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论文摘要

腺病毒介导CD/5-FC系统治疗舌癌的实验研究

A Study of Adenovirus-mediated Cytosine Deaminase/5-Fluorocytosine Suicide Gene System in the Treatment of Tongue Carcinoma Cells

作者:王安训,黄洪章

Author:

收稿日期:2002-08-25          年卷(期)页码:2002,20(04):279-

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:自杀基因,胞嘧啶脱氨酶基因,基因治疗,舌肿瘤,腺病毒,

Key words:suicidegene,cytosinedeaminase,genetherapy,tonguetumor,adenoviru,

基金项目:

本课题为中山医科大学/211工程0基金(编号99074)和中山医科大学校基金(编号072014)资助项目

中文摘要

目的:探讨胞嘧啶脱氨酶基因/5-氟胞嘧啶(CD/5-FC)自杀基因治疗系统对舌癌细胞株的治疗效果。方法:应用透射电镜、RT-PCR检测含CD基因的重组腺病毒(AdCMVCD)对舌癌细胞株Tca8113的转染及表达情况,MTT法检测AdCMVCD/5-FC系统对舌癌细胞的杀伤作用及旁观者效应,FCM检测该系统对舌癌细胞周期的影响。结果: AdCMVCD可转染入舌癌细胞的胞核并表达CD基因;AdCMVCD/5-FC系统对舌癌细胞具有强杀伤作用和旁观者效应;FCM检测结果显示舌癌细胞S期比率显著升高(P0105)。结论:AdCMVCD/5-FC自杀基因系统对舌癌细胞株具有强的杀伤作用,杀伤作用具有细胞周期 (S期)特异性。

英文摘要

Objective:The purpose of this investigation was to study the effects of replication-defective recombinant adenovirus-mediated cytosine deaminase (AdCMVCD) /5-fluorocytosine (5-FC) suicide gene system on tongue carcinoma cells.Methods:AdCMVCD was propagated in 293 tongue carcinoma cells, purified by cesium chloride gradient and titrated using tissue culture infectious dos- es-50 (TCID-50). Transmission electron microscope was used to observe the adenovirus transferred into tongue carcinoma cells (Tca8113 cell line), and reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expression of CD after infection of Tca8113 cells. The killing effects and bystander effects of AdCMVCD/5-FC system on tongue carcinoma cells were evaluated using MTTassay. The changes of cell cycles after treated with AdCMVCD/5-FC system were monitored with the flow cytometry (FCM).Results:The titration of adenovirus (AdCMVCD) was 613@109/ml plaque forming units (PFUs). Under the transmission electron microscope, the adenoviral particles were found in the nucleus of tongue carcinoma cells. The recombi- nant adenovirus expressed CD after infection of Tca8113 cells in vitro. AdCMVCD/5-FC had strong killing-effects on Tca8113 cells, and these effectswere 5-FC dose-dependent. The maximum killing efficacywas 97%. The bystander effectof this system in Tca8113 cells was good. 58% of cells were killed only when 1215% of cells were infected with AdCMVCD. Compared with the control groups,the cell treated by the AdCMVCD/5-FC accumulated in the S phase of the cell cycle (P0105).Conclusion:The killing effects and the bystander effects ofAdCMVCD/5-FC system on tongue carcinoma cells are satisfying, and the AdCMVCD/5-FC system targeted on tongue carcinoma cells in the S phase.

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