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

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

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(humanembryo 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-flurocy-tosine,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-hTER-Tp-CD对Hela细胞具有靶向杀伤作用。

Objective To discuss selective killing effects of recombinant adenovirus vectors of cytosine dearninase（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 recombi- nant 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.