硫代bFGF寡核苷酸抑制HepG2和Hep2细胞的增殖  

Phosphorothioated bFGF oligonucleotides inhibits proliferation of HepG2 and Hep2 cells in vitro

作　　者：黄红亮[1] 向军俭[1] 唐勇[1] 王宏[1] 邓宁[1] 靳英杰[1] 

机构地区：[1]暨南大学分子免疫学与抗体工程中心,广东广州510632

出　　处：《基础医学与临床》2008年第3期217-221,共5页Basic and Clinical Medicine

基　　金：中国博士后科学基金(20060390735);广州市科技计划项目(2003J1-C0171)

摘　　要：目的探讨硫代修饰对bFGF寡核苷酸抑制HepG2和Hep2细胞增殖的影响。方法设计、合成bFGF寡核苷酸,聚乙烯亚胺(jetPEI)介导其转染入HepG2和Hep2细胞,荧光显微镜观察其在细胞中的定位,流式细胞仪分析转染效率,MTT法检测细胞增殖。结果bFGF反/正义硫代寡核苷酸被jetPEI介导高效转染入细胞内,主要位于细胞核。反/正义硫代寡核苷酸均呈剂量时间依赖地抑制细胞增殖,相同剂量时正硫代寡核苷酸的抑制效率高于反义硫代寡核苷酸。相应非修饰反义寡核苷酸较其互补正义链能更有效抑制两种细胞增殖。核苷酸突变明显降低反义硫代寡核苷酸对细胞抑制,并不影响正义硫代寡核苷酸的细胞抑制率。反义硫代寡核苷酸明显降低细胞中bFGF表达。结论硫代修饰bFGF反义寡核苷酸在细胞内特异性结合核酸以反义机制抑制肿瘤细胞增殖,正义寡核苷酸则因硫代修饰于细胞内以非核酸特异性机制抑制肿瘤细胞增殖。Objective To explore effects of phosphorothioate modification on bFGF oligonucleotides inhibiting proliferation of HepG2 and Hep2 cells. Methods Oligonuleotides targeting bFGF were transfected into HepG2 and Hep2 cells with jetPEI （polyethyleneimine）. Location of FITC-labeled bFGF phosphorothioate oligonucleotides was observed through fluorescence microscopy, and the transfection efficiency was ative cell proliferation was analyzed by MTT. Results analyzed by Flow CytoMeter. The relbFGF phosphorothioate oligonuleotides were efficiently transfected into HepG2 and Hep2 cells, and mainly located in nuclei. Antisense/sense phosphorothioate oligonuleotide inhibited the growth of tumor cells in a dose and time-dependent manner, sense phosphorothioate oligonuleotide more efficiently inhibited cell proliferation than antisense phosphorothioate oligonuleotides. The inhibition efficiency of corresponding bFGF antisense phosphodiester oligonuleotide in tumor cells was higher than that of sense phos- phodiester oligonuleotides with same dose. Nucleotide mismatch in antisense duced the proliferation inhibition in tumor cells. However, inhibition of tumor phorothioate oligonuleotide was not reduced by nucleotide mismatch. Moreover, phosphorothioate oligonuleotide recell growth induced by sense phosantisense phosphorothioate oligonu-leotides was significantly reduced bFGF expression in tumor cells. Conclusion These findings suggest that phosphorothioate modification has not significant impact on bFGF antisense oligonuleotide binding to nucleic acid and inhibiting tumor cell proliferation in a antisense manner, but bFGF sense phosphorothioate oligonuleotide binding to intracellular bFGF or other proteins inhibits tumor cell proliferation.

关 键 词：BFGF 肿瘤 正/反义寡核苷酸 

分 类 号：R73-3[医药卫生—肿瘤]