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

Paper Details

Date Published: 18 January 2005
PDF: 7 pages
Proc. SPIE 5630, Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II, (18 January 2005); doi: 10.1117/12.572787
Show Author Affiliations
Fang Wang, South China Normal Univ. (China)
Tong-Sheng Chen, South China Normal Univ. (China)
Da Xing, South China Normal Univ. (China)

Published in SPIE Proceedings Vol. 5630:
Optics in Health Care and Biomedical Optics: Diagnostics and Treatment II
Britton Chance; Mingzhe Chen; Arthur E. T. Chiou; Qingming Luo, Editor(s)

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