p53 gene and cell cycling in uveal melanoma
p53 gene and cell cycling in uveal melanoma
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DOI：
10.1016/s0002-9394(14)75431-5
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发表时间：
1996-05-01
影响因子：
4.2
通讯作者：
Selle, J
中科院分区：
文献类型：
--
作者：
Kishore, K;Ghazvini, S;Selle, J
目的：确定葡萄膜黑色素瘤中是否存在肿瘤抑制基因 p53 的改变，并进一步表征这些变化的性质。方法：使用单克隆抗体进行免疫组织化学分析，以确定 35 个去核档案葡萄膜黑色素瘤中是否存在 p53 的改变。通过使用溴脱氧尿苷染色比较 p53 基因和细胞周期状态来进行进一步表征，使用聚合酶链反应单链构象多态性分析和测序来表征 p53 的改变。 结果：使用单克隆抗体 DO-7，35 个葡萄膜黑色素瘤中的 4 个肿瘤细胞核中 p53 改变的阳性免疫染色呈低水平（0.5％至 5.0％），这四个肿瘤具有三个最高水平溴脱氧尿苷标记指数排名第 14，范围为 1.3% 至 7.0%。对三个 p53 阳性和六个 p53 阴性肿瘤进行 p53 外显子 5 至 8 的聚合酶链反应单链构象多态性分析，未检测到运动变化的改变。对其中一份阳性染色样本进行测序证实 p53 基因的外显子 5 至 8 没有突变。对一个肿瘤中的溴脱氧尿苷和p53进行双重免疫组织化学标记，结果显示大多数p53阳性细胞处于S期。结论：p53突变在葡萄膜黑色素瘤中是一种罕见的事件。在细胞周期水平最高的四个肿瘤中的三个中发现了 p53 蛋白的核积累。
PURPOSE: To determine whether alterations of p53, a tumor suppressor gene, were present in uveal melanoma, and to characterize further the nature of those changes.METHODS: Immunohistochemical analysis with a monoclonal antibody was used to determine whether alterations of p53 were present in 35 enucleated archival uveal melanomas. Further characterization was done by comparing the p53 gene and cell cycling status by using bromodeoxyuridine staining, The alterations in p53 were characterized using polymerase chain reaction single-strand conformational polymorphism analysis and sequencing.RESULTS: Four of 35 uveal melanomas showed low levels (0.5% to 5.0%) of positive immunostaining for altered p53 in tumor cell nuclei using monoclonal antibody DO-7, These four tumors had the three highest and the 14th highest bromodeoxyuridine labeling indices, ranging from 1.3% to 7.0%. Polymerase chain reaction single-strand conformational polymorphism analysis of p53 exons 5 to 8 was performed on three p53-positive and six p53-negative tumors, and no altered motility shifts were detected. Sequencing of one of the positive staining specimens confirmed no mutations in exons 5 through 8 in the p53 gene. Double immunohistochemical labeling for both bromodeoxyuridine and p53 in one tumor showed that most of p53-positive cells were in S phase.CONCLUSIONS: Mutation of p53 is an uncommon event in uveal melanomas. Nuclear accumulation of p53 protein was found in three of the four tumors with the highest levels of cell cycling.