Producing Vesicular Stomatitis Virus G (VSVG) Protein and Assessment of its Cytotoxic Activity against Breast Cancer Cells

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Biology, School of Sciences, Islamic Azad University, Tehran Sciences and Research Branch, Tehran, Iran

2 Assistant Professor, Department of Biotechnology, School of New Sciences and Technology, University of Isfahan, Isfahan, Iran

3 Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 Assistant Professor, Department of Biology, School of Sciences, Islamic Azad University, Tehran Sciences and Research Branch, Tehran, Iran

Abstract

Background: The vesicular stomatitis virus G (VSVG) protein is a transmembran glycoprotein, which is involved in virus attachment to the specific receptor at the cell surface. This protein has been widely used to study therapeutic gene delivery. However, the major limitation of its usage in gene therapy is toxicity of protein for host cells in high concentrations.Methods: The vesicular stomatitis virus G protein was produced via transfection of HEK-293T cells with using polyfection kit. The cytotoxic activity was examined against MCF-7 and MDA-MB-231 breast cancer cell lines and human embryonic kidney normal cell line (HEK293) using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and morphology changes assay.Findings: The cytotoxic activity of vesicular stomatitis virus G protein against MCF-7 and MDA-MB-231 cells was significantly more than that of the normal cells (P < 0.05). In addition, cancer cells treated with vesicular stomatitis virus G protein showed morphology changes.Conclusion: The results indicated that cytotoxic activity of vesicular stomatitis virus G protein against MCF-7 and MDA-MB-231 cells was more than that of the normal cells and it could be an appropriate candidate for in-vivo testing as cytotoxic agent.

Keywords

References

  1. Sanders DA. No false start for novel pseudotyped vectors. Curr Opin Biotechnol 2002; 13(5): 437-42.
  2. Beyer WR, Westphal M, Ostertag W, von LD. Oncoretrovirus and lentivirus vectors pseudotyped with lymphocytic choriomeningitis virus glycoprotein: generation, concentration, and broad host range. J Virol 2002; 76(3): 1488-95.
  3. Galipeau J, Li H, Paquin A, Sicilia F, Karpati G, Nalbantoglu J. Vesicular stomatitis virus G pseudotyped retrovector mediates effective in vivo suicide gene delivery in experimental brain cancer. Cancer Res 1999; 59(10): 2384-94.
  4. Rose JK, Whitt MA. Rhabdoviridae: The viruses and their replication. In: Knipe DM, Howley PM. Field's Virology. 4th ed. Philadelphia, PA: Lippincott Williams and Wilkins; 2001. p. 1221–44.
  5. Gaudin Y, Ruigrok RW, Knossow M, Flamand A. Low-pH conformational changes of rabies virus glycoprotein and their role in membrane fusion. J Virol 1993; 67(3): 1365-72.
  6. Yu H, Eton D, Wang Y, Kumar SR, Tang L, Terramani TT, et al. High efficiency in vitro gene transfer into vascular tissues using a pseudotyped retroviral vector without pseudotransduction. Gene Ther 1999; 6(11): 1876-83.
  7. Miyanohara A. Preparation of vesicular stomatitis virus-G (VSV-G) conjugate and its use in gene transfer. Cold Spring Harb Protoc 2012; 2012(4): 453-6.
  8. Qiao J, Moreno J, Forshaw M, Diaz RM, Vile RG. 741. VSV-G pseudotyped, MLV-based, semi-replication-competent retroviruses for cancer gene therapy. Mol Ther 2004; 9(S1): S282.
  9. Amado RG, Chen IS. Lentiviral vectors--the promise of gene therapy within reach? Science 1999; 285(5428): 674-6.
  10. Coil DA, Miller AD. Phosphatidylserine is not the cell surface receptor for vesicular stomatitis virus. J Virol 2004; 78(20): 10920-6.
  11. Sun X, Belouzard S, Whittaker GR. Molecular architecture of the bipartite fusion loops of vesicular stomatitis virus glycoprotein G, a class III viral fusion protein. J Biol Chem 2008; 283(10): 6418-27.
  12. Dobrzynska I, Szachowicz-Petelska B, Sulkowski S, Figaszewski Z. Changes in electric charge and phospholipids composition in human colorectal cancer cells. Mol Cell Biochem 2005; 276(1-2): 113-9.
Journal of Isfahan Medical School
Volume 33, Issue 325 - Serial Number 325
January and February 2015
Pages 221-230

Files

History

  • Receive Date: 29 November 2014
  • Revise Date: 02 November 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics