腺病毒介导的TRAIL基因对舌鳞状细胞癌细胞株的影响

目的探讨以凋亡基因TRAIL为靶基因，腺病毒（ Ad）为载体，巨细胞病毒启动子（ CMV）为启动子的重组基因治疗药物AdCMV- TRAIL诱导鳞状细胞癌细胞系TCa83凋亡的作用。方法用重组绿色荧光蛋白基因（ EGFP）的腺病毒AdCMV- EGFP为对照组，转染TCa83细胞确定重组腺病毒的转染率。在细胞转染1、3、5、7 d时，应用荧光倒置显微镜观察细胞形态，应用MTT检测细胞的存活率，流式细胞仪检测细胞凋亡率。以AdCMV- TRAIL为实验组转染TCa83细胞，采用RT- PCR方法检测细胞TRAIL的表达。结果AdCMV- EGFP在1 000 particles/cell 时转染率为100%。细胞转染AdCMV- TRAIL和AdCMV- EGFP后，随时间延长细胞存活率逐渐下降，但AdCMV- TRAIL组较AdCMV- EGFP组下降更快（ P<0.001）。AdCMV- TRAIL和AdCMV- EGFP均能诱导TCa83细胞凋亡，而AdCMV- TRAIL组引起细胞凋亡的程度明显比AdCMV- EGFP组大（ P<0.000 1）。通过RT- PCR可检测到594 bp的TRAIL基因。结论AdCMV- TRAIL在体外能抑制TCa83细胞的活性，并能促进其凋亡。

Effect of TRAIL gene in human squamous cell carcinoma cell line induced by adenovirus

Objective To study the apoptotic effect on the squamous cell carcinoma cell line TCa83 induced by recombined adenovirus vector containing TRAIL gene and CMV promoter.Methods The TCa83 cell line was firstly infected with different titre of AdCMV-EGFP containing enhanced green fluorescence protein gene(EGFP) as control,and investigated the transducing rate through fluorescence to obtain the definite titre.Then TCa83 cell line was infected with AdCMV-TRAIL in proper titre,and TRAIL gene was detected by means of RT-PCR.After TCa83 cell line was infected with AdCMV-TRAIL and AdCMV-EGFP at day 1,3,5,7,the activity of TCa83 cell line were evaluated by MTT and the apoptosis were detected by flow cytometer.Results Proper titre was of 1 000 particles/cell,and TCa83 cell line could be infected 100% in this titre.TRAIL gene was detected by RT-PCR after infected with AdCMV-TRAIL.The activity of TCa83 decreased in both groups,but the AdCMV-TRAIL group decreased more sharply than AdCMV-EGFP group(P<0.001).Both AdCMV-TRAIL and AdCMV-EGFP could lead to apoptosis of TCa83 cells,but the AdCMV-TRAIL function stronger than AdCMV-EGFP.Especially there was remarkable statistic difference between two groups(P<0.000 1).Conclusion AdCMV-TRAIL could effectively decrease the activity of TCa83 cell line and induce apoptosis.