Chang Yu-Ching, Liao Chu-Bin, Hsieh Pei-Yu Chiang, Liou Ming-Li, Liu Yin-Chang
Institute of Molecular Medicine, National Tsing-Hua University, Hsin-Chu, Taiwan.
J Cell Biochem. 2008 Feb 1;103(2):528-37. doi: 10.1002/jcb.21428.
Tumor suppressor p53 is an essential regulator in mammalian cellular responses to DNA damage including cell cycle arrest and apoptosis. Our study with Chinese hamster ovary CHO-K1 cells indicates that when p53 expression and its transactivation capacity was inhibited by siRNA, UVC-induced G2/M arrest or apoptosis were unaffected as revealed by flow cyotmetric analyses and other measurements. However, inhibition of p53 rendered the cells slower to repair UV-induced damages upon a plasmid as shown in host cell reactivation assay. Furthermore, the nuclear extract (NE) of p53 siRNA-treated cells was inactive to excise the UV-induced DNA adducts as analyzed by comet assay. Consistently, the immunodepletion of p53 also deprived the excision activity of the NE in the similar experiment. Thus, tumor suppressor p53 of CHO-K1 cells may facilitate removal of UV-induced DNA damages partly via its involvement in the repair mechanism.
肿瘤抑制因子p53是哺乳动物细胞对DNA损伤(包括细胞周期停滞和凋亡)做出反应的重要调节因子。我们对中国仓鼠卵巢CHO-K1细胞的研究表明,当p53表达及其反式激活能力被小干扰RNA(siRNA)抑制时,如通过流式细胞术分析和其他测量所显示的,紫外线C(UVC)诱导的G2/M期停滞或凋亡不受影响。然而,如宿主细胞再激活试验所示,抑制p53使细胞在修复质粒上的紫外线诱导损伤时速度变慢。此外,通过彗星试验分析,用p53 siRNA处理的细胞的核提取物(NE)在切除紫外线诱导的DNA加合物方面没有活性。同样,在类似实验中,p53的免疫去除也剥夺了NE的切除活性。因此,CHO-K1细胞的肿瘤抑制因子p53可能部分通过参与修复机制促进紫外线诱导的DNA损伤的清除。