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在五种食管鳞状细胞癌细胞系中,p53的功能激活与其基因型无关。

p53 functional activation is independent of its genotype in five esophageal squamous cell carcinoma cell lines.

作者信息

Ji Junfang, Wu Kun, Wu Min, Zhan Qimin

机构信息

State Key Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100021, China.

出版信息

Front Med China. 2010 Dec;4(4):412-8. doi: 10.1007/s11684-010-0260-x. Epub 2010 Dec 30.

Abstract

p53 mutations have been found in many esophageal squamous cell carcinoma (ESCC) clinical specimens and cell lines. We reasoned that functional inactivation of wild-type p53 or the functional activation of mutant-type p53 might exist in these specimens and cell lines. In this study, we identified the correlation between p53 functional activation and its genotype in five different ESCC cell lines. To examine the potential p53 activation in a certain ESCC cell line, DNA damage methods including X-ray exposure and cisplatin treatment were employed to treat cells. Further, the expression of p53 protein and four transcripts of well-known p53 target genes were investigated using Western blot and reverse transcriptionpolymerase chain reaction (RT-PCR) after cell exposure to DNA damage. The results showed that in KYSE 30 cell line with mutant p53 and KYSE 150 with wild-type p53, p53 could be activated by DNA damages. However, p53 could not be activated following the DNA damages in YES 2 with wild-type p53, KYSE 70 with mutant p53, and EC9706 with unknown p53 genotype. All our data indicated that p53 function in certain cells is not closely correlated with its genotype. To judge p53 function in a particular cell line, it is important to examine the p53 functional activation, but not to simply rely on the p53 genotype.

摘要

在许多食管鳞状细胞癌(ESCC)临床标本和细胞系中都发现了p53突变。我们推断在这些标本和细胞系中可能存在野生型p53的功能失活或突变型p53的功能激活。在本研究中,我们确定了五种不同ESCC细胞系中p53功能激活与其基因型之间的相关性。为了检测某一ESCC细胞系中潜在的p53激活情况,采用包括X射线照射和顺铂处理在内的DNA损伤方法处理细胞。此外,在细胞暴露于DNA损伤后,使用蛋白质免疫印迹法和逆转录-聚合酶链反应(RT-PCR)研究p53蛋白的表达以及四个著名p53靶基因的转录本。结果显示,在具有突变型p53的KYSE 30细胞系和具有野生型p53的KYSE 150细胞系中,p53可被DNA损伤激活。然而,在具有野生型p53的YES 2、具有突变型p53的KYSE 70以及p53基因型未知的EC9706细胞系中,DNA损伤后p53未被激活。我们所有的数据表明,某些细胞中的p53功能与其基因型并非密切相关。要判断某一特定细胞系中的p53功能,检测p53的功能激活很重要,而不是仅仅依赖于p53基因型。

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