Research of gene therapy for viral hepatitis using anti-sense DNA

亚洲糖蛋白受体介导的基因传递是肝细胞选择性基因传递的最有效方法。体外转染DNA到HepG2细胞，转染效率最高，载体-DNA比为480:1。当溶酶体酶抑制剂氯喹转染HepG2细胞时，添加50 mmol/L时，效果提高22倍。在大约3%的HepG2细胞中观察到转移的dna编码产物的表达。将载体-^<32> p标记的DNA复合物注入小鼠尾血管后，大部分^<32> p标记的DNA被肝脏吸收。转染^<35> s标记的DNA后，活体标本的显微放射自显影显示所传递的DNA位于肝实质细胞中。在体内测试基因传递的功效时，氯喹有效地提高了疗效。此外，氯喹与门静脉内注射联合使用可显著增强asialal糖蛋白受体介导的基因传递在体内的转染。因此，asialal糖蛋白受体介导的反义DNA基因转移是肝细胞的选择性基因传递，可能是病毒性肝炎基因治疗的有效方法。

Asialoglycoprotein receptor mediated gene delivery is the most useful method for selective gene delivery to the hepatocytes. In vitro transfection of DNA to HepG2 cells, the transfection efficacy was highest at the carrier-DNA ratio of 480 : 1. Whne a lysosomal enzyme inhibitor, chloroquine was used for transfection into HepG2 cells, a 22-fold increase of efficacy was observed with addition of 50 mmol/L.The expression of transfered DNA-encoded products was observed in approximately 3% of HepG2 cells. When the carrier-^<32>P-labeled DNA complex was injected into the tail vessel of the mouse, most ^<32>P-labeled DNA were taken up by the liver. Microautoradiography of the live specimens after transfection of ^<35>S-labeled DNA indicated that the delivered DNA was located in the liver parenchymal cells.When the efficacy of gene delivery was tested in vivo, chloroquine effectively increased the efficacy. Moreover, the combination of chloroquine and intraportal injection strongly enhanced the in vivo transfection with asialoglycoprotein receptor mediated gene delivery.Thus, asialoglycoprotein receptor mediated gene transfer of anti-sense DNA is the selective gene delivery to the hepatocytes and may be the useful method of gene therapy for viral hepatitis.