Apoptosis in dengue virus infected liver cell lines HepG2 and Hep3B.
While both in vivo and in vitro evidence has suggested that liver cells undergo apoptosis in response to dengue virus infection, little is known about the mechanism of induction. Given that the p53 tumour suppressor gene is a key mediator of apoptosis, we sought to define the role of this gene in response to dengue virus infection. After infection, a p53 wild type liver cell line (HepG2) showed changes consistent with apoptosis including alterations of cell morphology, cellular detachment and DNA laddering. However, p53 was neither up-regulated, nor showed any evidence of complexing with dengue virus proteins as determined by immunoprecipitation. Infection of a p53 null liver cell line (Hep3B) also produced changes consistent with the induction of apoptosis. While the profile of the cells undergoing apoptosis in each cell line was similar as determined by flow cytometry, the absolute levels were markedly different with up to 90% of Hep3B cells undergoing apoptosis compared to only 20% of HepG2 cells at day 5 post infection. By day 7, all Hep3B infected cells were dead. In contrast, it proved possible to culture dengue virus infected HepG2 cells for 3 months. Viral progeny released from the p53 null cell line were nine-fold higher per attached cell than from the p53 wild type cell line. These results suggest that, while induction of apoptosis in liver cells is mediated by a non-p53 regulated pathway, p53 may play a role in restricting the level of viral progeny to below a critical level at which apoptosis is triggered.
Thongtan
Thananya
T
Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, Nakhon Pathom, Thailand.
Panyim
Sakol
S
Smith
Duncan R
DR
D017209:Apoptosis; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D003716:Dengue Virus / Q000254:growth & development / Q000472:pathogenicity*; D017353:Gene Deletion; D005786:Gene Expression Regulation; D016158:Genes, p53; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D006801:Humans; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000378:metabolism
D016159:Tumor Suppressor Protein p53