Imaging of Ras/Raf activity induced by low energy laser irradiation in living cell using FRET

Fang Wang; Tong-Sheng Chen; Da Xing

doi:10.1117/12.572787

18 January 2005 Imaging of Ras/Raf activity induced by low energy laser irradiation in living cell using FRET

Fang Wang, Tong-Sheng Chen, Da Xing

Proceedings Volume 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II; (2005) https://doi.org/10.1117/12.572787
Event: Photonics Asia, 2004, Beijing, China

Abstract

Ras/Raf signaling pathway is an important signaling pathway that governs cell proliferation, differential and apoptosis. Low-energy laser irradiation (LELI) was found to modulate various processes. Generally, cell proliferation is induced by low doses LELI and apoptosis is induced by high doses LELI. Mechanism of biological effect of LELI has not been clear. Recently, activation of MEK (mitogen-activated protein kinase) and ERK (extracellular-signal-regulated kinase), which are downstream protein kinases of Ras/Raf, are observed during LELI-induced cell proliferation by immunoprecipitation and western blot analysis. RaichuRas reporter consisting of fusions of H-ras, the Ras-binding domain of Raf (RafRBD), a cyan fluorescent protein (CFP) and a yellow fluorescent protein (YFP). Therefore, intramolecular binding of GTP-Ras to RafRBD brings CFP close to YFP and increases FRET between CFP and YFP. Human lung adenocarcinoma cell line (ASTC-a-1) was transfected with the plasmid (pRaichuRas) and then treated with LELI at dose of 60J/cm². Effect of LELI on Ras/Raf in physiological condition of living cells was observed by fluorescence resonance energy transfer (FRET) technique during lung adenocarcinoma cell apoptosis induced by high dose (60J/cm²) LELI. Experimental results showed that after high dose LELI treatment, the binding of Ras and Raf decreases obviously, Ras/Raf signaling pathway deregulates and cell apoptosis occurs.

Citation Download Citation

Fang Wang, Tong-Sheng Chen, and Da Xing "Imaging of Ras/Raf activity induced by low energy laser irradiation in living cell using FRET", Proc. SPIE 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II, (18 January 2005); https://doi.org/10.1117/12.572787

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