PARP-1 expression and activity in primary human lung cells
PARP-1 expression and activity in primary human lung cells
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DOI：
10.1007/s00204-010-0604-y
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发表时间：
2011-06-01
影响因子：
6.1
通讯作者：
Foth, Heidi
中科院分区：
文献类型：
--
作者：
Ahmad, Mohamed;Torky, Abdelrahman;Foth, Heidi
聚腺苷二磷酸核糖聚合酶-1（PARP-1）在DNA损伤后的激活是维持体内稳态或触发细胞凋亡的重要机制。本研究在人肺原代培养细胞中研究了PARP-1的表达和功能。正常人支气管上皮细胞（NHBEC）和肺癌患者的外周肺细胞（PLC）作为外植体培养物生长，并随访12周。PARP-1蛋白在支气管上皮的所有外植体培养物中表达。PARP蛋白的水平在第一外植体中个体之间差异为2.3倍。3例随访时间超过100天。在这段时间内，表达水平个体内变化1.3-1.4倍。在用H2 O2（0.05-0.3 mM，5 min）诱导DNA损伤后，用化学方法测定PARP-1活性。ADP-核糖聚合物连接到染色质蛋白的荧光信号与H2 O2的浓度相关。从11例第一代外植体获得的NHBEC中PARP-1活性相差3.1倍。PARP-1活性存在于NHBEC中直到第8周，并且存在于PLC中直到第12周，并且下降至约起始水平的一半。NHBECs和PLC的原代培养适合于研究外部因素对PARP-1表达和功能的影响。
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 H2O2 (0.05-0.3 mM, 5 min). The fluorescence signal for ADP-ribose polymers attached to chromatin proteins correlated well with the concentration of H2O2. 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.