| LIANG Nan,XIN Rui,XU Hui-ying,et al.Roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells[J].Chinese Journal of Radiological Medicine and Protection,2012,32(6):565-569 |
| Roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells |
| Received:June 20, 2012 |
| DOI：10.3760/cma.j.issn.0254-5098.2012.06.002 |
| KeyWords:dCK gene Ser-74 Irradiation Apoptosis Autophagy |
| FundProject:国家自然科学基金(30970682;30770649); 国家大学生创新项目(2010A72110) |
| Author Name | Affiliation | E-mail | | LIANG Nan | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | XIN Rui | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | XU Hui-ying | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | LIU Xiao-dong | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | GAO Bing | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | HOU Wei | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | ZHANG Xue-he | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | | | MA Shu-mei | Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China | shmm2001@yahoo.com.cn |
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| Abstract:: |
| Objective To investigate the roles of dCK Ser-74 in radiation-induced cell death in breast cancer cells. Methods Different phenotypes of dCK plasmids were transfected into MCF-7 cells by liposome transfection, including dCK-Vector, dCK-WT(wild type), dCK-S74A (non-phosphorylation) and dCK-S74E (hyper-phosphorylation). All these cells were irradiated by 0,2,4,6,8 Gy X-rays, respectively. The transcriptional and translational level of dCK were detected with real time-PCR and Western blot, respectively. Radiosensitivity was analyzed using cell counting kit (CCK-8) and colony formation assays. Monodansylcadaverine staining (MDC) and flow cytometry were used to detect autophagy and apoptosis, respectively. Results Four phenotypes of dCK cell models were established successfully. After irradiation, the cell viabilities of MCF-7 and dCK-Vector decreased significantly as compared with mock group (t=14.469 and 9.357,P＜0.05), the cell viabilities of dCK-WT, dCK-S74A and dCK-S74E showed no changes (P＞0.05). The total mortalities of dCK-WT and dCK-S74E decreased significantly as compared with dCK-Vector (χ2=3.857-3.971,P＜0.05), but no changes in dCK-S74A cells (P＞0.05). The apoptosis rates in dCK-S74A, dCK-Vector and control group were up-regulated after irradiation(t=-4.531,-3.688 and -7.076,P＜0.05), and the irradiation-induced apoptosis was reversed in dCK-WT and dCK-S74E (66% and 68% of the increase level in dCK-Vector group). The autophagy in dCK-WT and dCK-S74E increased by 22% and 26% (t=-9.051 and -8.411,P＜0.01), but no changes were observed in dCK-S74A, dCK-Vector and control groups (P＞0.05). Conclusions The dCK-WT and dCK-S74E could reverse the irradiation-induced apoptosis, increase the autophagy occurence, and decrease the total mortality, indicating that the phosphorylation of dCK at Ser-74 sites is related to the radiosensitivity of MCF-7 cells. |
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