Application of a fluorescence resonance energy transfer (FRET)-based biosensor for detection of drug-induced apoptosis in a 3D breast tumor model

Biotechnology and Bioengineering
Padmaja AnandKathy Qian Luo

Abstract

Two-dimensional (2D) cultures are commonly used for testing drug effects largely because of their easy maintenance. But they do not represent the spatial interactions of the cells within a tumor. Three-dimensional (3D) cultures can overcome those limitations thus mimicking the architecture of solid tumor. However, it is not easy to evaluate drug effects in 3D culture for a long time. This necessitates the development of a real-time and longitudinal analysis of 3D platforms. In this study, we transfected the plasmid DNA encoding the fluorescence resonance energy transfer (FRET)-based biosensor into human breast cancer cells and generated two cell lines of MCF7-C3 and MDA-MB-231-C3 (231-C3) cells. We used them to determine the activation of caspase-3, whereby healthy cells appear green and apoptotic cells appear blue by FRET imaging. As the caspase sensors can be constantly produced within the cells and quickly respond to caspase activation, we hypothesized that these sensor cells will allow longitudinal detection of apoptosis. MCF7-C3 and 231-C3 spheroids were generated and subjected to histological examination, gene expression studies, drug treatment, and FRET analyses. Our results demonstrated that MCF7-C3 cells formed tight 3...Continue Reading

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Citations

Oct 27, 2015·Sensors·Bernhard HochreiterJohannes A Schmid
Apr 9, 2019·Assay and Drug Development Technologies·Abdulaziz S FakhouriJennifer L Leight
Aug 18, 2018·Breast Cancer Research and Treatment·Sagar RegmiKathy Qian Luo
Mar 23, 2021·Journal of the American Chemical Society·Ruo-Can QianYi Lu

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