p53 mutation profiling of multiple esophageal carcinoma using laser capture microdissection to demonstrate field carcinogenesis
p53 mutation profiling of multiple esophageal carcinoma using laser capture microdissection to demonstrate field carcinogenesis
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DOI：
10.1002/ijc.20500
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发表时间：
2005-01-01
影响因子：
6.4
通讯作者：
Maehara, Y
中科院分区：
文献类型：
--
作者：
Ito, S;Ohga, T;Maehara, Y
p53抑癌基因的失活是在人食管癌中观察到的最常见的遗传改变之一。在食管癌患者中，肿瘤的显著病理特征之一是在食管内存在多个病变。然而，涉及多发性病变发生的分子机制仍然难以捉摸。为了表征多种食管癌中p53的改变并研究其在癌发生中的作用，我们使用激光捕获显微切割对11例手术切除的人食管癌进行了p53免疫组织化学和p53突变分析：多发性食管癌9例，1例食管壁内转移灶，1例与主灶相邻的上皮内癌灶。在每例多发性食管癌患者中，我们检查了1个主要病变和1个代表性伴发病变的样本。用同样的方法对新鲜冷冻的正常组织和主要病变（整个肿瘤）的肿瘤组织样品进行分子分析。p53蛋白在11例患者的22个病灶中有16个（72.7%）有表达。正常食管组织中未发现p53突变。在9例多发性食管癌中，1例（11.1%）点突变发生在全癌，3例（33.3%）点突变发生在主癌和伴发癌中。显微切割肿瘤标本中，免疫组化结果与分子生物学分析结果的符合率为54.5%（12/22）。在8例未观察到p53突变的完整肿瘤中，2例在主要病变的显微切割肿瘤样品中发现p53突变。所有6例多发性食管癌在显微切割肿瘤样本中显示p53突变，其主要病变和伴随病变之间的p53突变状态不一致。与此相反，壁内转移病灶和上皮内癌相邻的主要病变显示p53突变模式相同的主要病变。总之，激光捕获显微切割DNA的分析是有用的表征异质性的p53基因突变的多个癌病变，不能准确地分析在整个食管肿瘤DNA。在食管癌多发病变中发现不同的p53基因突变，进一步证明了人食管癌的多中心或区域致癌作用。(C)2004 Wiley-Liss，Inc.
Inactivation of the p53 tumor suppressor gene is one of the most frequent genetic alterations observed in human esophageal carcinomas. In patients with esophageal carcinoma, one of the significant pathological features of the tumor is the presence of multiple lesions within the esophagus. However, the molecular mechanisms involved in the occurrence of multiple lesions have remained elusive. To characterize p53 alterations in multiple esophageal carcinomas and to study their roles in carcinogenesis, we performed p53 immunohistochemical and p53 mutation analyses using laser capture microdissection on surgically resected human esophageal carcinomas from 11 patients: 9 patients with multiple esophageal carcinomas, 1 with an intramural metastasis lesion within the esophagus and 1 with an intraepithelial carcinoma lesion contiguous to the main lesion. In each of the patients with multiple esophageal carcinomas, we examined samples from 1 main lesion and 1 representative concomitant lesion. Molecular analyses of samples from fresh-frozen normal tissues and tumor tissues of the main lesion (whole tumor) were also performed by the same method. p53 protein accumulation was observed in 16 (72.7%) of 22 lesions from the 11 cases. No p53 mutation was found in normal esophageal tissues. In the 9 cases of multiple esophageal carcinomas, point mutations were detected in the whole tumor in 1 (11.1%) case, in the microdissected tumor samples of main lesions in 3 (33.3%) cases and in the microdissected tumor samples of concomitant lesions in 3 (33.3%) cases. For the microdissected tumor samples, there was a 54.5% (12/22) concordance rate between the results of immunostaining and molecular analysis. In the 8 cases of whole tumors in which a p53 mutation was not observed, 2 cases revealed p53 mutation in the microdissected tumor samples of the main lesion. All 6 cases of multiple esophageal carcinomas that showed a p53 mutation in the microdissected tumor sample had a discordant p53 mutational status between the main and concomitant lesions. In contrast, both the intramural metastasis lesion and the intraepithelial carcinoma contiguous to the main lesion showed p53 mutational patterns identical to those of the main lesions. In conclusion, the analysis of microdissected DNA by laser capture microdissection is useful for characterizing the heterogeneity of the p53 gene mutation in multiple carcinoma lesions that cannot be accurately analyzed in whole esophageal tumor DNA. The finding of different p53 gene mutations among multiple esophageal carcinoma lesions suggest further evidence of multicentric or field carcinogenesis of the human esophagus. (C) 2004 Wiley-Liss, Inc.