摘要
目的:探讨互补于HBVX基因的翻译起始区(as-Xp)、ENⅡ(as-ENII)区的反义寡核苷酸抑制HepG2.2.15细胞生长及抗病毒作用。方法:合成as-Xp、as-ENⅡ,体外作用于HBVDNA转染的HepG2.2.15细胞,利用ELLSA法检测反义核酸作用前后细胞上清中HBsAg和HBeAg的含量。以HepG2.2.15细胞接种裸鼠制备人肝癌模型,瘤内给予反义寡核苷酸100μg,连续给药5d,取瘤组织采用流式细胞仪(FCM)分析反义核酸诱导瘤细胞凋亡作用,并检测反义核酸作用裸鼠血清中病毒抗原表达情况。结果:as-Xp、as-ENII作用72h的细胞凋亡峰值分别为24%和27.7%;对照为10.89%、7.92%;对荷瘤鼠体内HBsAg和HBeAg的抑制率分别为35%、34%和43%、49%,在体外分别为57%、52%和56%、56%。无关对照序列于体内、外无明显的抑制瘤细胞生长和抗病毒作用。结论:as-Xp、as-ENⅡ体内应用可诱导瘤细胞凋亡,并可有效抑制体内、外HBV抗原表达。
Objective:To study the anti-HBV effect and growth inhibition of oligodeoxy-nucleotides com ple mentary to the initiator of X,ENⅡgene.Methods:as-Xp and as-ENⅡwere syn thesized through PCR method and the content of HBsAg and HBeAg was detected with ELISA after HepG2.2.15cells were giv en asON.BALB/c(nu/nu)mice were injected with HepG2.2.15cells.Once tumors were induced,the animals were administered as-Xp?as-ENⅡand saline,respectively,each mouse being given100μg /day.After5days,the tumors were examined for apoptosis through FCM.Results:In vitro,the inhibition rates of as-Xp and as-ENⅡon HBsAg were57%and52%,on HBeAg were56%and56%respectively.There were two in hibition peaks at different times.There was no inhibition effect in the control group.In vivo,apoptosis was observed in animals given asON,but not found in saline or random con trol animals.The apoptosis peaks were24%and27.7%72h after treatment.Conclusion:as-Xp and as-ENⅡcould inhibit expression of HBV,induce apoptosis,and retard the growth of tumor cells.
出处
《山东大学学报（医学版）》
CAS
2003年第1期4-6,共3页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金资助项目(30070341
39970333)
关键词
乙型肝炎病毒
反义寡核苷酸类
肝细胞癌
Hepatitis B virus
Oligonucleotides,antisense
Carcinoma,hepatocyte
