Harashima Mizuho, Seki Taiichiro, Ariga Toyohiko, Niimi Shingo
Department of Nutrition and Physiology, Nihon University College of Bioresource Sciences, Kameino Fujisawa 252-8510, Japan.
Biomed Res. 2013;34(5):269-73. doi: 10.2220/biomedres.34.269.
In the present study, we investigated the role of p16(INK4a) in the inhibition of DNA synthesis stimulated by hepatocyte growth factor (HGF) or epidermal growth factor (EGF) using RNA interference in primary cultured rat hepatocytes. The transfection of small interfering RNAs targeting p16(INK4a) reduced the corresponding mRNA and protein expression by more than approximately 90% and 50%, respectively, at 24 h after transfection. In the cells transfected with p16(INK4a) small interfering RNA, control, HGF, and EGF-stimulated DNA synthesis as assessed by (3)H-thymidine incorporation increased by approximately 1.5-fold, 1.6-fold, and 1.7-fold, respectively, compared with that in the control small interfering RNA-transfected cells. These findings indicate that p16(INK4a) plays a significant role in the inhibition of DNA synthesis.
在本研究中，我们利用RNA干扰技术，在原代培养的大鼠肝细胞中研究了p16(INK4a)在抑制由肝细胞生长因子(HGF)或表皮生长因子(EGF)刺激的DNA合成中的作用。靶向p16(INK4a)的小干扰RNA转染后24小时，相应的mRNA和蛋白表达分别降低了约90%和50%以上。在用p16(INK4a)小干扰RNA转染的细胞中，通过³H-胸苷掺入评估的对照、HGF和EGF刺激的DNA合成，与对照小干扰RNA转染的细胞相比，分别增加了约1.5倍、1.6倍和1.7倍。这些发现表明，p16(INK4a)在抑制DNA合成中起重要作用。
