Multifactorial ERβ and NOTCH1 control of squamous differentiation and cancer
Yang Sui Brooks, … , Karine Lefort, G. Paolo Dotto
Yang Sui Brooks, … , Karine Lefort, G. Paolo Dotto
Published April 17, 2014
Citation Information: J Clin Invest. 2014;124(5):2260-2276. https://doi.org/10.1172/JCI72718.
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Research Article Oncology

Multifactorial ERβ and NOTCH1 control of squamous differentiation and cancer

Abstract

Downmodulation or loss-of-function mutations of the gene encoding NOTCH1 are associated with dysfunctional squamous cell differentiation and development of squamous cell carcinoma (SCC) in skin and internal organs. While NOTCH1 receptor activation has been well characterized, little is known about how NOTCH1 gene transcription is regulated. Using bioinformatics and functional screening approaches, we identified several regulators of the NOTCH1 gene in keratinocytes, with the transcription factors DLX5 and EGR3 and estrogen receptor β (ERβ) directly controlling its expression in differentiation. DLX5 and ERG3 are required for RNA polymerase II (PolII) recruitment to the NOTCH1 locus, while ERβ controls NOTCH1 transcription through RNA PolII pause release. Expression of several identified NOTCH1 regulators, including ERβ, is frequently compromised in skin, head and neck, and lung SCCs and SCC-derived cell lines. Furthermore, a keratinocyte ERβ–dependent program of gene expression is subverted in SCCs from various body sites, and there are consistent differences in mutation and gene-expression signatures of head and neck and lung SCCs in female versus male patients. Experimentally increased ERβ expression or treatment with ERβ agonists inhibited proliferation of SCC cells and promoted NOTCH1 expression and squamous differentiation both in vitro and in mouse xenotransplants. Our data identify a link between transcriptional control of NOTCH1 expression and the estrogen response in keratinocytes, with implications for differentiation therapy of squamous cancer.

Authors

Yang Sui Brooks, Paola Ostano, Seung-Hee Jo, Jun Dai, Spiro Getsios, Piotr Dziunycz, Günther F.L. Hofbauer, Kara Cerveny, Giovanna Chiorino, Karine Lefort, G. Paolo Dotto

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Figure 7

Comparative gene expression and mutation profiles of skin, H/N, and lung SCCs in relation to ER signaling and/or patients’ sex.

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Comparative gene expression and mutation profiles of skin, H/N, and lung...
(A) Expression profiles of ERβ-controlled genes in clinically occurring SCCs versus normal tissues. Microarray analysis of gene expression in HKCs plus/minus ERβ-KD identified genes under ERβ control (>1.5-fold change; Supplemental Table 3). Expression profiles of these genes were examined in clinically occurring SCCs, utilizing our own and published data sets (Supplemental Table 4: Oncomine). Dark/light green and red columns on the right refer to the set of down- and upmodulated genes in HKCs with ERβ silencing that were found to be concordantly (dark colors) versus discordantly (light colors) regulated in clinically occurring SCCs in parallel with a differential gene family distribution (Supplemental Table 4). (B) Numbers of genes with mutations frequencies of 10% or more in lung and H/N SCCs from patients of one or both sexes (Fisher’s exact test < 0.05). Overlapping circles show numbers of genes with similar mutation frequency distribution in lung and H/N SCCs. For complete mutation gene list and analysis, see Supplemental Table 5. (C) Expression profiles of genes differentially expressed in female versus male H/N patients. Gene families with statistically significant enrichment are indicated on the left (Supplemental Table 6). Genes with a role in squamous differentiation are zoomed in. Log2 expression values, median centered and divided by SD, are represented.