Small molecular weight variants of p53 are expressed in human melanoma cells and are induced by the DNA-damaging agent cisplatin
Small molecular weight variants of p53 are expressed in human melanoma cells and are induced by the DNA-damaging agent cisplatin
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DOI：
10.1158/1078-0432.ccr-07-1422
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发表时间：
2008-03-15
影响因子：
11.5
通讯作者：
Hersey, Peter
中科院分区：
文献类型：
--
作者：
Avery-Kiejda, Kelly A.;Zhang, Xu Dong;Hersey, Peter
转移性黑色素瘤对DNA损伤性化疗药物基本上无反应，尽管WTp 53经常被检测到。已经发现了几种p53的同种型，其中一些抑制p53功能。因此，我们研究了是否p53亚型存在于黑色素瘤，以及他们是否可能有助于异常p53功能在melanoma.Experimental Designing：我们研究了p53及其亚型的表达和亚细胞定位在一个小组的人黑色素瘤细胞系使用Western印迹，双向电泳，逆转录-PCR。我们还描述了DNA损伤剂顺铂治疗后p53、p53亚型和p53靶基因表达之间的关系。我们报道了p53和Delta 40 p53在大多数黑素瘤细胞系中以mRNA水平表达，但在成纤维细胞和黑素细胞中不存在或以低水平表达，提示它们的表达可能在黑色素瘤的发展中起作用。双向凝胶电泳分析显示，p53 β在黑色素瘤细胞中的蛋白质水平上表达。与正常成纤维细胞相比，p53和小分子量形式的p53在黑色素瘤细胞系的核和胞质组分之间异常表达。用顺铂治疗对WTP 53和小分子量形式的p53具有细胞系依赖性的不同影响。三角洲40 p53被证明抑制，而p53被证明增强，p53依赖的转录p21和p53。结论：p53和三角洲40 p53在黑色素瘤中表达，这可能有重要的意义，了解黑色素瘤的耐药性DNA损伤化疗。
Purpose: Metastatic melanoma is largely unresponsive to DNA-damaging chemotherapy agents, although WTp53 is frequently detected. Several isoforms of p53 have been discovered, some of which inhibit p53 function. We therefore examined whether p53 isoforms were present in melanoma and whether they may contribute to aberrant p53 function in melanoma.Experimental Design: We studied the expression and subcellular localization of p53 and its isoforms in a panel of human melanoma cell lines using Western blot, two-dimensional electrophoresis, and reverse transcription-PCR. We also characterized the relationship between the expression of p53, p53 isoforms, and p53 target genes following treatment with the DNA-damaging agent cisplatin.Results: We report that p53 and Delta 40p53 were expressed in the majority of melanoma cell lines at the mRNA level, but were absent or expressed at low levels in fibroblasts and melanocytes, suggesting that their expression may play a role in melanoma development. Analysis by two-dimensional gel electrophoresis revealed that p53 beta was expressed at the protein level in melanoma cells. Both p53 and the small molecular weight forms of p53 were aberrantly expressed between the nuclear and cytosolic fractions of melanoma cell lines, compared with normal fibroblasts. Treatment with cisplatin had differential effects on WTp53 and the small molecular weight form of p53 that were cell line dependent. Delta 40p53 was shown to inhibit, whereas p53 was shown to enhance, p53-dependent transcription of p21 and PUMA.Conclusions: p53 and Delta 40p53 are expressed in melanoma and this may have important implications for understanding resistance of melanoma to DNA-damaging chemotherapy.