摘要
目的 研究小干扰RNA（siRNA）抑制v-ral 白血病致病因子RALA基因表达对人白血病K562细胞迁移和侵袭的影响.方法 利用LipofectamineTM 2000将化学合成的RALA siRNA转染体外培养的K562细胞,Real-time PCR检测细胞内RALA mRNA的表达水平;Western印迹检测细胞内RALA蛋白的表达水平;Boyden趋化小室实验检测细胞体外迁移和侵袭能力.结果与随机对照组相比,转染48 h后,RALA siRNA显著下调K562细胞内RALA mRNA和蛋白的表达（P＜0.05）.与随机对照组相比,转染RALA siRNA的K562细胞迁移和侵袭能力显著降低（P＜0.05）.结论 癌基因RALA在人白血病K562细胞迁移和侵袭过程中发挥重要作用,通过siRNA下调RALA的表达可抑制K562细胞迁移和侵袭能力.
Objective To investigate the effect of siRNA-induced v-ral simian leukemia viral oneogene homolog A （RALA） gene down-regulation on cell migration and invasion in human Chron- ic Myelogenous leukemia （CML） K562 cells. Methods The chemically synthesized siRNA targe- ting to RALA gene was transfected into K562 cells using LipofectamineTM 2000. The expression level of RALA mRNA and protein were determined by quantitative real-time PCR and Western blot- ting. The cell migration and invasion was examined by Boyden chamber study. Results Compared with control groups, the results of real-time PCR and Western blotting confirmed successful knock- down of RALA at mRNA and protein levels, respectively （P 〈 0.05 ） . The cell migration and invasion rates were much lower in the siRNA group than in the negative and blank control groups （P 〈 0.05 ） . Conclusion siRNA mediated knockdown of RALA results in the inhibition of migration and invasion in K562 cells. RALA might play an important role in migration of K562 cells.
出处
《医学分子生物学杂志》
CAS
CSCD
2011年第6期479-483,共5页
Journal of Medical Molecular Biology
基金
国家自然科学基金（No.81170496）
