Apoptin enhances the oncolytic activity of vaccinia virus in vitro

• Published in: Molecular biology (New York) Vol. 47; no. 5; pp. 733 - 742
• Main Authors: Kochneva, G. V., Babkina, I. N., Lupan, T. A., Grazhdantseva, A. A., Yudin, P. V., Sivolobova, G. F., Shvalov, A. N., Popov, E. G., Babkin, I. V., Netesov, S. V., Chumakov, P. M.
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01-09-2013; Springer Nature B.V
• Subjects: Antitumor activity; Biochemistry; Biomedical and Life Sciences; Cancer; Cell culture; Cell Molecular Biology; Gene expression; Genomes; Growth factors; Human Genetics; Insertion; Life Sciences; Molecular biology; Oncolysis; Proteins; Puromycin; Tumor cell lines; Virulence; Viruses; recombinant vaccinia virus; puromycin; apoptin; virus growth factor; transient dominant selection; oncolytic activity
• ISSN: 0026-8933; 1608-3245
• DOI: 10.1134/S0026893313050075

The chicken anemia virus gene that encodes apoptin, a selective killer of cancer cells, was synthe-sized and inserted into the vaccinia virus (strain L-IVP) genome. The insertion replaces a major part of the viral
C11R
gene that encodes viral growth factor, which is important for virulence. The recombinant virus VVdGF-ApoS24/2 was obtained by transient dominant selection using the gene of puromycin resistance as a selective marker. The expression of the apoptin gene from a synthetic early-late promoter of vaccinia virus ensured the efficient accumulation of the target protein in VVdGF-ApoS24/2-infected cells. Although recombinant apoptin carried the signal peptide of the virus growth factor at the N-end, the protein was not secreted into the culture medium. The recombinant virus VVdGF-ApoS24/2 exhibited significantly higher selective lytic activity in human cancer cell lines (A549, A431, U87MG, RD, and MCF7) than the parent strain L-IVP and its VVdGF2/6 variant with
C11R
deletion. These results suggest that the use of apoptin can be an efficient means of enhancing the natural anticancer activity of vaccinia virus.