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Identification of Wnt responsive genes using a murine mammary epithelial cell line model system

Taneyhill, Lisa and Pennica, Diane (2004) Identification of Wnt responsive genes using a murine mammary epithelial cell line model system. BMC Developmental Biology, 4 (6). pp. 1-14. ISSN 1471-213X.

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Background: The Wnt/Wg pathway plays an important role in the developmental program of many cells and tissues in a variety of organisms. In addition, many Wnts and components of their downstream signaling pathways, such as β-catenin and APC, have been implicated in tumorigenesis. Over the past years, several genes have been identified as Wnt responsive, including c-myc, siamois, and cyclin D1. Results: In order to identify additional genes responsive to Wnt signaling that contribute to the transformed phenotype, we performed a cDNA subtractive hybridization screen between a mouse mammary epithelial cell line that overexpresses Wnt-1 (C57MG/Wnt-1) and the parental cell line (C57MG). The screen identified a total of 67 genes to be up-regulated in response to Wnt signaling. Of these 67 genes, the up-regulation of 62 was subsequently confirmed by Northern and dot blot analyses (and, for a subset, semi-quantitative PCR) of RNA isolated from C57MG cells subjected to (1) an independent Wnt-1 retroviral infection, and (2) co-culture with Wnt-1 expressing cells. Among the confirmed Wnt-1 responsive genes, we further characterized a mouse homolog of the human transcription factor Basic Transcription Element Binding protein 2 (BTEB2), Wnt-1 Responsive Cdc42 homolog (Wrch-1), and Wnt-1 Induced Secreted Protein (WISP-1). Conclusion: Several novel genes were identified in this screen, as well as others that have been shown previously to be regulated by Wnt signaling, such as connexin43. The results indicate that cDNA subtractive hybridization is a useful method for identifying genes involved in the process of Wnt-1-induced transformation.

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Additional Information:Received: 18 December 2003; Accepted: 12 May 2004; Published: 12 May 2004. BMC Developmental Biology 2004, 4:6 This article is available from: © 2004 Taneyhill and Pennica; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. Authors' contributions: LT carried out all of the molecular studies and experiments described herein, with the exception of the actual screen process, sequencing, and QRT-PCR experiments that were performed by DP. LT, in conjunction with Dr. Arnold J. Levine, conceived of the study and participated in its design and coordination. All authors read and approved the final manuscript. Acknowledgments: This work was supported by the NIH Cancer Training Grant T32 CA-09528. L. Taneyhill was supported in part by a fellowship from the New Jersey Commission on Cancer Research (98-2001-CCR-00). The authors would like to especially thank Dr. Arnold J. Levine for his support and encouragement of this work. Finally, the authors would like to thank Dr. Marianne Bronner-Fraser for her editorial assistance and helpful suggestions.
Issue or Number:6
Record Number:CaltechAUTHORS:TANbmcdb04
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:267
Deposited By: Archive Administrator
Deposited On:13 May 2005
Last Modified:02 Oct 2019 22:31

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