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目的探讨鼻咽癌细胞不同p53功能状态及其放射生物学特性之间的关系。方法采用脂质体介导的转染方法将携带野生型p53基因的真核表达质粒pC53SN3,空载体质粒pCMVNeoBam分别转染到鼻咽癌细胞CNE1、CNE2中。p53功能检测技术明确细胞转染p53基因前后的p53功能状态。成克隆实验测定细胞存活分数。采用单击多靶模型和线性二次函数模型拟合细胞存活曲线,求出放射生物学参数D0、Dq、N和α、β、α/β、SF2值。结果转染野生型p53基因后鼻咽癌细胞获得正常p53功能。CNE1pNeo和CNE1wtp53细胞的D0值分别为1.17、1.08Gy;Dq值分别为2.25、1.21Gy;α值分别为0.13、0.29Gy1;SF2值分别为0.765、0.326。CNE2pNeo和CNE2wtp53细胞的D0值分别为0.92、0.84Gy;Dq值分别为1.45、1.04Gy;α值分别为0.13、0.76Gy1;SF2值分别为0.675、0.156。CNE1、CNE2细胞转染野生型p53基因后,D0、Dq、SF2值均减小,α值增大。结论野生型p53基因转染可以提高p53功能缺陷的鼻咽癌细胞的放射敏感性。
Objective To investigate the relationship between the functional status of different p53 and the radiobiological characteristics of nasopharyngeal carcinoma cells. Methods The eukaryotic expression plasmid pC53SN3 carrying the wild-type p53 gene and the empty plasmid pCMVNeoBam were transfected into CNE1 and CNE2 nasopharyngeal carcinoma cells by liposome-mediated transfection. p53 function detection technology to clear p53 functional status of p53 cells before and after transfection. Into cloning experiments to determine cell survival score. The cell survival curves were fitted by single-click multi-target model and linear quadratic function model, and the radiobiological parameters D0, Dq, N and α, β, α / β and SF2 values were obtained. Results After transfection of wild type p53 gene, nasopharyngeal carcinoma cells obtained normal p53 function. The D0 values of CNE1pNeo and CNE1wtp53 cells were 1.17 and 1.08Gy respectively, the Dq values were 2.25 and 1.21Gy, the alpha values were 0.13 and 0.29Gy1, and the SF2 values were 0.765 and 0.326, respectively. The D0 values of CNE2pNeo and CNE2wtp53 cells were 0.92 and 0.84 Gy, respectively; the Dq values were 1.45 and 1.04 Gy, the alpha values were 0.13 and 0.76 Gy, respectively; the SF2 values were 0.675 and 0.166, respectively. After transfection of CNE1 and CNE2 cells with wild-type p53 gene, D0, Dq, SF2 values decreased and α value increased. Conclusion Transfection of wild-type p53 gene can enhance the radiosensitivity of NPC-deficient NPC cells.