Abstract 3283: Conditional Low-molecular-weight (LMW) cyclin E-induced mammary tumorigenesis

Abstract

Abstract Cyclin E overexpression and generation of hyperactive LMW cyclin E is oncogenic and has been implicated in a variety of human cancers. We have developed a doxycycline-inducible LMW cyclin E transgenic mouse model to determine the dependence of established mammary adenocarcinomas on continued expression of LMW cyclin E. Mice harboring this TetO-LMW-E transgene, referred to as TLMW, were mated to MMTV-rtTA transgenic mice (MTB) to yield bitransgenic MTB/TLMW mice. Morphological examination of hematoxylin and eosin-stained sections showed that the mammary glands of induced MTB/TLMW mice following four days of doxycycline induction were hyperplastic compared with uninduced MTB/ TLMW animals. Consistent with this, immunohistochemistry using antibodies specific for BrdU revealed a 2-fold increase in BrdU incorporation in the mammary epithelium of induced bitransgenic females compared with uninduced mice (17.1+/−5.3% versus 30.9+/−4.9%, P=0.03). This data indicate that LMW cyclin E overexpression in the mammary epithelium increased proliferation. To determine whether sustained induction of transgene expression would result in mammary tumorigenesis in MTB/TLMW mice, a cohort of 27 bitransgenic females was treated with doxycycline beginning at 6 weeks of age. 41% of these mice developed tumors within 2 years of starting doxycycline with kinetics (T1/2= 20 months) resembling that described for MMTV-LMW cyclin E mice. Similarly, the predominant type of tumor observed for the inducible LMW forms was glandular adenocarcinomas with adenosquamous differentiation. Lung metastases are known to arise in a fraction of MMTV-LMW cyclin E transgenic mice harboring primary mammary tumors. Accordingly, we sacrificed a subset of mammary tumor bearing MTB/TLMW mice and examined organs at necropsy for evidence of metastatic spread. 4 of 7 mice examined had lung metastasizes (57%). Control mice without dox never developed mammary tumors. To shed further light on the molecular mechanisms of mammary tumorigenesis caused by LMW-E expression, we conducted DNA microarray analyses. Using the R package limma, 641 probes out of 30,854 probes were identified as differentially regulated by at least 2-fold by LMW-E in the tumors when compared to the contralateral mammary glands. Gene Ontology analysis identified genes on this list that were involved in a variety of processes, including cell cycle control of chromosome replication, and DNA methylation and transcriptional repression signaling. Based on histology and gene expression profile of the resultant mammary tumors, we propose that the MTB/TLMW is a murine model of breast cancer resembling the human invasive basal/triple negative breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3283. doi:1538-7445.AM2012-3283