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Comparison of caspase-3 activation in tumor cells upon treatment of chemotherapeutic drugs using capillary electrophoresis

查看全文 作　　者：Shuang [1,2]Sha;Honglin [1,2]Jin;Xiao [1,2]Li;Jie [1,2]Yang;Ruiting [3]Ai;Jinling [1,2]Lu 高影响力作者 机构地区：[1]Britton Chance Center for Biomedical Photonics-Wuhan National Laboratory for Optoelectronics,Huazhong University of Science and Technology,Wuhan 430074,China;[2]Key Laboratory of Biomedical Photonics of Ministry of Education,Huazhong University of Science and Technology,Wuhan 430074,China;[3]China National Center for Biotechnology Development,Beijing 100036,China高影响力机构 出　　处：《Protein & Cell》索引2012年第3卷第5期,共8页高影响力期刊 基　　金：supported by the National Natural Science Foundation of China(Grant Nos.30800339 and 30800208). 摘　　要：Caspases play important roles in cell apoptosis.Meas-urement of the dynamics of caspase activation in tumor cells not only facilitates understanding of the molecular mechanisms of apoptosis but also contributes to the development,screening,and evaluation of anticancer drugs that target apoptotic pathways.The fluorescence resonance energy transfer(FRET)technique provides a valuable approach for defining the dynamics of apop-tosis with high spatio-temporal resolution.However,FRET generally functions in the single-cell level and becomes ineffective when applied in the high throughput detection of caspase activation.In the cur-rent study,a FRET sensor was combined with capillary electrophoresis(CE)to achieve a high throughput method for cellular caspase detection.The FRET-based CE system is composed of a homemade CE system and a laser source for detecting the dynamics of caspase-3 in various cells expressing sensors of caspase-3 that have been treated with anticancer drugs,such as cell cycle-independent drug cisplatin and specific cell cycle drugs camptothecin and etoposide,as well as their combination with tumor necrosis factor(TNF).A posi-tive correlation between the caspase-3 activation ve-locity and drug concentration was observed when the cells were treated with cisplatin,but cells induced by camptothecin and etoposide did not show any apparent correlation with their concentrations.Moreover,differ-ent types of cells presented distinct sensitivities under the same drug treatment,and the combination treat-ment of TNF and anticancer drugs significantly accel-erated the caspase-3 activation process.Its high throughput capability and detection sensitivity make the FRET-based CE system a useful tool for investi-gating the mechanisms of anticancer drugs and anti-cancer drug screening. 关 键 词：apoptosis CASPASE-3 fluorescence reso-nance energy transfer(FRET) capillary electrophoresis(CE)
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