Expression and sequence analysis of the p21(WAF1/CIP1) gene in renal cancers
Expression and sequence analysis of the p21(WAF1/CIP1) gene in renal cancers
复制标题
DOI：
10.1016/s0090-4295(96)00493-1
复制
发表时间：
1997-03-01
期刊：
影响因子：
2.1
通讯作者：
Nanus, DM
中科院分区：
文献类型：
--
作者：
Papandreou, CN;Bogenrieder, T;Nanus, DM
目标。p21 （WAF1/CIP1）周期蛋白依赖性激酶抑制剂是一种M(r) 21,000蛋白，可以通过与细胞退出G所需的周期蛋白依赖性激酶复合物相关联并抑制其抑制细胞生长(1)。它是p53生长控制通路的下游效应物，可以通过增加p53蛋白水平来转录激活。本研究的目的是确定肾癌中p21表达的突变或改变是否有助于肾癌细胞的生长。采用单链构象多态性分析和直接脱氧核糖核酸（DNA）测序技术检测了12株肾癌细胞系p21基因编码区的突变。12株细胞系中p21的表达分别采用cDNA探针和p21特异性抗体检测。在两个细胞系中检测到p21基因的核苷酸碱基替换，但不导致氨基酸替换。Northern分析发现，12种肾癌细胞系中有8种存在p21特异性mRNA，尽管在所有12种肾癌中均可通过聚合酶链反应检测到p21转录本。12例肾癌中有9例检测到不同水平的p21特异性蛋白。这些数据表明，p21基因的突变在肾癌细胞系中是罕见的，肾癌细胞的不受控制的生长不是由于p21基因的突变。然而，表达研究发现p21蛋白水平在肾癌细胞中有广泛的变化，提示p21表达的异常调节可能在肾癌的发展中起作用。(C) 1997, Elsevier Science Inc.；
Objectives. The p21 (WAF1/CIP1) cyclin-dependent kinase inhibitor is an M(r) 21,000 protein that can arrest cell growth by associating with and inhibiting cyclin-dependent kinase complexes necessary for cells to exit G(1). It is a downstream effector in the p53 growth control pathway and can be transcriptionally activated by increasing levels of p53 protein. The objective of this study was to determine if there are mutations or alterations in the expression of p21 in renal cancers that could contribute to renal cancer cell growth.Methods. Twelve renal cancer cell lines were examined for mutations in the coding region of the p21 gene using single-stranded conformation polymorphism analysis and direct deoxyribonucleic acid (DNA) sequencing. Expression of p21 was determined in all 12 cell lines by Northern analysis using a cDNA probe for p21 and Western analyses using a p21-specific antibody.Results. Nucleotide base substitutions were detected in the p21 gene in two cell lines, which did not result in amino acid substitutions. P21-specific mRNA was present in 8 of 12 renal cancer cell lines, as determined by Northern analysis, although p21 transcripts could be detected by polymerase chain reaction in all 12 renal cancers. Varying levels of p21-specific protein were detected in 9 of 12 renal cancers.Conclusions. These data indicate that mutation of the p21 gene is rare in renal cancer cell lines and that the uncontrolled growth of renal cancer cells is not due to mutation of the p21 gene. However, expression studies found a wide variation in the level of p21 protein in renal cancer cells, suggesting that aberrant regulation of p21 expression may play a role in renal cancer development. (C) 1997, Elsevier Science Inc.