Specific survivin dual fluorescence resonance energy transfer molecular beacons for detection of human bladder cancer cells  

作　　者：Zhi-qiang WANG Jun ZHAO Jin ZENG Kai-jie WU Yu-le CHEN Xin-yang WANG Luke S CHANG Da-lin HE 

机构地区：[1]Department of Urology, the First Affiliated Hospital of the Medical School of Xi-an Jiaotong University, Xi-an 710061, China [2]Department of Urology, the Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, China [3]Oncological Research Lab, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi-an Jiaotong University School of Medicine, Xi-an 710061, China

出　　处：《Acta Pharmacologica Sinica》2011年第12期1522-1528,共7页中国药理学报（英文版）

基　　金：We thank Prof Xiao-hong FANG from the Institute of Chemistry, Chinese Academy of Sciences, for her helpful support. This work was supported by the National Natural Science Foundation of China （No 30672102）.

摘　　要：Aim： Survivin molecular beacons can be used to detect bladder cancer cells in urine samples non-invasively. The aim of this study is to improve the specificity of detection of bladder cancer cells using survivin dual fluorescence resonance energy transfer molecular beacons （FRET MBs） that have fluorophores forming one donor-acceptor pair. Methods： Survivin-targeting dual fluorescence resonance energy transfer molecular beacons with unique target sequences were designed, which had no overlap with the other genes in the apoptosis inhibitor protein family. Human bladder cancer cell lines 5637, 253J and T24, as well as the exfoliated cells in the urine of healthy adults and patients with bladder cancer were examined. Images of cells were taken using a laser scanning confocal fluorescence microscope. For assays using dual FRET MBs, the excitation wavelength was 488 nm, and the emission detection wavelengths were 520±20 nm and 560±20 nm, respectively. Results： The human bladder cancer cell lines and exfoliated cells in the urine of patients with bladder cancer incubated with the survivin dual FRET MBs exhibited strong fluorescence signals. In contrast, no fluorescence was detected in the survivin-negative human dermal fibroblasts-adult （HDF-a） cells or exfoliated cells in the urine of healthy adults incubated with the survivin dual FRET MBs. Conclusion： The results suggest that the survivin dual FRET MBs may be used as a specific and non-invasive method for early detection and follow-up of patients with bladder cancer.Aim： Survivin molecular beacons can be used to detect bladder cancer cells in urine samples non-invasively. The aim of this study is to improve the specificity of detection of bladder cancer cells using survivin dual fluorescence resonance energy transfer molecular beacons （FRET MBs） that have fluorophores forming one donor-acceptor pair. Methods： Survivin-targeting dual fluorescence resonance energy transfer molecular beacons with unique target sequences were designed, which had no overlap with the other genes in the apoptosis inhibitor protein family. Human bladder cancer cell lines 5637, 253J and T24, as well as the exfoliated cells in the urine of healthy adults and patients with bladder cancer were examined. Images of cells were taken using a laser scanning confocal fluorescence microscope. For assays using dual FRET MBs, the excitation wavelength was 488 nm, and the emission detection wavelengths were 520±20 nm and 560±20 nm, respectively. Results： The human bladder cancer cell lines and exfoliated cells in the urine of patients with bladder cancer incubated with the survivin dual FRET MBs exhibited strong fluorescence signals. In contrast, no fluorescence was detected in the survivin-negative human dermal fibroblasts-adult （HDF-a） cells or exfoliated cells in the urine of healthy adults incubated with the survivin dual FRET MBs. Conclusion： The results suggest that the survivin dual FRET MBs may be used as a specific and non-invasive method for early detection and follow-up of patients with bladder cancer.

关 键 词：molecular beacons fluorescence resonance energy transfer SURVIVIN human bladder cancer 

分 类 号：Q51[生物学—生物化学] Q255