| LIU Jing,WANG Ya-ting,LIN Hai,HAN Qian,ZHANG Chun-xiao,BAI Ou.The regional genomic instability induced by 60Coγ-rays in B16 cells transfected by GFP[J].Chinese Journal of Radiological Medicine and Protection,2012,32(5):465-468 |
| The regional genomic instability induced by 60Coγ-rays in B16 cells transfected by GFP |
| Received:February 07, 2012 |
| DOI：10.3760/cma.j.issn.0254-5098.2012.05.004 |
| KeyWords:Genomic instability 60Coγ-rays B16 cells |
| FundProject:国家自然科学基金(31070759);吉林省科技厅国际合作课题(20070729-6) |
| Author Name | Affiliation | E-mail | | LIU Jing | The First Hospital of Jilin University, Changchun 130021, China | | | WANG Ya-ting | The First Hospital of Jilin University, Changchun 130021, China | | | LIN Hai | The First Hospital of Jilin University, Changchun 130021, China | | | HAN Qian | The First Hospital of Jilin University, Changchun 130021, China | | | ZHANG Chun-xiao | 东北师范大学生命科学院 | | | BAI Ou | The First Hospital of Jilin University, Changchun 130021, China | oubai16@yahoo.com |
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| Abstract:: |
| Objective To detect the regional genomic instability of B16 cells treated with 60Coγ-rays by a green fluorescence protein (GFP)-based genomic instability reporting system. Methods Three groups were employed as non-transfection group, vector control group and transfection group. The GFP-marked reporter construct pCMV-EGFP2XhoI for regional genomic instability was successfully transfected into B16 cells using liposome. B16 cells were selected by screening of G418 with a series of concentrations and limiting dilution cultures to yield a single colony. B16 cells with the genomic instability report system were then irradiated by 60Coγ-rays at doses of 0, 2 and 4 Gy. The regional genomic instability of B16 cells was quantified by counting the number of cells with GFP expression. Results B-16 cell strain steadily expressing the GFP-based genomic instability reporting system was established successfully. GFP-positive B16 cells were observed at 1 d after irradiation with 60Coγ-rays at doses of 2 and 4 Gy. Positive correlations between fluorescence intensity and dose and fluorescence intensity and time were also observed. The positive expression rate of GFP followed the increased of dose (F=36.55,36.76,P<0.05) and time(t=-3.27, -3.16, -4.26, -6.11, -7.17, P<0.05), and differences between groups were significant. The positive expression rate of GFP increased significantly at 3 d, and maximum expression was observed at 5 d(2.46 ± 0.24 and 3.82 ± 0.35). The level was tending towards stability. Spontaneous GFP expression at a ratio of 1/600 000 was observed in 0 Gy group after 2 weeks of culture. Conclusions The regional genomic instability of B16 cells induced by 60Coγ-rays can be detected using a GFP-labelled genomic instability reporter system. |
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