Dynamic Activation of Src Induced by Low-power Laser Irradiation in Living Cells Mediated by Reactive Oxygen Species

Src tyrosine kinases are central components of adhesive and growth factor responses and play key roles in the control of cell proliferation, survival and adhesion. In present study, to monitor Src activity in living cells in real time, we transfected Hela cells with Src reporter constructed on the basis of fluorescence resonance energy transfer (FRET) technique. We reported that 15 J/cm² low-power laser irradiation (LPLI),as well as the addition of EGF to serum-starved Hela cells, activates tyrosine kinase Src. Reactive oxygen species (ROS) as intracellular messengers , were reported to play important roles in LPLI and EGF triggered signaling pathway. Using fluorescent probes 2´,7´-dichlorodihydrofluorescein diacetate (H2DCFDA) , we found LPLI and EGF stimulation caused ROS increase to 40% and 100% above the basal level. Exogenous added H₂O₂ (200 ~ 250 muM) can also markly activate Src. We suggest that LPLI activates Src by inducing ROS production.