慢病毒介导的RNAi对人多发性骨髓瘤DKK1基因表达的抑制作用  

Inhibition Effect of Lentiviral Vector-mediated RNAi on Expression of Human DKK1 Gene in Human Multiple Myeloma

作　　者：章瑜[1] 俞康[1] 

机构地区：[1]温州医学院附属第一医院血液内科,325000

出　　处：《医学研究杂志》2010年第6期85-87,共3页Journal of Medical Research

基　　金：温州医学院5010重大项目子课题(XNK05005)

摘　　要：目的观察慢病毒表达载体介导的RNA干扰(RNAi)对人骨髓瘤细胞株U266中dickkopf1(DKK1)的抑制作用,为以DKK1基因为靶点的骨髓瘤骨病(myeloma bone disease,MBD)的基因治疗研究奠定基础。方法把构建好的DKK1shRNA慢病毒表达载体,与慢病毒包装质粒混合,以L ipofectam ine2000转染293FT细胞,48h后收取含病毒的上清液、浓缩,并检测病毒效价;RT-PCR和Western blot检测病毒感染多发性骨髓瘤细胞U266细胞后的DKK1表达。结果浓缩后的慢病毒效价为4.27×105TU/ml;DKK1shRNA慢病毒感染的U266细胞分别与未感染的U266细胞、无关序列shRNA慢病毒感染U266细胞DKK1表达量比较,统计学均有显著性差异(P<0.001)。结论慢病毒介导的干扰RNA能有效抑制骨髓瘤细胞U266中DKK1的表达。ObjectiveTo observe the inhibition effect of lentiviral vector-mediated RNA interference（RNAi）on expression of human dickkopf1（DKK1）gene in human multiple myeloma cell line U266, so as to lay the foundation for the therapeutic research of myeloma bone disease（MBD）. MethodsThe 293FT cell line was transfected by the recombined plasmid and lentivirus packing materials.The culture supernatant was harvested after 48h, and concentrated, then the virus titer was determined. RT-PCR and Western Blot were used to observe the inhibition of DKK1 expression after the lentivirus transduction in human multiple myeloma cell line U266.ResultsThe titers of concentrated virus were 4.27×105 TU/ml.DKK1 expression in U266 cells was significantly inhibited at mRNA levels as compared with the non-transfected and empty vector transfected U266 cells （P〈0.001） . ConcusionThe lentivirus-mediated shRNA can inhibit the DKK1 expression in U266 cells.

关 键 词：DKK1 RNAI 慢病毒载体 

分 类 号：R392.1[医药卫生—免疫学]