FRET检测tk基因诱导ACC-M细胞凋亡  

Monitoring Cell Apoptosis of ACC-M Cells Induced by TK Gene Using FRET Technique

作　　者：熊涛[1] 陈延平[2] 

机构地区：[1]长江大学生命科学学院,湖北荆州434025 [2]华中科技大学生命科学与技术学院,湖北武汉430074

出　　处：《长江大学学报（自科版）（中旬）》2007年第4期54-57,61,共5页Journal of Yangtze University(Nature Science Edition)

基　　金：湖北省教育厅青年资助项目(Q200712005)

摘　　要：研究肿瘤细胞凋亡是进行其药物筛选的重要方法之一。利用已构建的基于Caspase酶的指示剂CD3质粒与HSV-tk/ACV系统结合,通过荧光能量共振转移(FRET)技术研究了细胞凋亡,并采用ACV药物诱导表达了TK-GFP-CD3的ACC-M肿瘤细胞和CD3的ACC-M肿瘤细胞。结果表明,FRET成像检测分析出ACV药物能够诱导转有TK基因的人涎腺癌(ACC-M)肿瘤细胞凋亡,通过对仅转有CD3质粒的ACC-M肿瘤细胞没有影响。这种技术的结合可以用于细胞的凋亡药物的筛选。Inducing apoptosis of tumor cells is one of the important strategies in tumor therapy. To study the characteristics of suicide gene system of the herpes simplex virus thymidine kinase (HSV-tk) gene in tumor cells,the dynamic characteristic of apoptosis in this system and its effect on the human adenoid cystic carcinoma line ACC-M cell,CD3 (ECFP-DVED-DsRed)-and TK-GFP co-expressing ACC-M cells was used. CD3 is a FRET-based indicator for activity of caspase3. The cell apoptosis induced by acyclovir (ACV) was monitored using fluorescence resonance energy transfer (FRET) technique. FRET from ECFP to DsRed could be detected in normal CD3 and TK-GFP co-expressing ACC-M cell,while the FRET efficiency was remarkably decreased and then disappeared during cell apoptosis.It indicated that the apoptosis of TK-GFP-expressing ACC-M cells induced by ACV was via a caspase3-dependent pathway.The apoptosis of CD3 and TK-GFP co-expressing ACC-M cells were monitored ,CD3-expressing ACC-M cells induced by ACV were initiated caspase using FRET technique. The results showed ACV could induce apoptosis of CD3 and TK-GFP co-expressing ACC-M cells,but not CD3-expressing ACC-M cells. It suggested that HSV-tk system combined with ACV was effective on inducing the apoptosis of cancer cell. The same as it was proved that FRET technique combined with CD3 probe,be promising rapid-screening potential drugs for cancer research.

关 键 词：荧光能量共振转移(FRET) HSV-tk/ACV 细胞凋亡 

分 类 号：Q819[生物学—生物工程]