Human Cytomegalovirus Replication in Human Embryonic Fibroblast Transformed by 60 Coγ Irradiation

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Journal Title, Volume, Page: 
Palestinian Med. J.; 1 (2): 72-77
Year of Publication: 
Bashar Saad
Current Affiliation: 
Faculty of Medicine & Health Sciences, Department of Biomedical Sciences, An-Najah National University, Nablus, Palestine
Preferred Abstract (Original): 

The replication of human cytomegalovirus (HCMV) was studied in human embryonic cells transformed and immortalized by 60Co g irradiation (KMST-6).The HCMV production in KMST-6 cells was delayed when compared to the virus production in normal human embryonic (KMS-6) cells.Growth studies revealed that virus titer at 5 days postinfection (pi) in KMST-6 was more than 5 logs less than KMS-6 cells.Western blot analysis showed that the reduction of the viral titer was due to delay in the major immediate early (MIE), mainly MIE2, and consequently the early and late protein synthesis.  On the cellular level, HCMV mediated c-Jun, c-Fos and NFkB activities, which are necessary for MIE protein synthesis, were induced in KMS-6 but not in KMST-6 cells.In the contrary with KMS-6 cells, treating KMST-6 cells with LY294002- an inhibitor of cellular phosphatidylinositol 3-kinase (PI3-K) - enhanced virus protein synthesis and virus replication by 3 logs at 5 days pi. RT-PCR and electrophoretic mobility shift assay indicated that LY294002 activated the MIE protein synthesis through the activation of NFkB and MIE gene expression.These results suggest that transforming the embryonic fibroblast may cause the disruption of the down stream of PI3-K signaling pathway leading to the activation of c-Jun, c-Fos and NFkB, which play a crucial role for expression of the critical MIE genes.