全反式维甲酸增强HSV-tk/GCV系统对晶状体上皮细胞的旁观者效应
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中国广西科学基金项目(No.桂科自0899004); 中国广西卫生厅计划基金项目(No.Z2008261)


All-trans retinoic acid enhances bystander effect of HSV-tk/GCV system in rabbit lens epithelial cells in vitro
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Science Foundation of Guangxi Zhuang Autonomous Region, China(No. 0899004); Project of Guangxi Health Department, China(No.Z2008261)

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    摘要:

    目的:体外观察兔晶状体上皮细胞N/N1003A经全反式维甲酸(all-trans retinoic acid,ATRA)诱导后,2型重组腺相关病毒(recombinant adeno-associated virus 2, rAAV2)介导的单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase, HSV-tk)/丙氧鸟苷(ganciclovir, GCV)自杀基因系统对细胞旁观者效应的影响。

    方法:应用RT-PCR法检测rAAV2介导HSV-tk基因转染N/N1003A细胞后TK基因的表达,MTT 法观察经ATRA诱导后HSV-tk/GCV系统对N/N1003A细胞旁观者效应的变化,流式细胞术检测经ATRA诱导后HSV-tk/GCV系统对N/N1003A细胞凋亡的影响。

    结果:rAAV2介导HSV-tk基因成功转染N/N1003A细胞并稳定表达。与对照组比较,ATRA诱导组可显著提高HSV-tk/GCV系统对兔晶状体上皮细胞N/N1003A的旁观者效应(P<0.01),且ATRA诱导组细胞凋亡率显著增加(P<0.01)。

    结论:ATRA可显著增强rAAV2介导的HSV-tk/GCV系统对体外培养兔晶状体上皮细胞的旁观者效应。

    Abstract:

    AIM: To investigate the effect of all-trans retinoic acid(ATRA)on the bystander effect of herpes simplex virus thymidine kinase(HSV-tk)/ganciclovir(GCV)suicide gene system mediated by recombinant adeno-associated virus 2(rAAV2)in lens epithelial cells(N/N1003A)in vitro.

    METHODS: The expression of TK gene in lens epithelial cells mediated by rAAV2/HSV-tk was detected by RT-PCR. MTT assay was used to observe the changed bystander effects of HSV-tk/GCV system in N/N1003A cells after ATRA therapy. The effects of ATRA therapy on the apoptotic rates of HSV-tk/GCV system in N/N1003A cells were investigated by flow cytometry.

    RESULTS: HSV-tk gene mediated by rAAV2 was transfected successfully into N/N1003A cells and expressed stably. The bystander effect of HSV-tk/GCV system on N/N1003A cells in ATRA treatment group was significantly improved when compared with control group(P<0.01), and the apoptotic rates of the study group were notably increased(P<0.01).

    CONCLUSION: ATRA can significantly enhance the bystander effect of HSV-tk/GCV suicide gene system mediated by rAAV2 in rabbit lens epithelial cells in vitro.

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丁芝祥,彭燕一,张文彬,等.全反式维甲酸增强HSV-tk/GCV系统对晶状体上皮细胞的旁观者效应.国际眼科杂志, 2012,12(10):1844-1846.

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  • 收稿日期:2012-07-23
  • 最后修改日期:2012-08-29
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