RNA干扰抑制nm23-M1基因表达对骨髓瘤SP2/0细胞增殖的影响  

The Effects of RNA Interference to nm23-M1 Expression on Proliferation of Myeloma Cell Line SP2/0

作　　者：叶倩[1] 陈科[1] 骆明勇[1] 董艳玲[1] 王应雄[1] 何俊琳[1] 

机构地区：[1]重庆医科大学公共卫生学院生殖生物学研究室,遗传优生教研室,重庆400016

出　　处：《细胞生物学杂志》2008年第3期367-371,共5页Chinese Journal of Cell Biology

基　　金：国家自然科学基金资助项目(No.30500054)~~

摘　　要：建立RNA干扰(RNA interference,RNAi)抑制nm23-M1基因表达的骨髓瘤SP2/0细胞株,初步探讨nm23-M1基因对小鼠骨髓瘤细胞增殖的影响。针对nm23-M1 mRNA序列设计3个小干扰RNA(small interfering RNA,siRNA)序列,分别构建表达这3个序列及阴性对照序列的pGenesil-1重组质粒,再转染小鼠骨髓瘤SP2/0细胞并经G418抗性筛选稳定表达细胞株。采用半定量RT-PCR及Western印迹检测3个重组质粒对nm23-M1 mRNA及蛋白质表达的抑制效果,然后用MTT法观察抑制nm23-M1表达对SP2/0细胞增殖的影响。结果显示,设计的3条siRNA不同程度地特异抑制骨髓瘤SP2/0细胞nm23-M1 mRNA及蛋白质的表达,其中siRNA-2抑制作用最强,其对nm23-M1 mRNA和蛋白质的抑制率分别为74.4%和62.1%;且siRNA-2抑制nm23-M1基因表达后SP2/0细胞增殖受到明显抑制(P<0.05)。本研究成功构建了pGenesil-1-nm23-M1 siRNA重组质粒,筛选出稳定抑制nm23-M1基因表达的SP2/0细胞株,提示抑制nm23-M1基因表达有抑制骨髓瘤细胞增殖的作用;为进一步研究nm23基因的生物学功能及临床应用打下了基础。To investigate the effects of non-metastasis 23-M1 （nm23-M1） gene on proliferation of myeloma cell, we established a myeloma cell line SP2/0 in which nm23-M1 expression was suppressed by RNA interference （RNAi）. Three small interfering RNAs （siRNA） targeting nm23-M1 were designed, and three pGenesil-1 plasmids which could express these siRNAs and a negative control plasmid were constructed. Then the recombinant plasmids were transfected into mouse myeloma cell line SP2/0 that clones were selected by G418 in order to establish ceil lines SP2/0 with stable expression of pGenesil-1-nm23-M1 siRNA. The expression levels of nm23-M1 mRNA and protein were detected by RT-PCR and Western blot. MTT was used to observe the effects of nm23-M1 gene on proliferation of SP2/0 cells. The results showed three siRNAs could inhibit the expression of nm23-M1 on mRNA and protein levels. Among them, siRNA-2 showed the strongest inhibition effect, with an inhibition rates of 74.4% and 62.1% for the expression of nm23-M1 mRNA and protein respectively. And the proliferation of SP2/0 cells was obviously suppressed after transfection of siRNA-2 （P〈0.05）. The study constructed pGenesil-1-nm23-M1 siRNA recombination plasmids successfully and screened cell lines SP2/0 in which nm23-M1 expression was inhibited stably. The findings suggested low expression level of nm23-M1 in SP2/0 cells inhibited cell proliferation. It provides a basis for further researching the biological functions of nm23-M1 and related clinical applications.

关 键 词：nm23-M1基因 小干扰RNA RNA干扰 骨髓瘤 

分 类 号：R733.3[医药卫生—肿瘤]