Institute of Environmental Toxicology, Martin Luther University Halle Wittenberg, Franzosenweg 1a, 06097, Halle, Saale, Germany.
Arch Toxicol. 2011 Jun;85(6):669-79. doi: 10.1007/s00204-010-0604-y. Epub 2010 Oct 16.
Activation of poly(ADP-ribose) polymerase-1 (PARP-1) in response to DNA damage is an important mechanism to keep homeostasis or to trigger apoptosis. The expression and function of (PARP-1) was studied in primary cells cultured from human lung. Normal human bronchial epithelial cells (NHBEC) and peripheral lung cells (PLC) from lung cancer patients were grown as explant cultures and were followed over a period of 12 weeks. PARP-1 protein was expressed in all explant cultures from bronchial epithelium. The levels of PARP protein differed between individuals by a factor of 2.3 in the first explant. Three cases were followed for more than 100 days. The expression levels varied intra-individually by a factor of 1.3-1.4 over this time period. PARP-1 activity was determined immunohistochemically after induction of DNA damage with H(2)O(2) (0.05-0.3 mM, 5 min). The fluorescence signal for ADP-ribose polymers attached to chromatin proteins correlated well with the concentration of H(2)O(2). PARP-1 activity differed by a factor of 3.1 in NHBECs obtained from the first generation of explants from 11 cases. PARP-1 activity is present in NHBECs until the 8th and in PLCs until the 12th week and declined to about half of the start level. Primary cultures of NHBECs and PLC are suitable to study the effect of external factors on PARP-1 expression and function.
聚 ADP-核糖聚合酶-1（PARP-1）在 DNA 损伤时的激活是维持内稳态或触发细胞凋亡的重要机制。我们研究了来源于人肺的原代细胞中 PARP-1 的表达和功能。正常的人支气管上皮细胞（NHBEC）和肺癌患者的肺外周细胞（PLC）作为外植体培养，并在 12 周内进行了跟踪。PARP-1 蛋白在所有支气管上皮的外植体培养物中都有表达。在最初的外植体中，PARP 蛋白的个体间水平差异高达 2.3 倍。有 3 例患者的随访时间超过 100 天。在此期间，个体内的表达水平差异为 1.3-1.4 倍。用 H2O2（0.05-0.3mM，5 分钟）诱导 DNA 损伤后，通过免疫组化测定 PARP-1 活性。与 H2O2 浓度相关的附着于染色质蛋白的 ADP-核糖聚合物的荧光信号。11 例第一代外植体的 NHBEC 中 PARP-1 活性的差异高达 3.1 倍。PARP-1 活性在 NHBEC 中持续存在到第 8 周，在 PLC 中持续存在到第 12 周，然后下降到起始水平的一半左右。NHBEC 和 PLC 的原代培养物适合研究外部因素对 PARP-1 表达和功能的影响。
