Elucidation of ultraviolet radiation-induced cell responses and intracellular biomolecular dynamics in mammalian cells using surface-enhanced Raman spectroscopy

摘要

Fingerprinting biochemical changes associated with cellular responses to external stimuli can provide vital information on the dynamics of biological processes and their defense mechanisms. In this study, surface-enhanced Raman spectroscopy (SERS) has been used to elucidate biomolecular dynamics on the response of healthy and cancerous cells towards ultraviolet (UV) light irradiation at the cellular level in real-time. We have identified a number of physiochemical damages to proteins, especially to the chemical structure of the sulfur and aromatic amino acid containing moieties, as well as changes in secondary structure. Furthermore, we found that continuous exposure of short wave UV-C light (254 nm) to living cells can photolytically damage intracellular proteins and can completely arrest nanoparticle transport and trigger apoptosis. However, under similar conditions, this was not observed when the cells were exposed to long wave UV-A light (365 nm). These biomolecular events were probed in real-time using SERS and dark-field (DF) imaging. Specifically, this technique has been utilized for the real-time evaluation of a unique cellular defense mechanism in cancer cells towards UV exposure. Our technique provides a powerful approach to understand the mechanisms of UV light-triggered cell death, protein dynamics, and enhanced cell repair and defense machinery within cancer cells through actively monitoring molecular vibrations.