摘要
目的:采用逆转录病毒载体介导的RNAi技术靶向抑制乳癌细胞MCF-7 E2F1 mRNA的表达.方法:构建重组的表达E2F1siRNA的逆转录病毒载体.将重组载体和空载体转染入MCF-7细胞中,共分9组:空载体组、未转染组和7个转染组.筛选出稳定转染的单克隆细胞扩大培养.应用RT-PCR方法检测转染细胞和未转染细胞E2F1 mRNA的表达情况.结果:筛选出7个转染逆转录病毒载体的单克隆,命名为SIR1～7,其E2F1 mRNA表达量分别为0.857±0.015,0.850±0.037,0.907±0.029,0.878±0.021,0.900±0.021,0.912±0.031,0.913±0.023;转染空载体和未转染的细胞E2F1 mRNA表达量分别为1.563±0.031和1.544±0.018;SIR1～7 E2F1 mR-NA表达量低于空载体及未转染组,差异有统计学意义(P＜0.05),SIR1～7 E2F1 mRNA表达均受到抑制;SIR1～7之间以及空载体转染与未转染组之间差异无统计学意义(P＞0.05).结论:逆转录病毒载体介导的RNAi技术特异、高效抑制靶基因的表达,为肿瘤基因治疗提供了新的思路和工具.
Aim： To study the effects of the retrovirus-mediated RNAi on the expression of E2F1 mRNA in breast cancer MCF-7 line cell. Methods： A recombinant retrovirus expressing short interference RNA （siRNA） was constructed and transfected into the MCF-7 cells, and then the stable transfected cells were cultured. The expression of E2F1 mRNA in ceils was assessed using RT-PCR. Results, The expression of E2F1 mRNA were 0. 857± 0. 015, 0. 850 ± 0. 037, 0. 907±0. 029, 0. 878 ± 0. 021, 0. 900 ± 0. 021, 0. 912 ± 0. 031, 0. 913 ± 0. 023 for retrovirus transfected cells in 7 groups; and 1. 563 ±0. 031 and 1. 544 ± 0. 018 for negatively transfected group and untransfected group. Compared with untransfected and negatively transfected cells, the expression of E2F1 mRNA were significantly lower （ P 〈 0.05 ） in retrovirus transfected cells,and the suppression rate of E2F1 mRNA were 100% （7/7） ; the expression of E2F1 mRNA had no statistical differences （P 〉 0.05 ） among retrovirus transfected cells and between negatively transfected and untreated cells （ P 〉 0.05）. Conclusion ： Retrovirus-mediated RNAi could suppress the expression of target gene effectively, which offers a new method for cancer therapy.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2005年第5期804-806,共3页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省科技攻关基金资助项目0324410036
