Synchronized cell attachment triggered by photo-activatable adhesive ligands allows QCM-based detection of early integrin binding

被引:27
作者
Iturri, Jagoba [1 ]
Garcia-Fernandez, Luis [1 ]
Reuning, Ute [2 ]
Garcia, Andres J. [4 ,5 ]
del Campo, Aranzazu [1 ]
Salierno, Marcelo J. [1 ,3 ]
机构
[1] Max Planck Inst Polymer Res, D-55128 Mainz, Germany
[2] Tech Univ Munich, Dept Obstet & Gynecol, Clin Res Unit, D-80290 Munich, Germany
[3] Natl Sci & Tech Res Council, Buenos Aires, DF, Argentina
[4] Georgia Inst Technol, Woodruff Sch Mech Engn, Atlanta, GA 30332 USA
[5] Georgia Inst Technol, Petit Inst Bioengn & Biosci, Atlanta, GA 30332 USA
来源
SCIENTIFIC REPORTS | 2015年 / 5卷
关键词
QUARTZ-CRYSTAL MICROBALANCE; RGD-BASED STRATEGIES; REAL-TIME; CANCER; RECOGNITION; THERAPY; PEPTIDE; AVIDITY;
D O I
10.1038/srep09533
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The Quartz Crystal Microbalance with dissipation (QCM-D) technique was applied to monitor and quantify integrin-RGD recognition during the early stages of cell adhesion. Using QCM-D crystals modified with a photo-activatable RGD peptide, the time point of presentation of adhesive ligand at the surface of the QCM-D crystal could be accurately controlled. This allowed temporal resolution of early integrin-RGD binding and the subsequent cell spreading process, and their separate detection by QCM-D. The specificity of the integrin-RGD binding event was corroborated by performing the experiments in the presence of soluble cyclicRGD as a competitor, and cytochalasin D as inhibitor of cell spreading. Larger frequency change in the QCM-D signal was observed for cells with larger spread area, and for cells overexpressing integrin alpha(v)beta(3) upon stable transfection. This strategy enables quantification of integrin activity which, in turn, may allow discrimination among different cell types displaying distinct integrin subtypes and expression levels thereof. On the basis of these findings, we believe the strategy can be extended to other photoactivatable ligands to characterize cell membrane receptors activity, a relevant issue for cancer diagnosis (and prognosis) as other several pathologies.
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页数:8
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