| ZHOU Xin-wen,XU Ya-xiang.lyGDI expression in HeLa cells increased its sensitivity to radiation-induced apoptosis[J].Chinese Journal of Radiological Medicine and Protection,2006,26(6):554-556 |
| lyGDI expression in HeLa cells increased its sensitivity to radiation-induced apoptosis |
| Received:January 10, 2006 |
| DOI： |
| KeyWords:LyGDI HeLa cell Radiosensitization |
| FundProject:国家自然科学基金资助项目(30570548);苏州大学医学发展基金资助项目(EE126607) |
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| Abstract:: |
| Objective In order to confirm whether LyGDI has apoptotic signal transduction function and can increase the apoptotic rate of radiation-induced cell death, the lyGDI and mutant D19lyGDI gene, which constructed with the pCDNA3.1 His A, were transfected into no-endogenous lyGDI HeLa cells. Methods Transient expressions of lyGDI and D19lyGDI in HeLa cells were analyzed by Western blot using anti-mono antibody of LyGDI and Xpress tag. Cell apoptosis was assayed with Annexin V-FITC apoptosis kit. To select stable clone, the transferred HeLa cells had been maintained in G418 medium for 3 weeks, then a cell line, which stably expressed LyGDI and mutant D19lyGDI, was selected. The selected cell line was irradiated with 12 Gy 60Co γ-rays. Caspase-3 activity of the cells was determined by Western blot and cell viability by clone-forming assay after 48 hours post-irradiation culture. Results Western blot and Annexin V-FITC apoptotic analysis revealed that lyGDI and D19lyGDI transient expressions in HeLa cells induced apoptosis; Caspase-3 activity measurement and clone-forming assay showed that lyGDI increased sensitivity to radiation-induced cell apoptosis. Conclusions lyGDI performs function in apoptosis signal transduction, its expression in HeLa cells can increase the sensitivity to radiation-induced cell apoptosis. |
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