论文部分内容阅读
目的 :利用外源性基因改变肿瘤原有的放射耐受性使其对放疗敏感 ,从而为提高肿瘤放射治疗疗效提供实验依据。方法 :利用本实验室构建的含腺病毒 (Ad5 )早期表达基因EIA的pCDNA3 E1A重组质粒转染肺腺癌Anip 973细胞 ,G418筛选后 ,经PCR ,RT PCR和免疫细胞化学染色鉴定 ,获得含EIA的阳性克隆。将未转染的Anip 973细胞、转染空载体质粒的Anip 973细胞和转染EIA的Anip 973细胞取 0、1、2、3、5、7、10GY剂量点 ,用6MV的X线单次照射以制作放射存活曲线 ;5GY单次照射后不同时间用MTT法测细胞存活。结果 :①转染后的阳性克隆细胞PCR ,RT PCR和免疫细胞化学染色结果说明 :EIA已整合到细胞基因组中并且稳定表达。②细胞放射存活曲线结果显示 :未转染的Anip 973细胞Do =1 14,Dq =1 5 3;空载体转染后的Anip 973细胞Do =1 13,Dq =1 44 ;EIA转染后的Anip 973细胞Do =1 0 2 ,Dq =0 8。未转染及空载体转染后的Anip 973细胞照射后细胞存活无明显差异 ,而EIA转染后的Anip 973细胞照射后细胞存活明显下降 ,存活曲线的肩区变窄 ,Dq值降低。③5GY照射后不同时间转染前后的细胞存活以 14小时最低 ,未转染的Anip 973细胞与转染EIA的Anip 973细胞 14小时存活率分别为 :6 9 5 %、41 5 %。经t检验均有显著差异。结论 :本实验的结
Objective: To use exogenous genes to alter the original radiological tolerance of tumors to make them sensitive to radiotherapy, so as to provide experimental basis for improving the efficacy of radiotherapy. METHODS: Lung adenocarcinoma cell line Anip 973 was transfected with the recombinant plasmid pCDNA3 E1A constructed with EIA in adenovirus (Ad5). The G418 was screened and identified by PCR, RT PCR and immunocytochemistry. Positive clones of EIA. The untransfected Anip 973 cells, Anip 973 cells transfected with the empty vector plasmid, and Anip 973 cells transfected with EIA were taken at doses of 0, 1, 2, 3, 5, 7, and 10 GY, using a 6MV X-ray single Irradiation was used to make a radiation survival curve; cell survival was measured by MTT assay at different times after 5GY irradiation. RESULTS : 1The results of PCR, RT PCR and immunocytochemistry staining of positive cloned cells after transfection indicated that EIA had been integrated into the cell genome and expressed stably. 2 The results of cell survival curve showed that the untransfected Anip 973 cells had Do =1 14, Dq =1 5 3; after the empty vector transfected Anip 973 cells, Do =1 13, Dq =1 44; after EIA transfection Anip 973 cells Do =1 0 2 , Dq =0 8. After irradiation with Anip 973 cells transfected with non-transfected and empty vector, there was no significant difference in cell survival. However, the cell survival was significantly decreased after irradiation with Anip 973 cells transfected with EIA. The shoulder area of the survival curve was narrowed, and the Dq value was decreased. The survival time of the cells before and after transfection with 35GY was the lowest at 14 hours. The survival rates of the untransfected Anip 973 cells and the Anip 973 cells transfected with EIA at 14 hours were 665.5% and 41.5%, respectively. The t test has significant differences. Conclusion: The knot of this experiment