Raman-Based in Situ Monitoring of Changes in Molecular Signatures during Mitochondrially Mediated Apoptosis

摘要

Obtaining molecular information from inside cells is an important topic to understand the outcome of molecular interactions between potential drug molecules and biomolecules inside cells. To envision this goal, we investigated the surface-enhanced Raman scattering-based single-cell spectroscopic method to monitor changes in intracellular molecular signatures during mitochondrially mediated apoptosis in real time. Triphenylphosphine-modified gold nanoparticles were localized successfully to the mitochondria and greatly enhanced to obtain the intrinsic Raman scattering spectrum of mitochondria and cytochrome c in the live cell. Photothermally induced apoptosis showed a moderate decrease in the disulfide bond and a sharp increase in β-sheet structures depending on the input-laser power, along with morphological changes. However, chemical drug induced-apoptosis showed more subtle changes in the disulfide bond, as well as changes in Raman peaks corresponding to cytochrome c, and the appearance of a new peak at 1420 cm–1, which enabled us to study the molecular interactions within the mitochondria in real time from a single cell, following treatment with a novel pyruvate dehydrogenase kinase inhibitor.