Generation of a liver targeting fusion interferon and its bioactivity analysis in vitro.
The objective of this study was to generate a liver targeting fusion interferon, galactosyl-human serum albumin-interferon alpha2b (G-HSA-IFN) and to evaluate its bioactivity in vitro on HepG2.2.15 cells which express hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg). The cell proliferation was determined by Sulpho Rhodamine B (SRB) staining method and flow cytometry (FCM) assay. Hochest33342 and Propidium Iodide (PI) double staining and Western blot analysis of Bcl-2/Bax were also performed to evaluate cell lethality and apoptosis. The concentrations of HBsAg and HBeAg secreted in culture supernatant were detected using Enzyme-Linked Immunosorbent Assay (ELISA). The results demonstrated that G-HSA-IFN could inhibit the proliferation of HepG2.2.15 cells and the cell cycle was arrested at G0/G1 phase. Western blotting results showed that the expression of Bcl-2 was inhibited in a dose-dependent manner while the expression of Bax was enhanced. The expression of HBsAg was inhibited by G-HSA-IFN in a dose-dependent manner, while no significant inhibiting effect on the expression of HBeAg was observed. Conclusively, G-HSA-IFN could not only significantly inhibit the HBsAg expression and the proliferation of HepG2.2.15 cells, but also induce the apoptosis of the target cells, rendering it a promising drug candidate for hepatitis B.
Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, China.
Gangming
G
Cai
Mengjun
M
Jiang
Yaoyuan
Y
Zhou
Xiaobo
X
Gu
Bo
B
Zhang
Meifen
M
Zou
Xingqin
X
Zhou
Jiandong
J
Bao
Guoxian
G
Cao
Rongjun
R
Zhang
D000998:Antiviral Agents / Q000008:administration & dosage* / Q000493:pharmacokinetics* / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D016503:Drug Delivery Systems; D004797:Enzyme-Linked Immunosorbent Assay; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000096:biosynthesis; D006513:Hepatitis B e Antigens / Q000032:analysis / Q000096:biosynthesis; D006801:Humans; D016898:Interferon-alpha / Q000008:administration & dosage* / Q000493:pharmacokinetics* / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016196:S Phase / Q000187:drug effects
D000998:Antiviral Agents; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D016898:Interferon-alpha