中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (1): 91-94.doi: 10.3969/j.issn.1673-8225.2010.01.019

• 干细胞转基因表达 • 上一篇    下一篇

大鼠骨髓间质干细胞可以成为PEX基因稳定转染的载体细胞吗？

高 歌1，牛朝诗1，2，董永飞1，张 俊1，丁宛海1   

  1. 1安徽医科大学附属省立医院神经外科，安徽省合肥市  230001；
    2安徽省立体定向神经外科研究所，安徽省合肥市 230001
  • 出版日期:2010-01-04 发布日期:2010-01-04
  • 通讯作者: 牛朝诗，博士，主任医师，安徽医科大学附属省立医院神经外科，安徽省合肥市 230001；安徽省立体定向神经外科研究所，安徽省合肥市 230001 niuchaoshi@163.com
  • 作者简介:高 歌，男，1981年生，安徽省太和县人，汉族，2009年安徽医科大学毕业，硕士，医师，主要从事脑肿瘤方面的研究。
  • 基金资助:

    国家自然科学基金(30672166）。

Can bone marrow mesenchymal stem cells be used as PEX gene stable transfection vectors?

Gao Ge1, Niu Chao-shi1,2, Dong Yong-fei1, Zhang Jun1, Ding Wan-hai1   

  1. 1Department of Neurosurgery, Anhui Provincial Hospital, Affiliated to Anhui Medical University, Hefei   230001, Anhui Province, China;
    2Anhui Provincial Stereotactic Neurosurgical Institute, Hefei   230001, Anhui Province, China
  • Online:2010-01-04 Published:2010-01-04
  • Contact: Niu Chao-shi, Doctor, Chief physician, Department of Neurosurgery, Anhui Provincial Hospital, Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China niuchaoshi@163.com
  • About author:Gao Ge, Master, Physician, Department of Neurosurgery, Anhui Provincial Hospital, Affiliated to Anhui Medical University, Hefei 230001, Anhui Province, China
  • Supported by:

    the National Natural Science Foundation of China, No.30672166*

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摘要：

背景：PEX基因能够干预恶性胶质瘤的侵袭行为，而骨髓间质干细胞是一种靶向肿瘤治疗的新型细胞载体。

目的：建立稳定表达PEX基因的大鼠骨髓间质干细胞。

方法：采用分子克隆技术构建PEX基因真核表达载体，酶切测序鉴定重组质粒pcDNA3.1(+)-PEX，并转染大鼠骨髓间质干细胞，免疫细胞化学法验证真核表达载体在骨髓间质干细胞中的表达。通过G418筛选建立稳定表达PEX基因的骨髓间质干细胞，并用RT-PCR法检测PEX基因的表达

结果与结论：实验成功构建稳定表达PEX基因的大鼠骨髓间质干细胞，基因水平和蛋白水平检测结果表明，PEX在转染的骨髓间质干细胞中呈高表达。

关键词: PEX, 胶质瘤, 真核表达载体, 骨髓间质干细胞, 干细胞

Abstract:

BACKGROUND: PEX gene can interfere with the invasion acts of malignant glioma. Bone marrow mesenchymal stem cells (MSCs) are a new type of targeted cell vector on cancer therapy.

OBJECTIVE: To construct MSCs stably expressing PEX gene.

METHODS: PEX eukaryotic expression vector was constructed by molecular cloning, and identified the recombinant plasmid pcDNA3.1(+)-PEX by restriction endonuclease digestion and sequencing. After transfected with MSCs, the eukaryotic expression vector expression in MSCs was verified by immunocytochemical method. The MSCs stably expressing PEX was established by G418 selection, and then was detected by using reverse transcriptase-polymerase chain reaction.

RESULTS AND CONCLUSION: The MSCs stably expressing PEX gene is successfully established, in which PEX gene is highly expressed at both gene level and protein level.

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