活体荧光成像监测caspase-3蛋白酶的活性

Evaluation of caspase-3 activity by live cell imaging

目的利用生物发光技术,在细胞体系中实时监测caspase-3蛋白酶的活性,为细胞凋亡研究以及进行抗凋亡药物的筛查、评价提供技术支持。方法利用萤火虫荧光素酶作为细胞内凋亡发生的报告基因,构建载体,转染细胞,药物诱导细胞凋亡,利用活体荧光成像和Western印迹检测caspase-3蛋白酶的活性。结果成功建立了一种实时、动态、方便简单的检测caspase-3蛋白酶活性的细胞模型。结论该细胞模型为细胞水平上进行细胞凋亡的研究以及抗凋亡药物的筛查提供了技术平台。

Objective To establish a cellular model using bioluminescence technology for monitoring caspase-3 activity and providing a supportive technique for research into the mechanism of cell apoptosis and evaluation of anti-apoptosis drugs. Methods Using the firefly luciferase （Fluc） as a reporter to indicate cell apoptosis, a hybrid polypeptide was con- structed, transfected into a hepal-6 cell line, followed by drug-induced apoptosis. Bioluminescence technology and West- ern-blot were then used to evaluate the activity of caspase-3. Results A timely, dynamic, simple cellular model was suc- cessfully established for evaluating the activity of caspase-3. Conclusion The living cell imaging model established by us provides a technical platform for research into the mechanism of cell apoptosis and novel anti-apoptosis drugs.