hnRNP B1 siRNA对肺癌组织hnRNP B1表达干扰效果的体内研究

目的 在体内实验中观察所构建的核内不均一核糖核蛋白B1(hnRNP B1)特异性的siRNA真核细胞表达载体对肺癌组织hnRNP B1表达的干扰作用.方法 选取转染本课题组已构建并经体外实验证实有较好干扰效果的两对hnRNP B1特异性的siRNA真核细胞表达载体(重组质粒A和B)的人肺腺癌细胞株A549,将其接种于BALB/C nu/nu裸鼠右前腋下,构建肺癌动物模型.分别应用RT-PCR和免疫组织化学的方法检测接种40、60 d时肿瘤组织中的hnRNP B1 mRNA和蛋白质表达水平.结果 hnRNP B1特异性的siRNA真核细胞表达载体在体内表达40 d时均能较稳定而明显地抑制肿瘤组织中hnRNP B1 mRNA的表达,免疫组织化学从蛋白表达方面也予以证实,而且重组质粒A和B两组间的表达的差异无统计学意义.但是随着时间的延长(接种60 d时),siRNA的干扰效果会逐渐减弱,与未转染A549细胞比较无明显差异.结论 hnRNP B1特异性的siRNA在体内可以稳定的表达,并且能特异、高效地抑制肺腺癌细胞A549 hnRNP B1基因的表达,有望成为肺癌基因治疗的合理策略之一.

Effects of hnRNP B1-specific siRNA on the Expression of hnRNP B1 in Lung Cancer Tissues in vivo

Objective To measure the effect of the eukaryotic expression vector of siRNA specific for hnRNP B1 on the expression of hnRNP B1 in lung cancer tissues in vivo. Methods The eukaryotic expression vector of siRNA specific for hnRNP B1 was inoculated to nude mice with lung cancers. The mRNA and protein expressions of hnRNP B1 in the lung cancer tissues of the nude mice were detected by RT-PCR and Western blot. The optimal conditions for the expressions were identified. Results The eukaryotic expression vector of siRNA specific for hnRNP B1 inhibited the mRNA and protein expressions at 40 days after the inoculation. However, the inhibitive effect disappeared at 60 days after the inoculation. Conclusion The expression of hnRNP B1 gene in A549 cells can be inhibited by hnRNP B1-specific siRNA expression vectors.