HBV preS2起始区反义核酸对人肝癌细胞在裸鼠体内生长的抑制作用

Inhibition of in vivo growth of gepatoma cells by antisense phosphorothioate oligodeoxynucleotides complementary to the initiator of HBV pre-S2

PCR合成互补于HBVpre S2起始区的反义寡核苷酸 (as preS2 ) ,以HBVDNA转染的HepG2 2 15细胞接种裸鼠制备人肝癌模型 ,通过连续用药、一次性用药、混合用药等三种途径研究其对人肝癌裸鼠模型的抑瘤生长及体内抗HBV作用。流式细胞术 (FCM)分析asON诱导瘤细胞凋亡作用 ,ELLSA法检测反义核酸作用裸鼠血清中HBV抗原含量的变化。结果显示 ,一次性与混合用药组裸鼠均表现为成瘤潜伏期延长 ,成瘤率明显低于瘤细胞对照未用药组 (P <0 0 5 ) ,瘤体生长缓慢。裸鼠瘤内连续用药 ,在 48hFCM测凋亡峰值为 39 5 7%,S期细胞降低显著 ,生理盐水与无关序列对照分别为 7 92 %、10 89%,提示as preS2可能诱导S期细胞凋亡。as preS2对荷瘤鼠HBeAg和HB sAg的抑制率分别为 45 %和 6 5 %。提示 ,as preS2可有效抑制体内HBV抗原表达 ,对人肝癌裸鼠在体内的生长有明显的抑制作用。

To Study the inhibitory effect of antisense phosphorothioate oligodeoxynucleotides(asON) complementary to the initiator of HBV pre-S2(as-preS2) on the growth of hepatoma cells and anti-HBV effect in vivo.BALB/c(nu/nu) mice were injected with HepG2.2.15 cells and then were divided into three groups. Group 1, once tumors were established, these animals were respectively given as-preS2 and saline, control sequence. Apoptosis was observed in animals given 100μg/day as-preS2, but not in saline or control sequence -treated animals. The apoptosis peaks were 39.57%, 39.18% respectively, at the time day 2 and day 4 after as-preS2 treatment. Group 2, animals were given 2.2.15cells and 100μg/mouse as-preS2 together. Group 3, animals were given 100μg/mouse after 24h given 2.2.15cells. In the second and third groups, a prolonged period of tumorigenesis was observed, but not following in control animals. The anti-HBV effect were assayed with ELISA. The inhibition rate on HBsAg and HBeAg was 65% and 45% respectively. These results demonstrated the anti-tumour activity of as-preS2 in vivo and the potential utility of short oligonucleotides targeted to preS2 as tumour cell inhibitors.