摘要
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
Objective:To test if EIA gene is able to increase radiosensitivity of tumor cellsMethods:pcDNA3 vector with EIA gene (pcDNA3EIA) was transfected into human lung cancer cells (Anip973),then the transfected cells were selected using G418,the positive colones were identified byPCR,RTPCR and immunocytochemical methods,Anip973 cells,and cellstransfected with pcDNA3 vector (Anip973vect cells) and transfected with pcDNA3EIA (Anip973E1Acells) were irradiated with 0GY,1GY,2GY,3GY,5GY,7GY,10GY respectively using 6MV X rayAbove three cell lines werealso given 5GY irradiation respectively,survival ratios of these cells were analyzed timely with MTT methodResults:PCR,RTPCR and immunocytochemical method confirmed that EIA gene has integrated into positive transfected cellsand stably expressedCell radiationsurvival curve showed that Do and Dq value in Anip973 cell,Anip973vect cells and Anip973E1A were 114,113,102 and 153,144,08,respectivelyThe curve of Anip973 cellsand Anip973vect cells did not show difference,but Anip973EIA cells showed narrow shoulder region,and steeply fell5GYirradiated cells showed the lowest survival at 14 hour,survival rate in Anip973 cells and Anip973EIA cells were 695%and 415%(P< 005)Conclusions:Both interphase death and proliferative death of the tumor cells were enhanced significantly by EIA geneThus,introducing EIA gene into tumor cells may be a useful approach to improve the effect of radiotherapy
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2000年第3期197-199,203,共4页
Chinese Journal of Cancer
基金
国家自然科学基金!(批准号:39770822)资助
