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AIM TO construct a novel HBV antisense RNA deliverysystem targeting hapatocellular carcinoma and study itsinhibitory effect in vitro and in vivo.METHODS:GE7,a 16-peptide specific to EGFR,and HA20,a homologue of N-terminus of haemagglutinin of influenzaviral envelope protein,were synthesized and conjugatedwith polylysin.The above conjugates were organized intothe pEBAF-as-preS2,a hepatocarcinoma specific HBVantisense expression vector,to construct a novel HBVantisense RNA delivery system,named AFP-enhancing 4-element complex.Hepatocelluar carcinoma HepG2.2.15 cellswas used to assay the in vitro inhibition of the complex onHBV.Expression of HBV antigen was assayed by ELISA.BALB/c nude mice bearing HepG2.2.15 cells were injectedwith AFP-enhancing 4-element complex.The expression ofHBV antisense RNA was examined by RT-PCR and the sizeof tumor in nude mice were measured.RESULTS:The AFP-enhancing 4-element complex wasconstructed and DNA was completely trapped at the slotwith no DNA migration when the ratio of polypeptide toplasmid was t:l.The expression of HBsAg and HBeAg ofHepG2.2.15 cells was greatly decreased after beingtransfected by AFP-enhancing 4-element complex.Theinhibitory rates were 33.4 % and 58.5 % respectively.RT-PCR showed HBV antisense RNA expressed specifically inliver tumor cells of tumor-bearing nude mice.After 4injections of AFP-enhancing 4-elernent complex containing0.2 μg 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).CONCLUSION:AFP-enhancing 4-element complex coulddeliver HBV antisense RNA targeting on hepatocarcinoma andinhibit both HBV and liver tumor cells in vitro and in vivo.
AIM TO construct a novel HBV antisense RNA deliverysystem targeting hapatocellular carcinoma and study its inhibitory effect in vitro and in vivo. METHODS: GE7, a16-peptide specific to EGFR, and HA20, a homologue of N-terminus of haemagglutinin of influenzaviral envelope protein, were synthesized and conjugated with polylysin. Above conjugates were organized intothe pEBAF-as-preS2, a hepatocarcinoma specific HBVantisense expression vector, to construct a novel HBVantisense RNA delivery system, named AFP-enhancing 4-element complex. Hepatocellular carcinoma HepG2.2.15 cellswas 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 .RESULTS: The AFP-enhancing 4-element complex wasconstructed and DNAwas completely trapped at the slotwith no DNA migration when the ratio of polypeptide toplasmid was t: l. The expression of HBsAg and HBeAg ofHepG2.2.15 cells was greatly decreased after beingtransfected by AFP-enhancing 4-element complex .hibitory rates were 33.4% and 58.5% respectively. RT-PCR showed HBV antisense RNA expressed specifically inliver tumor cells of tumor-bearing nude mice. After 4 injections of AFP-enhancing 4-elernent complex containing 0.2 μg DNA, the diameter of the tumor was 0.995 cm ± 0.35, which was significantly smaller than that of the control (2.215 cm ± 0.25, P <0.05) .CONCLUSION: AFP-enhancing 4-element complex coulddeliver HBV antisense RNA targeting on hepatocarcinoma and inhibits both HBV and liver tumor cells in vitro and in vivo.