中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (14): 2577-2582.doi: 10.3969/j.issn.1673-8225.2011.14.023

• 干细胞转基因表达 transgenic expression in stem cells • 上一篇    下一篇

构建携带胞嘧啶脱氨酶基因骨髓间充质干细胞及其对胶质瘤细胞的体外抑制作用

邢  琪1，宋  飞2，刘  健2，姬广春2，张大庆2，马郁芳3   

  1. 1大连医科大学附属第一医院麻醉科，辽宁省大连市 116011
    2大连医科大学附属第二医院神经外科，辽宁省大连市 116027
    3大连医科大学生物化学与分子生物学系，辽宁省大连市116044
  • 收稿日期:2010-09-26 修回日期:2010-12-03 出版日期:2011-04-02 发布日期:2013-11-02
  • 通讯作者: 宋飞，博士，教授，大连医科大学附属第二医院神经外科，辽宁省大连市116027 songfei168@163.com
  • 作者简介:邢琪★，女，1970年生，山东省蓬莱市人，汉族，1993年大连医科大学毕业，硕士，教授，主要从事干细胞移植的相关研究。 songfei168@163.com
  • 基金资助:

    2008年辽宁省教委科研立项(2008Z081)和大连市科学技术基金(2008E13SF203)资助，课题名称：CD/TK单、双自杀基因转染神经干细胞移植治疗脑胶质瘤的研究。2009年辽宁省自然科学基金项目(项目编号：20092165)，课题名称：胞嘧啶脱氨酶基因转染骨髓间充质干细胞移植治疗脑胶质瘤的研究。

Inhibitory effect of bone marrow mesenchymal stem cells carrying cytosine deaminase gene on glioma cells in vitro

Xing Qi1, Song Fei2, Liu Jian2, Ji Guang-chun2, Zhang Da-qing2, Ma Yu-fang3   

  1. 1Department of Anesthesia, First Hospital Affiliated to Dalian Medical University, Dalian 116011, Liaoning Province, China
    2Department of Neurosurgery, Second Hospital Affiliated to Dalian Medical University, Dalian  116027, Liaoning Province, China
    3Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • Received:2010-09-26 Revised:2010-12-03 Online:2011-04-02 Published:2013-11-02
  • Contact: Song Fei, Doctor, Professor, Department of Neurosurgery, Second Hospital Affiliated to Dalian Medical University, Dalian 116027, Liaoning Province, China songfei168@163.com
  • About author:Xing Qi★, Master, Professor, Department of Anesthesia, First Hospital Affiliated to Dalian Medical University, Dalian 116011, Liaoning Province, China
  • Supported by:

    the Scientific Research of Education Committee of Liaoning Province in 2008, No. 2008Z081*; the Scientific Technology Foundation of Dalian City, No. 2008E13SF203*; the Natural Science Foundation of Liaoning Province in 2009, No. 20092165*

PDF

569

被引次数

8

摘要：

背景：转染胞嘧啶脱氨酶基因的骨髓间充质干细胞能有效地将化疗前药5-氟尿嘧啶转化成具有细胞毒性的化疗药物5-氟尿嘧啶，并在体外对胶质瘤细胞有显著的生长抑制作用。
目的：探讨以骨髓间充质干细胞为基因治疗载体表达外源基因胞嘧啶脱氨酶基因对胶质瘤C6细胞增殖的影响。
方法：分离、培养小鼠间充质干细胞，构建胞嘧啶脱氨酶基因与GFP联合的慢病毒载体，通过慢病毒包装法将胞嘧啶脱氨酶基因及GFP转染至小鼠骨髓间充质干细胞，获得稳定表达胞嘧啶脱氨酶基因及GFP的骨髓间充质干细胞，使其与胶质瘤C6细胞共培养，在培养液中加入5-氟胞嘧啶后应用流式细胞仪检测胞嘧啶脱氨酶基因对胶质瘤细胞的增殖影响。
结果与结论：慢病毒介导的胞嘧啶脱氨酶基因及GFP基因成功转染小鼠骨髓间充质干细胞形成C57BL/6 mMSC-codA/eGFP细胞，C57BL/6 mMSC-codA/eGFP在5-氟胞嘧啶的作用下可引起胶质瘤C6细胞的明显凋亡，在5-氟胞嘧啶浓度为1×106 μg/ L条件下C6胶质瘤细胞凋亡率为60%(P < 0.05)。提示，C57BL/6 mMSC-codA/eGFP可将5-氟胞嘧啶转化成5-氟尿嘧啶并对C6胶质瘤细胞生长有显著的限制作用甚至是致死效应。

关键词: 胞嘧啶脱氨酶基因, 骨髓间充质干细胞, 胶质瘤, 慢病毒, 载体

Abstract:

BACKGROUND: The bone marrow mesenchymal stem cells (BMSCs) expressing foreign cytosine deaminase gene (gene CD) can effectively transform 5-fluorouracil (5-FC) that was used before chemotherapy into chemotherapeutics 5-FC with cytotoxicity. BMSCs expressing gene CD significantly inhibited the growth of glioma cells in vitro.
OBJECTIVE: To explore the effects of BMSCs as gene therapy vector expressing exogenous gene cytosine deaminase gene on the proliferation of glioma C6 cells.
METHODS: Mouse MSCs were isolated and cultured. Lentivirus vector combined with cytosine deaminase gene and green fluorescent protein (GFP) was constructed. Using lentivirus packaging, cytosine deaminase gene and GFP were transferred to mouse BMSCs. BMSCs expressing cytosine deaminase gene and GFP were stably obtained and cocultured with glioma C6 cells. Following 5-FC was added to the medium, effects of cytosine deaminase gene on the proliferation of glioma cells were detected using flow cytometry.
RESULTS AND CONCLUSION: Lentivirus-mediated cytosine deaminase gene and GFP gene successfully transfected mouse BMSCs and formed C57BL/6 mMSC-codA/eGFP cells. C57BL/6 mMSC-codA/eGFP could induce obvious apoptosis of glioma C6 cells following treatment with 5-FC. The apoptotic rate of C6 glioma cells was 60% (P < 0.05) under the action of 5-FC at a concentration of 1×106 μg/L. It was thus concluded that C57BL/6 mMSC-codA/eGFP could convert 5-FC to 5-FU and had significant restriction, even fatal, influence on the growth of C6 glioma cells.

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