PAKISTAN
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Regulation of Pro-Apoptotic Bax and Trail Gene Expressions by H5N1 in Madin-Darby Canine Kidney (Mdck) Cell Line
 
Wei Boon Yap1,2*, Kah Chun Goh1, Sharifah Syed Hassan3 and Vinod RMT Balasubramaniam3
 
1Biomedical Science Programme, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia; 2Center of Toxicology and Health Risk Studies, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia; 3Jeffrey Cheah School of Medicine & Health Sciences, Monash University Malaysia, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia
*Corresponding author: yapweiboon@ukm.edu.my

Abstract   

Avian influenza virus (AIV) A H5N1 is able to cause zoonosis with high morbidity and mortality. Several studies suggested the regulation of cellular apoptosis for virus survival by influenza viruses. This sheds light on the development of antivirals that can improve cellular apoptosis to fight influenza infections. This study aimed to investigate the regulation of pro-apoptotic gene expressions by H5N1 in MDCK cells. The H5N1 infection (102.67TCID50/ml) was performed for 8, 24, 48, 72, 96 and 120 hours. Upon incubation, the percentage of cell death, DNA fragmentation and pro-apoptotic gene expressions i.e. Bax, TRAIL, Caspase 3 (Cas3) and 8 (Cas8), and Fas-ligand (FasL) were determined. The degree of DNA laddering and  pro-apoptotic gene expressions were determined on agarose gels. The results indicated that severe cytopathic effects (CPE) caused by H5N1 appeared as early as 24 hour post-infection (hpi) meanwhile a significant cell death was observed starting at 48 hpi. However, DNA laddering was not observed in both of the control and infected DNA samples. Downregulation of Bax gene expression was recorded at 24 -72 hpi and a total suppression was observed at 96 and 120 hpi. Similarly, a significant decrease in the TRAIL gene expression was observed at 48-120 hpi. On the contrary, the expressions of Cas3, Cas8 and FasL genes were not detected in the mock and infected samples. Conclusively, this study suggested that H5N1 might downregulate cellular apoptosis prematurely for viral survival which can be a target cellular mechanism for newly developed antivirals to combat H5N1 infections.

To Cite This Article: Yap WB, Goh KC, Hassan SS, Balasubramaniam VRMT 2020. Regulation of pro-apoptotic bax and trail gene expressions by H5N1 in madin-darby canine kidney (Mdck) cell line. Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2020.055

 
   

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)



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