Mechanism study of low-energy laser irradiation-induced lung adenocarcinoma cell proliferation by FRET in living cell

Fang Wang; Xiao-Chuan Chen; Da Xing

doi:10.1117/12.572048

29 July 2004 Mechanism study of low-energy laser irradiation-induced lung adenocarcinoma cell proliferation by FRET in living cell

Fang Wang, Xiao-Chuan Chen, Da Xing

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Proceedings Volume 5486, ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics; (2004) https://doi.org/10.1117/12.572048
Event: ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics, 2003, Silsoe, United Kingdom

Abstract

Low-energy laser irradiation (LELI) has been shown to promote cell proliferation in various cell types, yet the mechanism of which has not been fully clarified. The Ras/Raf/MEK (mitogen-activated protein kinase)ERK kinase)/ERK (extracellular-signal-regulated kinase) signaling pathway is a network that govern proliferation, differentiation and cell survival. Recent studies suggested that Ras/Raf/MEK/ERK pathway is involved in the LELI-induced cell proliferation. Here, we utilized fluorescence resonance energy transfer (FRET) technique to investigate the effect of LELI on Ras/Raf signaling pathway in living cells. Raichu-Ras reporter plasmid was utilized which consisted of fusions of H-ras, the Ras-binding domain of Raf(RafRBD), a cyan fluorescent protein (CFP) and a yellow fluorescent protein (YFP), so that 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) were transfected with the plasmid (pRaichu-Ras) and then were treated by LELI. The living cell imaging showed the increase of FRET at different time points after LELI at the dose of 1.8 J/cm², which corresponds to the Ras/Raf activation assayed by Western Blotting. Furthermore, this dose of LELI enhanced the proliferation of ASTC-a-1 cells. Taken together, these in vivo imaging data provide direct evidences with temporal or spatial resolution that Ras/Raf/MEK/ pathway plays an important role in LELI-promoted cell proliferation.

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Fang Wang, Xiao-Chuan Chen, and Da Xing "Mechanism study of low-energy laser irradiation-induced lung adenocarcinoma cell proliferation by FRET in living cell", Proc. SPIE 5486, ALT'03 International Conference on Advanced Laser Technologies: Biomedical Optics, (29 July 2004); https://doi.org/10.1117/12.572048

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KEYWORDS

Fluorescence resonance energy transfer

Lung

Radium

Fluorescent proteins

Helium neon lasers

Laser irradiation

Proteins

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