核酶对人HCC细胞内HBeAg的抑制作用

Inhibitory effect of ribozyme on HBeAg in human HCC cells *

目的观察针对ＨＢＶＣ区双位点核酶对ＨＢｅＡｇ在细胞内表达的抑制作用．方法利用亚克隆技术，从质粒ｐＧＥＭＲｚ１２３切下ＥｃｏＲⅠＢａｍＨⅠ片段，双粘端克隆于真核表达质粒ｐＢＢＳ２１２中，将该质粒与ｐ１．２Ⅱ（含ＨＢＶ全序列），共转染人ＨＣＣ细胞（人肝癌细胞株），观察该核酶在细胞内对ＨＢｅＡｇ合成的抑制作用．结果在人ＨＣＣ细胞中ｐ１．２Ⅱ可表达出ＨＢｅＡｇ；该核酶对ＨＢｅＡｇ的合成抑制率达６５％．结论该双位点核酶可能通过针对ＨＢＶＣ区基因的剪切作用，阻断Ｃ区基因的表达。

AIM To observe the inhibition of HBeAg expression in the human hepatic carcinoma cell (HCC) transfected by HBV genome with two unit ribozyme against HBV. METHODS By using subclone technique, the two unit ribozyme gene cut from pGEM Rz 123 was ligated into the eukaryotic expression vector pBBS212. The recombinant plasmid pBBS212 Rz and p1.2Ⅱ plasmid carrying genome of adr subtype HBV were cotransfected into the human HCC cell using lipofectamine. The endocellular e antigen of HBV was detected by ELISA. RESULTS The cotransfection cell did express HBV e antigen, and two unit ribozyme lowered the level of HBeAg expressed in coinfected cells by up to 65%. CONCLUSION The two unit ribozyme can inhibit HBV expression in cotransfected cell.