Dynamics of subgenomic hepatitis C virus replicon RNA levels in Huh-7 cells after exposure to nucleoside antimetabolites.
Treatment with antimetabolites results in chemically induced low nucleoside triphosphate pools and cell cycle arrest in exponentially growing cells. Since steady-state levels of hepatitis C virus (HCV) replicon RNA were shown to be dependent on exponential growth of Huh-7 cells, the effects of antimetabolites for several nucleoside biosynthesis pathways on cell growth and HCV RNA levels were investigated. A specific anti-HCV replicon effect was defined as (i). minimal interference with the exponential cell growth, (ii). minimal reduction in cellular host RNA levels, and (iii). reduction of the HCV RNA copy number per cell compared to that of the untreated control. While most antimetabolites caused a cytostatic effect on cell growth, only inhibitors of the de novo pyrimidine ribonucleoside biosynthesis mimicked observations seen in confluent replicon cells, i.e., cytostasis combined with a sharp decrease in replicon copy number per cell. These results suggest that high levels of CTP and UTP are critical parameters for maintaining the steady-state level replication of HCV replicon in Huh-7 cells.
Stuyver
Lieven J
LJ
Pharmasset, Inc, Tucker, Georgia 30084, USA. lstuyver@pharmasset.com
McBrayer
Tamara R
TR
Tharnish
Phillip M
PM
Hassan
Abdalla E A
AE
Chu
Chung K
CK
Pankiewicz
Krzysztof W
KW
Watanabe
Kyochi A
KA
Schinazi
Raymond F
RF
Otto
Michael J
MJ
D000963:Antimetabolites / Q000494:pharmacology*; D000998:Antiviral Agents / Q000494:pharmacology*; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D009705:Nucleosides / Q000494:pharmacology*; D012367:RNA, Viral / Q000032:analysis*; D012093:Replicon / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D014779:Virus Replication / Q000187:drug effects
D000963:Antimetabolites; D000998:Antiviral Agents; D009705:Nucleosides; D012367:RNA, Viral