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 (P <0 0 0 1) ,G2 +M期比率下降为 0 (P <0 0 0 1) ,G1/G0 期的改变不明显 (P >0 0 5 )。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。
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 doses_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 MTT assay. 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 6 3×10\+9/ml plaque forming units (PFUs). Under the transmission electron microscope, the adenoviral particles were found in the nucleus of tongue carcinoma cells. The recombinant adenovirus expressed CD after infection of Tca8113 cells in vitro. AdCMVCD/5_FC had strong killing_effects on Tca8113 cells, and these effects were 5_FC dose_dependent. The maximum killing efficacy was 97%. The bystander effect of this system in Tca8113 cells was good. 58% of cells were killed only when 12 5% 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 (P<0 001) and dropped to zero in the G\-2+M phase (P<0 001) and didn′t change in the G\-1/G\-0 phase (P>0 05).Conclusion:The killing effects and the bystander effects of AdCMVCD/5_FC system on tongue carcinoma cells are satisfying, and the AdCMVCD/5_FC system targeted on tongue carcinoma cells in the S phase.\;
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2002年第4期279-282,共4页
West China Journal of Stomatology
基金
中山医科大学"2 11工程"基金 (编号 990 74)
中山医科大学校基金 (编号 0 72 0 14 )资助项目
