针对HBV X基因区三个关键位点的反义寡核苷酸体内抑瘤研究  

Study on growth inhibition of heptoma cells and anti-HBV expression of antisense oligodeoxynucleotides on HBV X gene in vitro

作　　者：曹英林[1] 刘素侠[1] 张利宁[1] 马春红[1] 丁培芳[1] 孙汶生[1] 丁红转 

机构地区：[1]山东大学医学院免疫学研究所 [2]济南市房管中心医院

出　　处：《山东大学学报（医学版）》2002年第3期193-194,198,共3页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金课题 ( 30 0 70 341)

摘　　要：目的 :研究HBVX基因区关键区段的反义寡核苷酸 (asON)对人肝癌裸小鼠模型的抑瘤生长及体内抗HBV作用。方法 :PCR法分别合成了互补于HBVX基因的翻译起始区Xp、DR2、ENⅡ区的反义寡核苷酸及无关对照序列 ,进行硫代化修饰 ,以HBVDNA转染的HepG2 .2 .15细胞接种裸小鼠 ,2 4h内同部位一次性用药 10 0 μg ,观察并比较 3种反义硫代寡核苷酸的体内抑瘤作用。ELLSA法检测反义核酸作用裸小鼠血清中HBsAg和HBeAg含量的变化。结果 :3种药物作用裸小鼠组均表现为出瘤潜伏期延长 ,出瘤率明显低于未用药瘤细胞对照组 (P <0 .0 5 ) ,且瘤体生长缓慢。互补于Xp、DR2、ENⅡ区的反义寡核苷酸一次性给药方式对荷瘤鼠HBsAg和HBeAg的表达无明显抑制作用 ;无关序列不影响荷瘤鼠瘤体生长及HBV抗原表达。结论 :针对HBVX基因区关键部位的反义寡核苷酸可抑制荷瘤裸小鼠人肝癌的生长 。Objective: To Study the growth inhibition effect of oligodeoxy- nucleotides on HBV X gene. Methods: Three antisense phosphorothioate oligodeoxy- nucleotides(asON) complementary to the initiator of Xp,DR2,ENⅡrespectively were synthesized through PCR method. Balb/c(nu/nu) mice were given asONs 100μg/mouse 24h after HepG2.2.15 cells 5 10 mouse were injected. The anti-HBV effect was assayed with ELISA. Results: In mice treated with as-Xp, as-ENII and as-DR2 treated a prolonged period of tumoregenesis as observed, but it was not found in the control group. There was no inhibition effect on the expression of HBsAg and HBeAg. Conclusion :These results demonstrated the anti-tumour activity of as-Xp, as-ENII and as-DR2 in vivo and the potential utility of short oligonucleotides as tumour cell inhibitors.

关 键 词：乙型肝炎病毒 反义寡核苷酸 小鼠 肝细胞瘤 

分 类 号：R373.2[医药卫生—病原生物学] R735.7[医药卫生—基础医学]