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A novel HBV antisense RNA gene delivery system targeting hepatocellular carcinoma
Chun-Hong Ma1, Wen-Sheng Sun, Pei-Kun Tian
1Institute of Immunology,Medical College of Shandong University, Jinan 250012, Shandong Province, China.
Aim:
To construct a novel HBV antisense RNA delivery system targeting hapatocellular carcinoma and study its inhibitory effect in vitro and in vivo.
Methods:
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.
Results:
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).
Conclusion:
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.
Insights
A novel delivery system effectively inhibits Hepatitis B virus (HBV) and hepatocellular carcinoma. This system targets liver tumors, reducing HBV antigens and tumor size in preclinical studies.
Area of Science:
- Hepatocellular Carcinoma Research
- Antisense RNA Therapeutics
- Viral Hepatitis Treatment
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern, often associated with chronic Hepatitis B virus (HBV) infection.
- Developing targeted therapies to inhibit both HBV replication and HCC progression is crucial.
Purpose of the Study:
- To construct and evaluate a novel HBV antisense RNA delivery system for targeting hepatocellular carcinoma.
- To investigate the in vitro and in vivo inhibitory effects of this system on HBV and HCC.
Main Methods:
- A novel HBV antisense RNA delivery system, the AFP-enhancing 4-element complex, was constructed using GE7 peptide, HA20 protein, and a hepatocarcinoma-specific HBV antisense expression vector.
- In vitro studies utilized HepG2.2.15 cells to assess the complex's inhibition of HBV antigen expression (HBsAg, HBeAg) via ELISA.
- In vivo studies involved BALB/c nude mice bearing HepG2.2.15 tumors, examining HBV antisense RNA expression by RT-PCR and measuring tumor size following complex administration.
Main Results:
- The AFP-enhancing 4-element complex efficiently trapped DNA, indicating successful system construction.
- Transfection with the complex significantly reduced HBsAg (33.4%) and HBeAg (58.5%) expression in HepG2.2.15 cells.
- In vivo, HBV antisense RNA was specifically expressed in liver tumor cells, and tumor size was significantly reduced (0.995 cm vs. 2.215 cm) after treatment.
Conclusions:
- The AFP-enhancing 4-element complex serves as an effective delivery system for HBV antisense RNA targeting hepatocellular carcinoma.
- This system demonstrates significant potential for inhibiting both HBV replication and liver tumor growth in vitro and in vivo.