Expression and cytotoxic effect of recombinant Newcastle Disease Virus (rNDV) vector expressing enhanced green fluorescent gene in JHH5 cell line

Authors

  • Lubna Muhi Rasoul
  • Risala H. Allami
  • Ammar Luay Alshibib
  • Bahaa Abdullah laftaah Al-Rubaii
  • Tahreer Hadi Sale

DOI:

https://doi.org/10.51248/.v43i1.2425

Keywords:

Newcastle disease virus, rNDV, GFP gene, MTT assay, JHH5 cell line

Abstract

Introduction and Aim: In medicine, the term ‘cancer’ is used to describe a group of disorders defined by uncontrolled cell growth and the ability to invade and metastasize to other organs. As a result of the body's ability to fight conventional cancer treatments, it is essential to look for alternative therapeutic avenues. Recent years have seen a rise in the application of reverse genetics techniques including the Newcastle disease virus as an oncolytic agent in tumor models of immunocompetent malignancy. In preclinical studies, recombinant NDV (rNDV-GFP) that expresses foreign genes was shown to be more effective in cancer therapy. The goal of this study was to examine the in vitro anticancer effects of GFP-expressing, genetically modified Newcastle disease virus (rClone3-GFP) strains on the JHH5 hepatocellular carcinoma cell line.

 

Materials and Methods: To generate a recombinant NDV, the GFP gene was inserted into the genome of an NDV strain (NDVClone30) by reverse genetics technology, where it now resides in the region of the genome occupied by the F and HN genes (rNDV-GFP). Using the MTT assay, we evaluated rNDV-GFP for its oncolytic potential against JHH5 cancer cells. Cytopathic effects were analyzed using fluorescent and light microscopy.

 

Results: We observed that 96 hours post infection, the recombinant virus was capable of inducing tumor cell death in a time-dependent way.

 

Conclusion: Recombinant NDV strains expressing GFP, demonstrated good results in inhibiting tumor growth. Our results pave the door for the application of recombinant NDV as a viral vector for the treatment of liver cancer.

Author Biographies

Lubna Muhi Rasoul

Department of Biology, University of Baghdad, Baghdad, Iraq

Risala H. Allami

Department of Molecular and Medical Biotechnology, College of Biotechnology, Al-Nahrain University, Jadriya, Baghdad, Iraq

Ammar Luay Alshibib

Al-Farahidi University, College of Dentistry, Baghdad, Iraq

Bahaa Abdullah laftaah Al-Rubaii

Department of Biology, University of Baghdad, Baghdad, Iraq

Tahreer Hadi Sale

Department of Biology, College of Science, Mustansiriyah University, Baghdad, Iraq

References

Torre, L.A., Siegel, R. L., Ward, E.M., Jemal, A. Global Cancer Incidence and Mortality Rates and Trends—An Update Global Cancer Rates and Trends—An Update. Cancer epidemiology, biomarkers & prevention.2016; 25(1):16-27.

Ferguson, M.S., Lemoine, N.R., Wang, Y. Systemic delivery of oncolytic viruses: hopes and hurdles. Advances in Virology. 2012; 1-14.

Lichty, B.D., Breitbach, C.J., Stojdl, D.F., Bell, J.C. Going viral with cancer immunotherapy. Nature Reviews. Cancer. 2014; 14(8):559-567.

Naomi, D.S., Kirn, A. H., David, H., Bell, J. C., Breitbach, C.J. Double trouble for tumours: Exploiting the tumour microenvironment to enhance anticancer effect of oncolytic viruses. Cytokine & Growth Factor Reviews. Recent Advances in the Development of Oncolytic Viruses as Cancer Therapeutics. 2010; 21(2): 135-141.

Alemany, R. Viruses in cancer treatment. Clinical and Translational Oncology. 2013; 153):182-188.

Roberts, M.S., Lorence, R. M., Groene, W. S., Bamat, M.K. Naturally oncolytic viruses. Current Opinion in Molecular Therapeutics. 2006; 8(4):314-321.

Phillips, G.J., Green fluorescent protein--a bright idea for the study of bacterial protein localization. FEMS Microbiology Letters. 2001; 204(1): 9-18.

Hoffman, R.M. Application of GFP imaging in cancer. Lab Invest. 2015; 95(4):432-452.

Forner, A., Llovet, J.M., Bruix, J. Hepatocellular carcinoma. The Lancet. 2012;379 (9822):1245-1255.

‘Global Cancer Observatory’. gco.iarc.fr. Retrieved 2021-06-24.

Rumgay, H., Arnold, M., Ferlay, J., Lesi, O., Cabasag, C.J., Vignat, J., et al., Global burden of primary liver cancer in 2020 and predictions to 2040. J Hepatol. 2022; 77(6):1598-1606.

Ali, S.M., Laftah, B.A., Al-Shammary, A.M., Salih, H.S. Study the role of bacterial neuraminidase against adenocarcinoma cells in vivo. InAIP Conference Proceedings. 2021; 2372: 030009.

Rasoul, L. M., Marhoon, A.A., Albaayit, S.F.A., Ali, R.W., Saleh, T. H., Al-Rubaii, B.A.L. Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system. Biomedicine. 2022; 42(5):938-942.

Ali, S.M., Lafta, B.A., Al-Shammary, A.M., Salih, H.S. In vivo oncolytic activity of non-virulent Newcastle disease virus Iraqi strain against mouse mammary adenocarcinoma. AIP Conference Proceedings. 2021; 2372: 030010.

Abdulrazaq, R.A., Mahmood, W.S., Alwan, B., Saleh, T.H., Hashim, S.T., Al-Rubaii, B.A.L. Biological Study of protease produced by clinical isolates of Staphylococcus aureus. Research Journal of Pharmacy and Technology.2022; 15(12):5415-5420.

Al-Rubii, B.A.L. Cloning LasB gene of Pseudomonas aeruginosa elastase 10104-2aI in E. coli BL21 and E. coli DH5alpha and investigated their effect on the stripping of Vero cells. Pakistan J Biotechnol. 2017; 14(4):697-705.

Abdul-Gani, M., Laftaah, B.A. Purification and characterization of chondroitinase ABC from Proteus vulgaris, an Iraqi clinically isolated. Curr Sci., 2017; 113(11):2134-2140.

Rasoul, L.M., Marhoon, A.A., Albaayit, S.F.A., Ali, R.W., Saleh, T.H., Al-Rubaii, B.A.L. Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system. Biomedicine. 2022; 42(5):938-942.

Cui, Y., Qin, S., Jiang, P. Chloroplast transformation of Platymonas (Tetraselmis) subcordiformis with the bar Gene as Selectable Marker, PLoS One. 2014; 9(6): e98607.

He, J., Pan, Z., Tian, G., Liu, X., Liu, Y., Guo, X., et al., Newcastle disease virus chimeras expressing the Hemagglutinin- Neuraminidase protein of mesogenic strain exhibits an enhanced anti-hepatoma efficacy. Virus research. 2016; 221:23-29.

Pan, Z., He, J., Rasoul, L. M., Liu, Y., Che, R., Ding, Y., et al., Identification of optimal insertion site in recombinant Newcastle Disease Virus (rNDV) vector expressing foreign gene to enhance its anti-tumor effect. PloS one. 2016; 11(10): e0164723.

Walther, W., Stein, U.S. Newcastle Disease Virus: A promising vector for viral therapy, immunotherapy, and gene therapy of cancer. Gene Therapy of Cancer. 2009; 542: 565-605.

Pantua, H.D., Mcginners, L., Peeples, M.E, Morrison, T.G. Requirements for the assembly and release of Newcastle Disease Virus like particles. J. Virol. 2006; 80(22): 11062-11075.

Yang, M., Li, L., Jiang, P., Moossa, A.R., Penman, S., Hoffman, R.M., Dual-color fluorescence imaging distinguishes tumor cells from induced host angiogenic vessels and stromal cells. Proc Natl Acad Sci USA. 2003; 100: 14259-14262.

Elmore, S. Apoptosis: A review of programmed cell death. Toxicol Pathol. 2007; 35(4): 495-516.

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Published

2023-02-26

How to Cite

1.
Rasoul LM, H. Allami R, Alshibib AL, Al-Rubaii BA laftaah, Hadi Sale T. Expression and cytotoxic effect of recombinant Newcastle Disease Virus (rNDV) vector expressing enhanced green fluorescent gene in JHH5 cell line. Biomedicine [Internet]. 2023 Feb. 26 [cited 2024 Mar. 3];43(1):205-9. Available from: https://biomedicineonline.org/index.php/home/article/view/2425

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