Blunted DNA synthesis and delayed S-phase entry following inhibition of Cdk2 activity in the regenerating rat liver

Activation of the cyclin E/Cdk2 complex may play an important role in mid-G1/S-phase progression in proliferating mammalian cells. We evaluated the effect of targeted inhibition of Cdk2 activity by CYC202 (R-roscovitine) on hepatocytes proliferation in vivo after 70% partial hepatectomy (PH) in rats. In controls, Cdk2 activity and DNA synthesis peaked 24 h after PH. CYC202 abrogated Cdk2 activity, prevented BrdU incorporation and PCNA expression and increased mortality 24 h after PH. Cyclin E and Cdk2 protein expression and complex formation was not affected by CYC202 nor was cyclin D1, Cdk4 and c-ras mRNA expression. Two consecutive injections 8 and 20 h after PH were required to elicit the inhibitory effect of CYC202, which was lost when either the injection at 8 h or at 20 h was withheld. Cdk2 activity and cell progression resumed 48 h after PH in surviving animals suggesting that CYC202 induced a reversible inhibition of the cell cycle. Our results confirm an important role for Cdk2 in hepatocytes proliferation in the regenerating liver. We demonstrate that molecular events, including Cdk2 activation, occurring within the 8th and 24th hour after PH (G1/S-phase transition) are crucial in determining whether or not DNA synthesis and hepatocytes proliferation proceed normally after PH.