FAN Chan,WANG Yu,LIU Xiao-dan,HUANG Bo,XU Qin-zhi,ZHOU Ping-kun.A cell co-culture model for studying bystander effect and its application on bystander DNA double-strand breaks induced by alpha-particles irradiation[J].Chinese Journal of Radiological Medicine and Protection,2013,33(3):248-251
A cell co-culture model for studying bystander effect and its application on bystander DNA double-strand breaks induced by alpha-particles irradiation
Received:January 08, 2013  
DOI:10.3760/cma.j.issn.0254-5098.2013.03.008
KeyWords:Double-strand breaks  Bystander effect  α-particle  Immunofluorescence staining  Laser confocal  γH2AX
FundProject:国家自然科学基金(81272994)
Author NameAffiliationE-mail
FAN Chan The Institute for Environmental Medicine and Radiation Hygiene, College of Public Health, The University of South China, Hengyang 421001, China  
WANG Yu 军事医学科学院放射与辐射医学研究所  
LIU Xiao-dan 军事医学科学院放射与辐射医学研究所  
HUANG Bo The Institute for Environmental Medicine and Radiation Hygiene, College of Public Health, The University of South China, Hengyang 421001, China  
XU Qin-zhi 军事医学科学院放射与辐射医学研究所  
ZHOU Ping-kun 军事医学科学院放射与辐射医学研究所 zhoupk@bmi.ac.cn 
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Abstract::
      Objective To establish an experimental model for the study of α-particle-induced bystander effect of DNA damage and investigate the characteristics of bystander DNA double-strand break (DSB).Methods The red fluorescence fusion protein of HsBrk1-RFP was used to mark the cytoplasm of one cell line to distinguish the irradiated target cells (HFS-RFP) and the non-irradiated bystander cells (HFS) in the co-culture cellular model. After α-particle irradiation, cellular DSB and its repair kinetics were analyzed by the immunofluorescence staining of γH2AX and laser confocal microscope observation. Results A bystander studying model was established by co-culturing human HFS-RFP cells with its partner HSF cells. After 0.1 Gy or 0.2 Gy α-particle irradiation, the similar kinetics of γH2AX foci production and abatement were observed in both irradiated HFS-RFP cells and non-irradiated bystander HFS cells, in which the highest level of γH2AX foci was detected at 1 h post-irradiation. The second peak of γH2AX foci formation appeared at 8 h post-irradiation, which possibly indicates the occurrence of secondary DSB. However, the production of secondary DSB in the bystander cells was weaker than that in the irradiated cells. Conclusions The cell co-culture model can be used for bystander effect investigation. Bystander DSB can be effectively induce by irradiation and the secondary breakage of DNA DSB in the bystander cells may relative to the consequential biochemical processing of clustered DNA damage.
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