摘要
目的探讨核内不均一核糖核蛋白(hnRNP)B1调节肺腺癌细胞A549抑癌基因p53表达的作用。方法采用前期研究已构建并通过体外实验证实具有较好干扰效果的两对hnRNP B1特异性的小干扰RNA(siRNA)真核细胞表达载体(重组质粒A和D)转染的人肺腺癌细胞株A549作为实验组,对照组为转染空质粒的A549细胞和未转染任何质粒的A549细胞。各组细胞均用10μmol/L顺铂作用24h收集细胞。实时荧光定量RT-PCR检测p53 mRNA的表达,Western blot检测P53及磷酸化P53蛋白表达的变化。结果各组细胞p53 mRNA和蛋白表达量无明显差异,说明hnRNP B1特异性siRNA对A549细胞p53基因mRNA和蛋白表达无影响。转染hnRNP B1 siRNA细胞磷酸化P53蛋白表达率分别为0.87±0.06、0.75±0.06,与空质粒转染组(0.30±0.08)和未转染组(0.21±0.04)相比明显增高(P<0.01)。结论hnRNP B1 siRNA可上调肺腺癌细胞A549 P53活性,参与肺癌的发生发展。
Objective To explore the effects of hnRNP B1 on the expression of p53 in human lung adenocarcinoma cell line A549. Methods Two lines of A549 ceils transfected by hnRNP B1 specific siRNA （recombinant plasmid A and D）, which had a high transfection efficiency, were established as experimental groups. One line of A549 cells and one line of A549 ceils transfected by pure plasmid served as controls. All of the cells were collected after treated with 10 μmol/L cisplatin for 24 h. The expressions of p53 mRNA and protein were detected by real-time RT-PCR and Western blot analysis. The expression of Phospho- P53 protein was detected by western blot analysis. Results No significant differences in the expression of p53 mRNA and protein among the groups were found. The expression ratio of Phospho- P53 protein in the ceils transfected by hnRNP B1 siRNA were 0. 87±0. 06 and 0. 75±0. 06,which were significantly greater than those in the A549 cell（0. 21± 0. 04） and in the A549 cells transfected by pure plasmid （0.30±0.08） （P〈0. 01）. Conclusion hnRNP B1 specific siRNA can up regulate the activity of p53 in A549 ceils.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2008年第5期812-814,835,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(批准号30270587)
教育部博士点基金(批准号20050610056)资助
