Nutlin-3a activates p53 to both down-regulate inhibitor of growth 2 and up-regulate mir-34a, mir-34b, and mir-34c expression, and induce senescence

被引:189
作者
Kumamoto, Kensuke [1 ,4 ]
Spillare, Elisa A. [1 ]
Fujita, Kaori [1 ]
Horikawa, Izumi [1 ]
Yamashita, Taro [1 ]
Appella, Ettore [2 ]
Nagashima, Makoto [3 ]
Takenoshita, Seiichi [4 ]
Yokota, Jun [5 ]
Harris, Curtis C. [1 ]
机构
[1] NCI, Human Carcinogenesis Lab, Ctr Canc Res, NIH, Bethesda, MD 20892 USA
[2] NCI, Cell Biol Lab, Ctr Canc Res, NIH, Bethesda, MD 20892 USA
[3] Toho Univ, Sakura Hosp, Dept Surg, Sakura, Japan
[4] Fukushima Med Univ, Sch Med, Dept Surg 2, Fukushima, Japan
[5] Natl Canc Ctr, Res Inst, Div Biol, Tokyo 104, Japan
关键词
D O I
10.1158/0008-5472.CAN-07-2780
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Nutlin-3, an MDM2 inhibitor, activates p53, resulting in several types of cancer cells undergoing apoptosis. Although p53 is mutated or deleted in similar to 50%o of all cancers, p53 is still functionally active in the other 50%. Consequently, nutlin-3 and similar drugs could be candidates for neoadjuvant therapy in cancers with a functional p53. Cellular senescence is also a phenotype induced by p53 activation and plays a critical role in protecting against tumor development. In this report, we found that nutlin-3a can induce senescence in normal human fibroblasts. Nutlin-3a activated and repressed a large number of p53-dependent genes, including those encoding microRNAs. mir-34a, mir-34b, and mir-34c, which have recently been shown to be downstream effectors of p53-mediated senescence, were up-regulated, and inhibitor of growth 2 (ING2) expression was suppressed by nutlin-3a treatment. Two candidates for a p53-DNA binding consensus sequence were found in the ING2 promoter regulatory region; thus, we performed chromatin immunoprecipitation and electrophoretic mobility shift assays and confirmed p53 binding directly to those sites. In addition, the luciferase activity of a construct containing the ING2 regulatory region was repressed after p53 activation. Antisense knockdown of ING2 induces p53-independent senescence, whereas overexpression of ING2 induces p53-dependent senescence. Taken together, we conclude that nutlin-3a induces senescence through p53 activation in normal human fibroblasts, and p53-mediated mir34a, mir34b, and mir34c up-regulation and ING2 down-regulation may be involved in the senescence pathway.
引用
收藏
页码:3193 / 3203
页数:11
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