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目的 观察经放射前后的转基因细胞Tca8113/TNF α的TNF α的分泌情况 ,为转基因细胞经致死剂量的放射线灭活作为“瘤苗”应用于临床前TNF α基因的有效表达提供依据。方法 利用基因转染技术 ,用逆转录病毒载体将肿瘤坏死因子基因 (TNF α基因 )转导入人舌癌Tca8113细胞 ,并获得表达 ;在此基础上 ,通过ELISA法分析转基因细胞Tca8113/TNF α在60 Co放射组及未放射组的TNF α分泌量的变化以及经液氮储存 4 8h、一周复苏后的TNF α分泌量的变化。结果 (1) 60 Co放射组转基因细胞Tca8113/TNF α的TNF α的分泌量 (x±s) (pg/ml/ 10 6细胞 /2 4h)平均高达 (110± 5 6 2 ) ,而未转基因的Tca8113细胞的上清液中未检测到TNF α ,统计学上有明显的差异性 (P <0 0 1) ;(2 )经频率为 4 39 5cGy/min ,总剂量为 10 0 0 0cGy的60 Co放射后TNF α分泌量降低 ,但在 4～ 6d时有一分泌高峰 ;(3)转基因细胞Tca8113/TNF α经60 Co放射后在经 4 8h及一周的液氮内储存后TNF α能够分泌 ,其分泌量的变化趋势同储存前。 4 8h组和一周组之间无明显的统计学差异性。结论 转基因细胞Tca8113/TNF α作为“瘤苗”应用时 ,目的基因的表达可初步评价为有效
Objective To observe the secretion of TNFα by Tca8113 / TNFα transgenic mice before and after radiotherapy, and to provide a basis for the effective expression of tumor-suppressor TNFα gene by “radiation-killed” transgenic cells. Methods Tumor necrosis factor gene (TNFα) gene was transduced into Tca8113 human tongue cancer cell by retroviral vector by gene transfection technique. The expression of Tca8113 / Tα was detected by ELISA The change of TNFα secretion in 60Co radiotherapy group and non-radiotherapy group, and the change of TNFα secretion after one week recovery after liquid nitrogen storage for 48h. Results (1) The secretion of TNFα in Tca8113 / TNFα transgenic mice was up to (110 ± 5 6 2) (pg / ml / 106 cells / 24 h) TNFα was not detected in the supernatant of Tca8113 cells (P <0.01). (2) After a total dose of 4 39 5 cGy / min and a total dose of 10 0 0 cGy TNFα secreted by 60Co after radiotherapy decreased but secreted at 4 ~ 6d; (3) TNFα secreted by Tca8113 / TNFα after 60Co radiation in 4h and 1 week liquid nitrogen , The trend of its secretion changes with the storage before. 4 8h group and no significant difference between the one-week group. Conclusion The expression of the target gene can be initially evaluated as effective when Tca8113 / TNFα is used as a “tumor vaccine”