摘要
目的以真核表达质粒pmU6为基础,针对bi-1基因构建在细胞内表达短发夹状RNA(shR-NA)的质粒载体,并观察它们对CNE-2Z、CNE-1、HO8910PM、HO8910细胞株生长增殖的影响。方法人工合成bi-1寡核苷酸链,退火、定向克隆入pmU6载体中产生重组质粒,将重组质粒转染到细胞中,MTT比色法观察转染试剂及质粒载体对细胞生长增殖的影响。结果与未处理组相比,bi-1shR-NA对CNE-2Z、CNE-1、HO8910PM细胞株的生长增殖有明显的抑制作用,而对HO8910细胞株的生长增殖无明显抑制作用。结论重组质粒能在细胞内表达shRNA,产生RNA干扰(RNA interference,RNAi)效应并特异性抑制靶细胞的生长增殖,为质粒介导的RNAi技术应用于鼻咽癌和卵巢癌的基因治疗提供一定的理论依据。
Objective To construct several eukaryotic plasmid expressing shRNA of gene bi-1 which de rived from the eukaryotic plasmid pmU6 and study the effect on proliferation of CNE-2Z, CNE-1, HO8910, HO8910PM. Methods Oligonucleotides of bi-1 were synthesized and inserted into the eukaryotic expression vector pmU6 in definite direction. Cells were transfected with recombinant plasmids, MTT assay was applied to evaluate proliferation of cells affected by Lipofectmine^TM 2000 and the plasmids. Results Compared with untreated group, transfected with recombinant plasmids group may inhibit proliferation of CNE-2Z, CNE-1, HO8910PM markedly, but can not inhibit proliferation of HO8910. Conclusion The shRNA expressed by the recombinant plasmid can sufficiently induced RNAi and inhibit cell proliferateion in some tumor cells, and these observations may open a path toward the use of plasmidmediated RNAi as a new tool for gene therapy research in human cancer.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2008年第5期321-324,共4页
Cancer Research on Prevention and Treatment
基金
广东省重点学科资助项目(X9307)
广东医学院青年基金资助项目(Q0523)
