tRNA-包埋核酶在HepG2细胞中的抗HBV活性

Anti HBV Activity of tRNA embedded Ribozymes in HepG2 Cells

已知 ｔ Ｒ Ｎ Ａ包埋核酶比裸露核酶在胎牛血清和 Ｈｅｐ Ｇ２ 细胞抽提液中有较高的稳定性 构建了 ｈ Ｃ Ｍ Ｖ 启动子驱动下的抗 Ｈ Ｂ Ｖ（ａｄｒ 亚型）的 ｔ Ｒ Ｎ Ａ包埋核酶基因质粒，与携带 Ｈ Ｂ Ｖ 基因的ｐ１２Ⅱ质粒共转染 Ｈｅｐ Ｇ２ 细胞，用 Ｇ４１８ 筛选抗性细胞 分析稳定表达细胞中的 Ｈ Ｂ Ｖ Ｒ Ｎ Ａ， Ｈ Ｂ Ｖ 抗原合成和新生 Ｄ Ｎ Ａ 合成，表明ｔ Ｒ Ｎ Ａ包埋核酶比裸露核酶有较高的抑制 Ｈ Ｂ Ｖ 活性．ｔ Ｒ Ｎ Ａ包埋核酶和裸露核酶分别使靶 Ｒ Ｎ Ａ 减少 ８２％ ～８７％ 和 ７５％ ～８１％ ，抗原减少 ７３％ ～８０％ 和７０％ ～７４％ 以及新生 Ｄ Ｎ Ａ 减少 ７４％ ～７６％ 和 ６７％ ～７１％ 结果指出，核酶，特别是 ｔ Ｒ Ｎ Ａ包埋核酶，对 Ｈｅｐ Ｇ２ 细胞中 Ｈ Ｂ Ｖ 表达和复制有明显抑制作用，可能作为 Ｈ Ｂ Ｖ 基因治疗的手段之一

As is known,the stability of tRNA embedded ribozymes in the fetal bovine serum or cytoplasmic extracts of HepG2 cells is higher than that of the naked ribozymes In comparison with the anti HBV activities of the two kinds of ribozymes,the plasmids containing ribozyme genes under the control of hCMV promoter with upstream TK promoter/Neo gene were constructed and co transfected in HepG2 cells with a plasmid p1 2 Ⅱ carrying genome of HBV respectively Then the cells were selected with G418 The HBV RNAs,HBV antigens and progeny DNAs of G418 resistant and stably expressed cells were determined The results indicated that the anti HBV activity of tRNA embedded ribozymes was higher than that of naked ribozymes The tRNA embedded ribozymes and naked ribozymes were able to destroy the target RNA at 82%～87% and 75%～81%,to decrease the antigens at 73%～80% and 70%～74% as well as to inhibit the synthesis of progeny DNA at 74%～76% and 67%～71% respectively The data suggest that the ribozyme,especially tRNA embedded ribozyme,may be an important addition to the therapeutic modalities for the treatment of HBV infection