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增殖细胞核抗原(PCNA/细胞周期蛋白)免疫细胞化学作为小鼠肺组织中的一种标记指数

Proliferating cell nuclear antigen (PCNA/cyclin) immunocytochemistry as a labeling index in mouse lung tissues.

作者信息

Thaete L G, Ahnen D J, Malkinson A M

机构信息

Molecular and Environmental Toxicology Program, School of Pharmacy, University of Colorado, Boulder 80309-0297.

出版信息

Cell Tissue Res. 1989 Apr;256(1):167-73. doi: 10.1007/BF00224731.

Abstract

Proliferating cell nuclear antigen is expressed in cells from late G1 through the S-phase of the cell cycle. Therefore, antibodies directed against this molecule should provide a probe for labeling immunocytochemically the nuclei of proliferating cells. Herein we demonstrate the feasibility and reliability of this technique by quantifying immunostained pulmonary nuclei. We applied polyclonal and monoclonal antisera to alveolar and bronchiolar pulmonary epithelial cells in various proliferative states in tissue-sections and in vitro. A/J mice had a slightly higher labeling index than C57BL/6J mice, and proliferation in both strains increased dramatically after butylated hydroxytoluene treatment produced compensatory hyperplasia of Type-II pneumocytes. Immunostaining in fetal and neonatal lung samples from mice was higher than in adults. Spontaneous lung adenomas had a higher labeling index than the surrounding normal lung tissue. In addition, new data contained herein demonstrate a strain difference in proliferation of bronchiolar epithelial cells, and quantify the extent to which BHT-induced lung damage increases these proliferative rates. This mammalian nuclear antigen did not cross-react with antiserum to a functionally related bacterial protein, the beta subunit of E. coli DNA polymerase-III holoenzyme.

摘要

增殖细胞核抗原在细胞周期的G1晚期至S期的细胞中表达。因此,针对该分子的抗体应能提供一种用于免疫细胞化学标记增殖细胞核的探针。在此,我们通过对免疫染色的肺细胞核进行定量,证明了该技术的可行性和可靠性。我们将多克隆和单克隆抗血清应用于组织切片和体外处于各种增殖状态的肺泡和细支气管肺上皮细胞。A/J小鼠的标记指数略高于C57BL/6J小鼠,在丁基化羟基甲苯处理导致II型肺细胞代偿性增生后,两种品系的增殖均显著增加。小鼠胎儿和新生儿肺样本中的免疫染色高于成年小鼠。自发性肺腺瘤的标记指数高于周围正常肺组织。此外,本文包含的新数据表明细支气管上皮细胞增殖存在品系差异,并量化了丁基化羟基甲苯诱导的肺损伤增加这些增殖率的程度。这种哺乳动物核抗原与针对功能相关细菌蛋白(大肠杆菌DNA聚合酶III全酶的β亚基)的抗血清无交叉反应。

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