人端粒酶逆转录酶启动子调控的胞嘧啶脱氨酶基因对宫颈癌细胞株的选择性杀伤作用研究

doi:10.3877/cma.j.issn.1673-5250.2010.03.007

中华妇幼临床医学杂志(电子版) ›› 2010, Vol. 06 ›› Issue (03) : 172 -174. doi: 10.3877/cma.j.issn.1673-5250.2010.03.007

论著

人端粒酶逆转录酶启动子调控的胞嘧啶脱氨酶基因对宫颈癌细胞株的选择性杀伤作用研究

黄淑华, 彭芝兰, 王和, 陈晓芸, 冯炜强

511400　广州，南方医科大学附属何贤纪念医院妇科
四川大学华西第二医院妇产科

出版日期:2010-06-01

Selective Killing Effect of Cytosine Deaminase Gene Controlled by Human Telomerase Reverse Transcriptase Promoter on Cervical Cancer Cells

Shu-hua HUANG, Zhi-lan PENG, He WANG, Xiao-yun CHEN, Wei-qiang FENG

Department of Obstetrics and Gynecology, Hexian Memorial Hospital of Panyu, Southern Medical University, Guangzhou 511400, China

Published:2010-06-01

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目的

探讨由人端粒酶逆转录酶(human telomerase reverse transcriptase, hTERT)启动子驱动的含胞嘧啶脱氨酶(cytosine deaminase，CD)基因的复制缺陷型腺病毒，对宫颈癌细胞的选择性杀伤作用。

方法

将自行构建复制的缺陷型腺病毒载体Ad-hTERTp-CD，由端粒酶逆转录酶启动子调控的携带胞嘧啶脱氨酶基因的重组腺病毒，分别感染人宫颈癌细胞株Hela(研究组)及人胚肺成纤维细胞(human embryo lung fibroblast cells, hELF)(对照组)，采用不同滴度重组腺病毒及不同浓度氟胞嘧啶(5-flurocytosine，5-FC)作用于Hela细胞(研究组)后，通过MTT法，检测研究组和对照组受染细胞的存活率，观察分析腺病毒载体对两种细胞的杀伤作用。

结果

MTT法检测到研究组细胞存活率随病毒滴度和氟胞嘧啶浓度的增加，不断降低；而同样处理，对人胚肺成纤维细胞(对照组)也有部分杀伤作用，但远较Hela细胞(研究组)弱，两组比较，差异有显著意义(P<0.01)。

结论

本实验构建的复制缺陷型腺病毒Ad-hTERTp-CD对Hela细胞具有靶向杀伤作用。

关键词: 人端粒酶逆转录酶启动子, 自杀基因, 胞嘧啶脱氨酶基因, 腺病毒载体

Objective

To discuss selective killing effects of recombinant adenovirus vectors of cytosine deaminase(CD) gene using a human telomerase reverse transcriptase (hTERT) promoter-driven system on cervical cancer cells.

Methods

Hela cells(study group) and human embryo lung fibroblast (hELF) cells(control group) were separately infected with recombined adenovirus. Then different titers of recombinant adenovirus and different concentrations of 5-flurocytosine acting on Hela cells were performed to measure cell viability and killing effects of adenovirus vector on two groups.

Results

After transfection by recombinant adenovirus Ad-hTERTp-CD in vitro, and combined with 5-flurocytosine, the cell growth of Hela cancer cells(study group) were suppressed compared with human embryo lung fibroblast cells(control group) (P<0.01).

Conclusion

Recombinant adenovirus combined with 5-flurocytosine has a significant killing effect on Hela cells in vitro, but not on human embryo lung fibroblast cells.

Key words: human telomerase reverse transcriptase (hTERT), suicide gene, cytosine deaminase gene, adenovirus vector

图1　Ad-hTERTp-CD/5-FC对Hela细胞的作用

图2　Ad-hTERTp-CD/5-FC对hELF的作用

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[1]	黄淑华, 彭芝兰, 王和. 人端粒酶逆转录酶启动子调控的携带胞嘧啶脱氨酶基因腺病毒载体的构建[J/OL]. 中华妇幼临床医学杂志(电子版), 2007, 03(04): 202-202.
[2]	周华平, 彭和平, 刘启才. 腺病毒介导NK4基因和单核细胞趋化蛋白1基因共表达对胰腺癌细胞的影响[J/OL]. 中华普通外科学文献(电子版), 2013, 07(03): 165-169.
[3]	孔恒, 黄宗海. 乳腺癌的自杀基因治疗研究进展[J/OL]. 中华普通外科学文献(电子版), 2007, 01(02): 122-125.

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Selective Killing Effect of Cytosine Deaminase Gene Controlled by Human Telomerase Reverse Transcriptase Promoter on Cervical Cancer Cells