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四种人髓母细胞瘤衍生细胞系中的p53表达

p53 expression in four human medulloblastoma-derived cell lines.

作者信息

Srinivasan J, Berger M S, Silber J R

机构信息

Department of Neurological Surgery, University of Washington Medical Center, Seattle 98195, USA.

出版信息

Childs Nerv Syst. 1996 Feb;12(2):76-80. doi: 10.1007/BF00819500.

Abstract

p53 is a tumor suppressor gene found on the short arm of chromosome 17. Loss of one p53 allele and alteration of the other is found in a variety of tumors, including highgrade glial tumors. Point mutations in the remaining allele occur in a highly conserved region of the gene encompassing exons 5-8. Although 40-50% of medulloblastomas lose sequences on the short arm of chromosome 17, alteration of p53 in these tumors is infrequent. To further characterize genetic alteration of p53 in medulloblastoma, we performed a mutational analysis of four medulloblastoma-derived cell lines established by our laboratory. Using two variable-number tandem repeat markers which map distally to p53, we found evidence indicating loss of sequences on the distal end of chromosome 17p in all four lines. However, no gross alterations of the p53 gene were detected. Northern analysis revealed expression of equivalent amounts of full-length p53 messenger RNA in each cell line. Using the polymerase chain reaction to amplify exons 5-8 of the p53 gene, we directly sequenced the amplified fragments and detected no mutations in any of the cell lines. Our results demonstrate that loss of p53 function through gene deletion and/or recessive mutation is not required for growth in our cell lines.

摘要

p53是一种肿瘤抑制基因,位于17号染色体短臂上。在包括高级别胶质瘤在内的多种肿瘤中都发现一个p53等位基因缺失,另一个发生改变。剩余等位基因的点突变发生在该基因包含外显子5 - 8的高度保守区域。虽然40% - 50%的髓母细胞瘤在17号染色体短臂上存在序列缺失,但这些肿瘤中p53的改变并不常见。为了进一步明确髓母细胞瘤中p53的基因改变情况,我们对本实验室建立的4个髓母细胞瘤衍生细胞系进行了突变分析。使用两个定位于p53远端的可变数目串联重复标记,我们发现所有4个细胞系中均有17号染色体短臂远端序列缺失的证据。然而,未检测到p53基因的明显改变。Northern印迹分析显示每个细胞系中全长p53信使核糖核酸的表达量相当。使用聚合酶链反应扩增p53基因的外显子5 - 8,我们对扩增片段进行直接测序,未在任何细胞系中检测到突变。我们的结果表明,在我们的细胞系中,通过基因缺失和/或隐性突变导致p53功能丧失并非细胞生长所必需。

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