中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (10): 1827-1832.doi: 10.3969/j.issn.1673-8225.2012.10.026

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

间充质干细胞靶向转运胸苷激酶基因联合更昔洛韦杀伤鼻咽癌细胞*★

钟品能，文  忠，申聪香，王海丽，王军旗   

  1. 南方医科大学珠江医院耳鼻咽喉-头颈外科，广东省广州市  510282
  • 收稿日期:2011-11-30 修回日期:2012-02-12 出版日期:2012-03-04 发布日期:2012-03-04
  • 通讯作者: 文忠，博士，教授，南方医科大学珠江医院耳鼻咽喉-头颈外科，广东省广州市 510282 wenzhong60@163.com
  • 作者简介:钟品能★，女，1985年生，广东省惠州市人，汉族，南方医科大学在读硕士，主要从事鼻咽癌基础与临床研究工作。 zhongpinneng@163.com

Herpes simplex virus thymidine kinase expressing mesenchymal stem cells in combination with Ganciclovir in targeted-gene therapy for nasopharyngeal carcinoma  

Zhong Pin-neng, Wen Zhong, Shen Cong-xiang, Wang Hai-li, Wang Jun-qi   

  1. Department of Otolaryngology-Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China
  • Received:2011-11-30 Revised:2012-02-12 Online:2012-03-04 Published:2012-03-04
  • Contact: author: Wen Zhong, M.D., Professor, Department of Otolaryngology-Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China wenzhong60@ 163.com
  • About author:Zhong Pin-neng★, Studying for master’s degree, Department of Otolaryngology- Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China zhongpinneng@163.com
  • Supported by:

    the Science and Technology Development Program of Guangdong Province, No. 2010B031200009*

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摘要：

背景：研究证实，间充质干细胞能通过基因修饰成为肿瘤治疗的靶向载体。
目的：观察转染胸苷激酶基因的胸苷激酶-间充质干细胞联合更昔洛韦对鼻咽癌细胞的靶向杀伤作用。
方法：应用LipofectamineTM 2000将表达胸苷激酶基因的重组质粒pGL3-EGFP-胸苷激酶转染至SD大鼠间充质干细胞，观察其增殖能力。应用Transwell小室观察胸苷激酶-间充质干细胞的归巢特性；将胸苷激酶-间充质干细胞植入裸鼠，观察其致瘤性；用胸苷激酶-间充质干细胞/更昔洛韦干预人鼻咽癌细胞5-8F，观察其对细胞的杀伤作用及旁观者效应。
结果与结论：荧光显微镜观察及RT-PCR检测结果提示实验成功将胸苷激酶基因转染至间充质干细胞，CCK-8检测结果显示胸苷激酶-间充质干细胞与间充质干细胞增殖能力差异无显著性意义(P > 0.05)。Transwell小室迁移实验显示胸苷激酶-间充质干细胞具有归巢特性，裸鼠移植瘤实验显示胸苷激酶-间充质干细胞无致瘤性。CCK-8检测检测发现胸苷激酶-间充质干细胞/更昔洛韦具有旁观者效应，可明显降低5-8F的生存率(P < 0.01)。提示胸苷激酶-间充质干细胞/更昔洛韦对鼻咽癌细胞具有靶向迁移及杀伤作用，间充质干细胞可作为治疗鼻咽癌的理想靶向转运载体。
关键词：鼻咽肿瘤；间充质干细胞；基因治疗；归巢；旁观者效应
doi:10.3969/j.issn.1673-8225.2012.10.026

关键词: 鼻咽肿瘤, 间充质干细胞, 基因治疗, 归巢, 旁观者效应

Abstract:

BACKGROUND: Recent studies have confirmed that mesenchymal stem cells (MSCs) can be used as targeted-gene delivery vehicles for cancer gene therapy.
OBJECTIVE: To investigate the therapeutic effects of herpes simplex virus thymidine kinase expressing mesenchymal stem cells (TK- MSCs) in combination with Ganciclovir (GCV) on nasopharyngeal carcinoma.
METHODS: The pGL3 -EGFP-TK plasmid was constructed and transfected into Sprague-Dawley rat MSCs by Lipofectamine™ 2000. Cell proliferation was determined by CCK-8 method. Tumor-homing of TK-MSCs was analyzed by Transwell inserts. BALB/C nude mice were inoculated with TK-MSCs to observe the tumorigenicity. Human nasopharyngeal carcinoma cells 5-8F were interfered with TK-MSCs/GCV to investigate the cell viability and the bystander effect.
RESULTS AND CONCLUSION: TK gene was transfected into MSCs successfully. There was no significant difference in cell proliferation between TK-MSCs and MSCs (P > 0.05). TK-MSCs maintained their tumor-homing and had no tumorigenicity. TK-MSCs/GCV significantly inhibited the growth of 5-8F cells (P < 0.01). TK-MSCs/GCV suicide gene therapy system exhibits significant inhibitory effects on growth of 5-8F cells, suggesting that MSCs can be used as delivery vehicles for targete-gene  therapy of nasopharyngeal carcinoma.

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