p63-microRNA feedback in keratinocyte senescence
p63-microRNA feedback in keratinocyte senescence
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DOI：
10.1073/pnas.1112257109
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发表时间：
2012-01-24
影响因子：
11.1
通讯作者：
Melino, Gerry
中科院分区：
文献类型：
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作者：
Cervo, Pia Rivetti di Val;Lena, Anna Maria;Melino, Gerry
我们研究了人原代角质形成细胞中与复制性衰老相关的microRNA（miRNAs）的表达。通过全基因组miRNA谱鉴定了衰老中上调的一组miRNA，并在增殖细胞与衰老细胞中验证了其表达变化。其中，miRNA（miR）-138、-181 a、-181 b和-130 b的表达随着连续传代而增加。增殖细胞中的miR-138、-181 a和-181 b过表达本身足以诱导衰老，这通过抑制BrdU掺入和定量衰老激活的β-半乳糖苷酶染色来评估。我们确定Sirt 1是miR-138、-181a和-181b的直接靶点，而Delta Np 63的表达被miR-130 b抑制。我们还发现，Delta Np 63 alpha通过直接结合位于原代角质形成细胞中这些miRNA基因组位点附近的p63响应元件来抑制miR-138、-181a、-181b和-130b表达。这些发现表明，通过调节调节蛋白如p63和Sirt 1的水平，miRNA表达的变化强烈地促进了原代人角质形成细胞衰老的诱导，从而将这两种蛋白质连接起来。我们的数据还表明，抑制miR-138、-181a、-181b和-130b表达是Delta Np 63 α在表皮增殖细胞中促进生长策略的一部分。
We investigated the expression of microRNAs (miRNAs) associated with replicative senescence in human primary keratinocytes. A cohort of miRNAs up-regulated in senescence was identified by genome-wide miRNA profiling, and their change in expression was validated in proliferative versus senescent cells. Among these, miRNA (miR)-138, -181a, -181b, and -130b expression increased with serial passages. miR-138, -181a, and -181b, but not miR-130b, overexpression in proliferating cells was sufficient per se to induce senescence, as evaluated by inhibition of BrdU incorporation and quantification of senescence-activated beta-galactosidase staining. We identified Sirt1 as a direct target of miR-138, -181a, and -181b, whereas Delta Np63 expression was inhibited by miR-130b. We also found that Delta Np63 alpha inhibits miR-138, -181a, -181b, and -130b expression by binding directly to p63-responsive elements located in close proximity to the genomic loci of these miRNAs in primary keratinocytes. These findings suggest that changes in miRNA expression, by modulating the levels of regulatory proteins such as p63 and Sirt1, strongly contribute to induction of senescence in primary human keratinocytes, thus linking these two proteins. Our data also indicate that suppression of miR-138, -181a, -181b, and -130b expression is part of a growth-promoting strategy of Delta Np63 alpha in epidermal proliferating cells.