| MIN Feng-ling,ZHANG Hong,LI Wen-jian,et al.Biologic effect of exogenous wild p53 combined with irradiation on human melanoma cell lines with different p53 status[J].Chinese Journal of Radiological Medicine and Protection,2007,27(2):113-116 |
| Biologic effect of exogenous wild p53 combined with irradiation on human melanoma cell lines with different p53 status |
| Received:April 17, 2006 |
| DOI： |
| KeyWords:p53 gene Adenovirus vector Irradiation Gene therapy Melanoma |
| FundProject:中国科学院2002 年百人计划基金资助项目; 科技部重大基础研究前期研究专项(2003CCB00200) |
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| Abstract:: |
| Objective To investigate the effect of low dose irradiation on gene transfer efficiency and the effect of adenoviral-mediated exogenous P53 overexpression on apoptosis and radiosensitivity of radioresistant human melanoma cell lines A375(wild type p53)and WM983a(mutant type p53). Methods Control vector, a replication deficient recombinant adenoviral vector containing a CMV promoter and green fluorescent protein (AdCMV-GFP), was used to transfect A375 cells and WM983a cells preirradiated with or without 1 Gy X-ray. The transduction efficiency of GFP gene was determined with fluorescence microscope directly. These two types of cells irradiated by 1 Gy X-ray were transfected with a replication deficient recombinant adenoviral vector carrying human wild p53 (AdCMV-p53), and mRNA level was detected by RT-PCR. The cell cycle delay and the expression of exogenous P53 were detected using flow cytometry (FCM) at different times after transfection. Tunel technique was used to detect cell apoptosis. The radiosensivity of A375 and WM983a cells after p53 transduction was analyzed by colony formation. Results It is found that 1 Gy irradiation increased the gene transfection efficiency of A375 and WM983a cells. The expression of exogenous P53 was found to range from 60% to 80% among transfected cells during the first three days after transduction and then declined continuously down to the control level on day 10. G1 cell cycle arrest was also observed after p53 gene transduction. WM983a cells transfected with p53 showed higher sensitivity to X-ray-induced cell killing than A375 cells. Conclusions It is indicated that low dose of ionizing radiation can improve gene transfection efficiency of A375 and WM983a cells mediated by adenovirus vector. Althrough the overexpresion of exogenous p53 may not inhibit cell growth and induce apoptosis of melanoma cell line A375 and WM983a in vitro, the two cell lines are much more sensitive to cell death induced by irradiation. It is suggested that p53 might be a potential gene for melanoma. |
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