由Egr-1调控TK基因灭活胰腺癌细胞的实验研究
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The killing effect on pancreatic cancer cell in vitro by the radio-inducible TK suicide gene that controlled by Egr-1 promoter
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    摘要:

    目的:观察对放射敏感的早期生长反应基因-1(early growth response-1,Egr-l)启动子调控单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,TK)基因是否对胰腺癌细胞的杀伤作用有增敏效应。方法:以细菌内同源重组法构建含 TK基因的腺病毒载体pAdEgr-1-TK,转染人胰腺癌细胞株PC-3,60Co-γ射线照射后,逆转录-聚合酶链反应( RT-PCR)半定量分析各不同照射剂量组TK基因的mRNA的表达。加入前药丙氧鸟苷(ganciclovir,GCV),MTT法检测其对胰腺癌细胞的杀伤作用。结果:经60Co-γ射线照射后,与对照组胰腺癌细胞(0.86 ± 0.11)相比,前药GCV可明显提高对转染pAdEgr-1-TK胰腺癌细胞的杀伤效率(0.08 ± 0.03)(P < 0.001)。结论: 由Egr-l启动子调控的TK自杀基因在γ射线作用下可以显著提高杀伤胰腺癌细胞的能力。

    Abstract:

    Objective:To observe the killing effect on pancreatic carcinoma(PC-3) cell lines by early growth response-1(Egr-1) promoter activating herpes simplex virus thymidine kinase(TK). Methods:Adenoviral vector of pAdEgr-1-TK was generated through homologous recombination in bacteria and was transfected to human PC-3.After exposured to γ-radiation by a 60Co source,hemi-quantitated by RT-PCR respectively to detect the expression of mRNA of TK in different radiation dose groups, in which 0 Gy act as control group. Then the cells were added prodrug ganciclovir(GCV),the survival rate was evaluated by MTT method. Results:After irradiation,transfected cell lines(0.08 ± 0.03) were killed by prodrug GCV at higher percentage significantly compared with control group(0.86 ± 0.11)(P < 0.001). Conclusion:It indicated that the Egr-1 promoter caused high expression of TK gene in cancer cells after exposured to 60Co-γ,which should significantly boost the killing effect on pancreatic cancer cell with the prodrug.

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刘金龙,钱 清,刘训良,郭治源,杜 青,郭仕英,李朝军.由Egr-1调控TK基因灭活胰腺癌细胞的实验研究[J].南京医科大学学报(自然科学版),2007,(11):1244-1247

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  • 收稿日期:2007-04-20
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