中国生物工程杂志

[ABSTRACT] Aim：to detect how Rb-deficiency will affect responses to TGF-β induced cell cycle arrest of hepatocyte. Methods: primary hepatocytes were isolated and cultured, and Rb-specific adenovirus siRNA was transformed into cells. Then TGF-β was added daily and cell growth and cell cycles variations. Western blot and FQ-RT-PCR were adopted to detect pRb, E2F, c-MYC and p16 expression changes. Results: Primary hepatocytes were isolated and infected withRb-specific siRNA adenovirus. In control cells, treatment with TGFβ prevented cells to enter S phase via decreased cMYC activity, activation of P16INK4A activity. In Rb-null hepatocytes, E2F and cMYC activity decreased slightly but P16INK4A was not activated and the great majority of cells continued cycling. Rb is therefore central to TGFβ-induced cell cycle arrest in hepatocytes. Conclusion: The present results show that otherwise genetically normal hepatocytes with disabled Rb genes respond less well to the antiproliferative effects of TGF-β.