大肠杆菌嘌呤核苷磷酸化酶自杀基因对人胃癌细胞的作用

doi:吉林省科技厅基金资助课题（20030424-02）

大肠杆菌嘌呤核苷磷酸化酶自杀基因对人胃癌细胞的作用

黄颖，曹岩，郑永晨

吉林大学第二医院中心实验室，吉林长春130041

收稿日期:2006-03-29 修回日期:1900-01-01 出版日期:2007-01-28 发布日期:2007-01-28
通讯作者: 郑永晨

Effects of E.coli purine nucleoside phosphorylase suicidegene therapy on gastric cancer cells

HUANG Ying, CAO Yan, ZHENG Yong-chen

Central Laboratory, Second Hospital, Jilin University, Changchun 130041, China

Received:2006-03-29 Revised:1900-01-01 Online:2007-01-28 Published:2007-01-28
Contact: ZHENG Yong-chen

摘要/Abstract

摘要： 目的：探讨大肠杆菌嘌呤核苷磷酸化酶自杀基因对胃癌细胞MKN-45的抑制及杀伤作用, 并证实旁观者效应。方法：构建真核表达载体pcDNA3.1-ePNP, 将重组载体转染MKN-45细胞获得高表达大肠杆菌嘌呤核苷磷酸化酶基因（ePNP）的MKN-45细胞克隆。在MKN-45/ePNP细胞培养物中加入前体药物MePdR, 观察MePdR对该细胞的杀伤作用及旁观者效应。结果：随MePdR浓度的增大, MKN-45/ePNP细胞的生存率逐步下降,在MePdR浓度达到1 mg•L^-1时MKN-45/ePNP细胞全部被杀灭, 而未经转染的MKN-45细胞的生存率则不受影响，MePdR对MKN-45/ePNP有显著的抑制及杀灭作用；当MKN-45/ePNP细胞占总细胞的10%时，全部细胞被杀死。结论:ePNP/MePdR 自杀基因系统对胃癌细胞有生长抑制及杀伤作用并具有旁观者效应。

关键词: 自杀基因, 旁观者效应, 大肠杆菌, 嘌呤核苷磷酸化酶

Abstract: To study the effects E.coli purine nucleoside phosphorylase suicide gene on human gastric cancer MKN-45 cells as well as the bystander effect. Methods The ePNP gene was subcloned to pcDNA3.1 to construct recombinant eukaryotic expression vector pcDNA3.1-ePNP. The recombinant plasmid was transfected into MKN-45 cells by lipofectamin method and positive cell clones were screened with G418. Then MePdR was added to these gene-modified cells and the sensitivity of the cells to MePdR was studied as well as the bystander effect. Results MePdR treatment of MKN-45/ePNP cells induced cytotoxic and antiproliferative effects in a concentration-dependent manner with 100% cell death since 1 mg•L^-1. Bystander effect was strong in vitro as 100% of tumor cells were killed by MePdR with only 10% MKN-45/ePNP cells. The gene-modified gastric cancer cells MKN-45/ePNP were sensitive to the treatment of MePdR compared with unmodified tumor cells, and also a remarkable bystander effect was observed. Conclusion The efficiency of ePNP/MePdR system and the potential feasibility of suicide gene strategy for clinical therapy of human gastric cancer are demonstrated.

Key words: suicide gene, bystander effect, Escherichia coli, purine nucleoside phosphorylase

中图分类号:

R745.2

黄颖，曹岩，郑永晨. 大肠杆菌嘌呤核苷磷酸化酶自杀基因对人胃癌细胞的作用[J]. J4, 2007, 33(1): 105-108.

HUANG Ying, CAO Yan, ZHENG Yong-chen. Effects of E.coli purine nucleoside phosphorylase suicidegene therapy on gastric cancer cells[J]. J4, 2007, 33(1): 105-108.

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