A novel HBV antisense RNA gene delivery system targeting hepatocellular carcinoma

• Published in: World journal of gastroenterology : WJG Vol. 9; no. 3; pp. 463 - 467
• Main Authors: Ma, Chun-Hong, Sun, Wen-Sheng, Tian, Pei-Kun, Gao, Li-Fen, Liu, Su-Xia, Wang, Xiao-Yan, Zhang, Li-Ning, Cao, Ying-Lin, Han, Li-Hui, Liang, Xiao-Hong
• Format: Journal Article
• Language: English
• Published: United States Institute of Immunology, Medical College of Shandong University, Jinan 250012, Shandong Province, China%National Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai 200032, China 01.03.2003; Baishideng Publishing Group Inc
• Subjects: Animals; Carcinoma, Hepatocellular - genetics; Gene Transfer Techniques; Hepatitis B virus - genetics; Liver Cancer; Liver Neoplasms - genetics; Male; Mice; Mice, Inbred BALB C; RNA, Antisense; RNA, Viral - genetics; Tumor Cells, Cultured
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v9.i3.463

To construct a novel HBV antisense RNA delivery system targeting hapatocellular carcinoma and study its inhibitory effect in vitro and in vivo. GE7,a 16-peptide specific to EGFR, and HA20,a homologue of N-terminus of haemagglutinin of influenza viral envelope protein, were synthesized and conjugated with polylysin. The above conjugates were organized into the pEBAF-as-preS2, a hepatocarcinoma specific HBV antisense expression vector, to construct a novel HBV antisense RNA delivery system, named AFP-enhancing 4-element complex. Hepatocelluar carcinoma HepG2.2.15 cells was used to assay the in vitro inhibition of the complex on HBV. Expression of HBV antigen was assayed by ELISA. BALB/c nude mice bearing HepG2.2.15 cells were injected with AFP-enhancing 4-element complex. The expression of HBV antisense RNA was examined by RT-PCR and the size of tumor in nude mice were measured. The AFP-enhancing 4-element complex was constructed and DNA was completely trapped at the slot with no DNA migration when the ratio of polypeptide to plasmid was 1:1. The expression of HBsAg and HBeAg of HepG2.2.15 cells was greatly decreased after being transfected by AFP-enhancing 4-element complex. The inhibitory rates were 33.4 % and 58.5 % respectively. RT-PCR showed HBV antisense RNA expressed specifically in liver tumor cells of tumor-bearing nude mice. After 4 injections of AFP-enhancing 4-element complex containing 0.2 micro g DNA, the diameter of the tumor was 0.995 cm+/-0.35, which was significantly smaller than that of the control groups(2.215 cm+/-0.25, P<0.05). AFP-enhancing 4-element complex could deliver HBV antisense RNA targeting on hepatocarcinoma and inhibit both HBV and liver tumor cells in vitro and in vivo.