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目的定量观察双位点核酶对细胞内ＨＢＶ基因表达的抑制作用。方法构建针对ＨＢＶＣ区双位点核酶的真核表达载体（ｐＢＢＳ２１２－Ｒｚ），将该质粒与ｐｌ．２Ⅱ（含ＨＢＶ全序列），共转染人肝细胞癌（ＨＨＣＣ）细胞株，对转染细胞所表达ＨＢｅＡｇ进行间接免疫荧光染色，用共聚焦显微镜对其含量进行定量分析。结果共转染细胞可以表达ＨＢｅＡｇ，同对照组相比，核酶组荧光量明显下降。结论在核酶与ＨＢＶ共转染细胞中，核酶通过对ＨＢＶｍＲＮＡ的剪切作用，使ＨＢｅＡｇ的表达量明显受到抑制。
Objective To quantitatively observe the inhibitory effect of double-site ribozyme on HBV intracellular HBV gene expression. Methods The eukaryotic expression vector (pBBS212-Rz) targeting the double-site ribozyme of HBVC region was constructed. 2Ⅱ (including full-length HBV) were transfected into human hepatocellular carcinoma (HHCC) cell lines. The expression of HBeAg in transfected cells was detected by indirect immunofluorescence staining. The content of HBeAg was quantitatively analyzed by confocal microscopy. Results The co-transfected cells could express HBeAg. Compared with the control group, the amount of fluorescence in the ribozyme group decreased significantly. Conclusion In the co-transfected cells with ribozyme and HBV, the ribozyme can significantly inhibit the expression of HBeAg through the shearing of HBV mRNA.