Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system

Authors

  • Lubna Muhi Rasoul
  • Abeer Ali Marhoon
  • Shaymaa Fadhel Abbas Albaayit
  • Rusul Waleed Ali
  • Tahreer Hadi Saleh
  • Bahaa Abdullah Laftah Al-Rubaii

DOI:

https://doi.org/10.51248/.v42i5.1934

Keywords:

rNDV, GPF gene, reverse genetic techniques, MTT, NCI-H727

Abstract

Introduction and Aim: Cancers are a complex group of genetic illnesses that develop through multistep, mutagenic processes which can invade or spread throughout the body. Recent advances in cancer treatment involve oncolytic viruses to infect and destroy cancer cells. The Newcastle disease virus (NDV), an oncolytic virus has shown to have anti-cancer effects either directly by lysing cancer cells or indirectly by activating the immune system. The green fluorescent protein (GFP) has been widely used in studying the anti-tumor activity of oncolytic viruses. This study aimed to study the anticancer effect of a recombinant rNDV-GFP clone on NCI-H727 lung carcinoma cell line in vitro.

 

Materials and Methods: The GFP gene was inserted to a NDV strain to create a recombinant NDV (rNDV- GFP) using reverse genetics technology. The MTT assay was used in evaluating the oncolytic effect of rNDV- GFP on the lung carcinoma NCI-H727 cells. Light and fluorescent microscopy was used to study the cytopathic effects of rNDV-GFP.

 

Results: MTT assay showed that rNDV-GPF inhibited the NCI-H727 tumor cell death in a time-dependent manner. A significant inhibitory effect (78.3%) for rNDV-GPF on cancer cells was observed at 96h in comparison to rNDV (22.7%) and the cytotoxicity rate was directly proportional to the MOI used. Microscopic studies showed rNDV-GPF to induce cytopathic effect post 24 h of infection.

 

Conclusion: The GFP-expressing recombinant NDV strains exhibited encouraging results in terms of tumor growth inhibition. Our research set the groundwork for employing recombinant NDV as an anticancer viral vector.

 

Author Biographies

Lubna Muhi Rasoul

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

Abeer Ali Marhoon

Al-Rafidain University College, Baghdad, Iraq

Shaymaa Fadhel Abbas Albaayit

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

Rusul Waleed Ali

Department of Medical Laboratory Technique, Dijlah University College, Baghdad, Iraq

Tahreer Hadi Saleh

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

Bahaa Abdullah Laftah Al-Rubaii

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

References

Albaayit SFA, Khan MA, Abdullah R. Zerumbone induces growth inhibition of Burkitt’s lymphoma cell line via apoptosis. Nat Volatiles Essent Oils. 2021;8(3): 56-63.

Albaayit SFA, Maharjan R, Khan M. Evaluation of hemolysis activity of Zerumbone on RBCs and brine shrimp toxicity. Baghdad Sci J. 2021;18(1): 65-69. DOI: https://doi.org/10.21123/bsj.2021.18.1.0065

Albaayit SFA, Maharjan R, Abdullah R, Noor MHM. Anti-Enterococcus faecalis, cytotoxicity, phytotoxicity, and anticancer studies on Clausena excavata Burum. f. (Rutaceae) Leaves. Biomed Res Int. 2021, 3123476 DOI: https://doi.org/10.1155/2021/3123476

Alemany, R., Viruses in cancer treatment. Clin Transl Oncol. 2013;15 (3):182-188. DOI: https://doi.org/10.1007/s12094-012-0951-7

Ferguson, M.S., Lemoine, N.R., Wang, Y. Systemic delivery of oncolytic viruses: hopes and hurdles. Advances in Virology. 2012; Special issue: 1-14. DOI: https://doi.org/10.1155/2012/805629

Rasoul, L.M., Ali, L.F., Mohammed, N.S. The promising anti-tumor impact of Newcastle Disease Virus expressing IL-2 and P53 genes in many cancer cell lines in vitro. Iraqi J. Biotechnol. 2019; 18(2):169-181.

Lichty, B.D., Breitbach, C.J., Stojdl, D.F., Bell, J.C., Going viral with cancer immunotherapy. Nat Rev Cancer. 2014; 14 (8):559-567. DOI: https://doi.org/10.1038/nrc3770

Wajid A, Wasim M, Rehmani SF, Bibi T, Ahmed N, Afonso CL. 2015. Complete genome sequence of a recent panzootic virulent Newcastle disease virus from Pakistan. Genome Announc 3(3):e00658-15. doi: 10.1128/genomeA.00658-15. DOI: https://doi.org/10.1128/genomeA.00658-15

Jinjiao He, Ziye Pan, Guiyou Tian, Xin Liu, Yunye Liu, Xiaochen Guo, et al., Newcastle disease virus chimeras expressing the hemagglutinin-neuraminidase protein of mesogenic strain exhibits an enhanced anti-hepatoma efficacy. 2016; 221:23-29. DOI: https://doi.org/10.1016/j.virusres.2016.04.023

Miller PJ, Decanini EL, Afonso CL. Newcastle disease: evolution of genotypes and the related diagnostic challenges. Infect Genet Evol 2010; 10:26–35. DOI: https://doi.org/10.1016/j.meegid.2009.09.012

Huang Z, Liu M, Huang Y. Oncolytic therapy and gene therapy for cancer: recent advances in antitumor effects of Newcastle disease virus. Discov Med. 2020;30(159):39-48.

Ziye Pan, Jinjiao He, Lubna M. Rasoul, Yunye Liu, Ruixiang Che, Yun Ding, et al., Identification of optimal insertion site in recombinant Newcastle Disease Virus (rNDV) vector expressing foreign gene to enhance its anti-tumor effect. 016, 221, pp. 23-29.

Rasoul, L.M., He, J., Khoso, M.H., Li, D. Use of viral vector to deliver IL-15 for cancer therapy: An overview. Indian Journal of Biochemistry & Biophysics. 2017; 54: 97-108.

Lam, H.Y., Yeap, S.K., Rasoli, M., Omar, A.R., Yusoff, K., Suraini, A.A. et al., Safety and clinical usage of Newcastle disease virus in cancer therapy. Biomed Research International. 2011; 2011: Article ID 718710. https://doi.org/10.1155/2011/718710. DOI: https://doi.org/10.1155/2011/718710

Vigil A, Park MS, Martinez O, Chua MA, Xiao S, Cros JF, Martínez-Sobrido L, Woo SL, García-Sastre A. Use of reverse genetics to enhance the oncolytic properties of Newcastle disease virus. Cancer Res. 2007;67(17):8285-8292. DOI: https://doi.org/10.1158/0008-5472.CAN-07-1025

Hoffman RM. Application of GFP imaging in cancer. Lab Invest. 2015;95(4):432-452. DOI: https://doi.org/10.1038/labinvest.2014.154

Yamamoto N, Yang M, Jiang P, Xu M, Tsuchiya H, Tomita K, et al., . Real-time imaging of individual fluorescent-protein color-coded metastatic colonies in vivo. Clin Exp Metastasis. 2003; 20(7):633-638. DOI: https://doi.org/10.1023/A:1027311230474

Albaayit, S.F.A., Khan, M.A., Abdullah, R., Mohd Noor, M.H. Ethyl acetate extract of Clausena excavata induces growth inhibition of non-small-lung cancer, NCI-H460, cell line via apoptosis. J Appl Biomed. 2021; 19(1):40-47. DOI: https://doi.org/10.32725/jab.2021.007

Heng, W.S., Kruyt, F.A., Cheah, S.C. Understanding lung carcinogenesis from a morphostatic perspective: Prevention and therapeutic potential of phytochemicals for targeting cancer stem cells. Int J Mol Sci. 2021; 22(11):5697. DOI: https://doi.org/10.3390/ijms22115697

Cui, Y., Qin, S. and Jiang, P. Chloroplast transformation of Platymonas (Tetraselmis) subcordiformis with the bar gene as selectable marker. PLOS One, 2014; 9(6): e98607. doi: 10.1371/journal.pone.0098607. DOI: https://doi.org/10.1371/journal.pone.0098607

Sung, H., Ferlay, J., Siegel, R.L., Laversanne, M., Soerjomataram, I., Jemal, A., Bray, F. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians. 2021;71 (3):209-249. DOI: https://doi.org/10.3322/caac.21660

Pantua, H.D., Mcginners, L.W., 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. DOI: https://doi.org/10.1128/JVI.00726-06

Zimmer, M., Green fluorescent protein (GFP): applications, structure and related photophysical behavior. Chem Rev. 2002; 102:759-781. DOI: https://doi.org/10.1021/cr010142r

Yang, M., Li, L., Jiang, P., Moossa, A.R., Penman, S., Hoffman, R.M., et al., Dual-color fluorescence imaging distinguishes tumor cells from induced host angiogenic vessels and stromal cells. Proc Natl Acad Sci USA. 2003;100: 14259-14262. DOI: https://doi.org/10.1073/pnas.2436101100

Susan, E. Apoptosis: A review of programmed cell death, Toxicol Pathol. 2007; 35(4): 495-516. DOI: https://doi.org/10.1080/01926230701320337

Yasuyuki, A., Lingna, Li., Kensei, K., Michael, B., Hoffman, R.M.GFP-expressing vascularization of Gelfoam® as a rapid in vivo assay of angiogenesis stimulators and inhibitors. Biotechniques. 2018; 42(3):294-298. DOI: https://doi.org/10.2144/000112382

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Published

2022-11-14

How to Cite

1.
Muhi Rasoul L, Ali Marhoon A, Fadhel Abbas Albaayit S, Waleed Ali R, Hadi Saleh T, Abdullah Laftah Al-Rubaii B. Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system. Biomedicine [Internet]. 2022 Nov. 14 [cited 2022 Nov. 27];42(5):938-42. Available from: https://biomedicineonline.org/home/article/view/1934

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