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


CONCLUSIONS
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.
Department of Urology, the First Affiliated Hospital of the Medical School of Xi-an Jiaotong University, China.
Zhi-qiang
ZQ
Wang
Jun
J
Zhao
Jin
J
Zeng
Kai-jie
KJ
Wu
Yu-le
YL
Chen
Xin-yang
XY
Wang
Luke S
LS
Chang
Da-lin
DL
He
D045744:Cell Line, Tumor; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D000077022:Survivin; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology*
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D000077022:Survivin