCF-7 细胞中的过表达，获得初步稳定并且大量表达 FGF-1 的细胞株，为研究 FGF-1 在肿瘤发生及发展中的作用奠定基础。方法：构建 FGF-1的慢病毒表达载体 pWPXLd-FGF-1，通过与包装质粒 psPAX2、 pMD2.G 共转染 293FT 细胞进行病毒包装，用包装成功后病毒液侵染乳腺癌 MCF-7 细胞，通过流式细胞仪检测侵染效率，通过 Western blotting 检测 GFP-FGF-1 的融合表达,通过 Wound healing 实验初步检测 FGF-1 对细胞增殖及迁移能力的影响。 结果：成功 构建了FGF-1的慢病毒表达载体，病毒滴度达到2.8×10⁶ TU•mL^-1；获得一株大量表达 FGF-1 的细胞株，FGF-1的转染效率达60%以上，FGF-1对细胞的增殖及迁移具有一定的促进作用。结论：成功构建FGF-1的慢病毒表达载体并且实现在 MCF-7 细胞中的大量表达。

关键词: 酸性成纤维细胞生长因子；慢病毒属；乳腺肿瘤；伤口愈合

Abstract:

Abstract:Objective To study the expression of acidic fibroblast growth factor(FGF-1) in MCF-7 cells mediated by lentivirus system,and obtain a primarily stable cell line highly expressing FGF-1 and to lay foundation for further exploring the role of FGF-1 in the tumorigenesis and development of breast cancer. Methods The lentivirus-expressing plasmid pWPXLd-FGF-1 was constructed and cotransfected into 293FT cells with lentiviral vector psPAX2,pMD2.G and pWPXLd-FGF-1. The lentivirus were collected and infected MCF-7 cells,the expression of FGF-1 was tested by flow cytometry and Western blotting. The capabilities of proliferation and migration of MCF-7 cells transfected with FGF-1 were also examined by wound healing test. Results The pWPXLd-FGF-1 plasmid was successfully constructed. The lentivirus was obtained and the titer of lentivirus reached to 2.8×10⁶ T U•mL^-1. In addition,a cell line highly expressing FGF-1 was obtained,its transfection efficiency was more than 60%,which promoted the proliferation and migration of MCF-7 cells. Conclusion A lentivirus vector of FGF-1 is sucessfully constructed,and it can higly express in MCF-7 cells.

Key words: acidic fibroblast growth factor;lentivirus;breast neoplasms;wound healing

中图分类号:

Q813.11

王伟,施恒亮,付春玲,曹让娟,朱筱娟. 慢病毒介导的酸性成纤维细胞生长因子基因在乳腺癌细胞MCF-7中的表达及其意义[J]. J4, 2012, 38(1): 46-49.

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