Science Inventory

SIX1 oncoprotein is necessary for abnormal uterine basal cell development in mice exposed neonatally to DES

Citation:

Wood, C., W. Jefferson, C. Williams, AND A. Suen. SIX1 oncoprotein is necessary for abnormal uterine basal cell development in mice exposed neonatally to DES. Advances in Andrology Symposium, RTP, North Carolina, October 13, 2017.

Impact/Purpose:

Early-life exposure to estrogenic chemicals in the environment has been associated with increased susceptibility to cancer and other adverse reproductive health outcomes later in life. Biological pathways driving these effects are still largely unknown. This case study investigated the role of a developmental protein called SIX1 as a molecular driver and biomarker of latent estrogenic effects in a mouse model of early-life estrogen exposure. The ultimate goal of this work is to enable more rapid identification of AOPs for cancer incorporating life stage susceptibility.

Description:

In a classical model of latent hormonal carcinogenesis, exposing female mice on neonatal days 1-5 to the synthetic estrogen diethylstilbestrol (DES; 1 mg/kg/day) results in high incidence of uterine carcinoma. However, the biological mechanisms driving DES-induced carcinogenesis remain unclear. We previously showed that the sine oculis homeobox homolog 1 (SIX1) oncoprotein was aberrantly expressed in the uteri of mice exposed neonatally to DES. Abnormal SIX1 expression persisted with age and localized to abnormal uterine basal cells within all neoplastic lesions. In the current study, we investigate whether SIX1 is necessary for uterine basal cell differentiation and carcinogenesis in mice exposed neonatally to DES. We used a conditional knockout mouse model in which floxed Six1 is excised in the uterus using the PgR-cre transgene. We confirmed that SIX1 is effectively deleted in this model and is not induced by neonatal DES exposure by real time RT-PCR and immunoblotting immediately following treatment (day 5) and in adults (6 months). As expected, development of the female reproductive tract was normal following uterine deletion of SIX1. At 6 months of age, uterine carcinoma was observed in 4/10 (40%) of DES-exposed wildtype (Six1+/+) mice and 7/9 (78%) of DES-exposed SIX1 conditional knockout (Six1d/d) mice but not in either of the control groups. The presence of atypical hyperplasia and carcinomas in the absence of SIX1 indicates that SIX1 is not required for DES-induced carcinogenesis in the uterus. Despite this finding, we found two distinct differences between DES exposed Six1+/+ and Six1d/d mice. Abnormal basal cells expressing the basal cell marker, Keratin 14 (K14) were present throughout the entire uterus (body and horns) of DES-exposed Six1+/+ mice; however, K14 positive basal cells were restricted to the uterine body of the DES-exposed Six1d/d mice and not found in the uterine horns. Additionally, basal cell and squamous metaplasia was significantly reduced in DES-exposed Six1d/d mice, with >10-fold fewer K14 positive basal cells in endometrial glands of Six1d/d mice suggesting SIX1 is necessary for abnormal basal cell differentiation. Luminal epithelial cell number was not different between DES-exposed Six1+/+ and Six1d/d mice as indicated by the epithelial cell marker, K18. To further characterize this model, we performed a microarray analysis of all four groups at 6 months of age. There were 635 significantly different genes (>1.5 fold) between DES-exposed uteri from Six1+/+ and Six1d/d mice suggesting aberrant SIX1 expression acts as a transcription factor in this model and contributes to altered gene expression. Taken together, these data indicate that SIX1 expression is not required for DES-induced carcinogenesis in the uterus but rather serves as a key differentiation factor for aberrant basal-type progenitor cells. This abstract does not reflect U.S. EPA policy.

Record Details:

Record Type: DOCUMENT (PRESENTATION/POSTER)
Product Published Date: 10/13/2017
Record Last Revised: 06/15/2018
OMB Category: Other
Record ID: 341180

Organization:

U.S. ENVIRONMENTAL PROTECTION AGENCY

OFFICE OF RESEARCH AND DEVELOPMENT

NATIONAL HEALTH AND ENVIRONMENTAL EFFECTS RESEARCH LABORATORY

INTEGRATED SYSTEMS TOXICOLOGY DIVISION

CARCINOGENESIS BRANCH