Abstract
Objective
The aim of the study was to observe the transfection efficacy of hepatitis B virus envelope (HBVE) and evaluate its ability as a gene transfer vector for liver cancer cells.
Methods
To obtain HBVE, the supernatant fluid of HepG 2.2.15 cells was mixed with a PEG8000 solution for concentration and was inactivated by β-propiolactone. The acquired HBVE was used to pack pIRES2-EGFP to test its package ability. Then, we examined its quantity and quality with ELISA, PCR, SDS-PAGE and electron microscopy. The pIRES2-EGFP was packed with HBVE and obtained the product HBVE-GFP. The pIRES2-EGFP was packed with liposome and obtained the product liposome-GFP. HBVE-GFP and liposome-GFP were used to transfer HepG 2 cells to study the transfection efficiency. HBVE-GFP was used to transfer HepG 2, A549, HeLa and FB cells to study the targeting ability. The green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometry.
Results
1. The acquired HBVE could retain the surface protein HBsAg + pre S1 + pre S2 and had no virus DNA. It had good package ability for pIRES2-EGFP. 2. Transfection efficiency: The GFP could be observed in both the liposome group and HBVE group under the fluorescent microscope. But the HBVE group had a higher fluorescent intensity than liposome group. The transfection rate of liposome group was 49.97% ± 2.37% while the HBVE group was 70.65% ± 3.15% and the fluorescent intensity of the HBVE group was 3–4 times (P = 0.000) for liposome group with the determination of flow cytometry. 3. Targeting ability: The GFP could be observed in the four groups under the fluorescent microscope. The HepG 2 group had the highest fluorescent intensity among the four groups. The transfection rate of HepG 2 group was 71.35% ± 0.03% which was highly expressed than other groups (P = 0.000) and the fluorescent intensity of the HepG 2 group was 2–3 times (P = 0.000) for the other 3 groups with the determination of flow cytometry.
Conclusion
HBVE can be constructed successfully with the methods of PEG8000 and β-propiolactone from the supernatant fluid of HepG 2.2.15 cells. The HBVE can be a candidate gene transfer vector for liver cancer cells.
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Supported by a grant from the National Natural Sciences Foundation of China (No. 30100189).
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Pan, D., Wang, W., Wang, D. et al. Hepatitis B virus envelope as a targeting gene transfer vector for hepatic cancer cells. Chin. -Ger. J. Clin. Oncol. 8, 447–452 (2009). https://doi.org/10.1007/s10330-009-0084-9
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DOI: https://doi.org/10.1007/s10330-009-0084-9