IMMUNOHISTOCHEMICAL ANALYSIS OF THE P53 ONCOPROTEIN ON PARAFFIN SECTIONS USING A SERIES OF NOVEL MONOCLONAL-ANTIBODIES
IMMUNOHISTOCHEMICAL ANALYSIS OF THE P53 ONCOPROTEIN ON PARAFFIN SECTIONS USING A SERIES OF NOVEL MONOCLONAL-ANTIBODIES
复制标题
DOI：
10.1002/path.1711690106
复制
发表时间：
1993-01-01
影响因子：
7.3
通讯作者：
LANE, DP
中科院分区：
文献类型：
--
作者：
BARTEK, J;BARTKOVA, J;LANE, DP
p53肿瘤抑制基因的改变被认为是多种人类癌症的多阶段癌变过程中的关键事件。为了阐明各种p53突变在肿瘤发生中的作用，并研究它们与p53蛋白积累和亚细胞定位的关系，我们提出了一系列新的21种小鼠抗人重组p53单克隆抗体（MAbs）。新的单克隆抗体（命名为Bp 53系列）似乎主要识别抗变性抗原表位的免疫印迹和他们中的大多数是合适的p53在培养细胞和冷冻切片的免疫染色。此外，至少有三种单克隆抗体（Bp 53 -11，Bp 53 -12，和Bp 53 -28）被证明是可靠的试剂，用于石蜡包埋标本的免疫组化。用Bp 53 -11和Bp 53 -12对118例不同组织发生的人类肿瘤的石蜡切片进行免疫组织化学分析，结果显示76例（64%）肿瘤细胞中不同比例的p53蛋白在核内积聚。组织处理的三个参数（固定剂的类型，固定时间，自溶持续时间）对p53蛋白检测的影响也进行了研究。本研究的结果提供了必要的基础，更广泛地应用这些新的单克隆抗体作为工具，在常规的组织病理学和功能分析的p53癌蛋白。
Alterations of the p53 tumour suppressor gene are considered critical events in multistage carcinogenesis of a wide range of human cancers. In an attempt to elucidate the role of various p53 mutations in tumorigenesis and to investigate their relationship to the p53 protein accumulation and subcellular localization, we have raised a new series of 21 mouse monoclonal antibodies (MAbs) to human recombinant p53. The new MAbs (designated the Bp53 series) appear to recognize mainly denaturation-resistant epitopes in immunoblotting and the majority of them are suitable for immunostaining of p53 in cultured cells and frozen sections. Furthermore, at least three MAbs (Bp53-11, Bp53-12, and Bp53-28)proved to be reliable reagents for immunohistochemistry on paraffin-embedded specimens. The immunohistochemical analysis of paraffin sections from 118 human tumours of various histogeneses with Bp53-11 and Bp53-12 showed nuclear accumulation of the p53 protein in variable proportion of tumour cells in 76 cases (64 per cent). The influence of three parameters of tissue processing (type of fixative, period of fixation, and duration of autolysis) on p53 protein detection was also investigated. The results of this study provide the necessary basis for wider application of these novel MAbs as tools in both routine histopathology and functional analyses of the p53 oncoprotein.