RNA sequencing of mammary tumors induced in MTB-IGFIR transgenic mice fed 20% ISP, 1% ISP, or 20% Casein
Ontology highlight
ABSTRACT: Epidemiologic studies demonstrate that women from cultures that consume high levels of dietary soy have reduced breast cancer rates compared to women from cultures where soy consumption is typically much lower. The types of soy products consumed can also differ with Asian cultures consuming primarily minimally refined soy products while Western cultures often consume more highly refined soy products such as isolated soy protein (ISP). Our previous work showed that lifetime exposure to a diet containing 20% ISP promoted mammary tumor development in MTB-IGFIR transgenic mice. In this study, lifetime exposure to lower levels of ISP were evaluated (5% ISP and 1% ISP) to determine whether more moderate levels of ISP could protect against mammary tumorigenesis. A standard rodent diet, Teklad 2018 was also included in this study and Teklad 2018 contains a less refined form of soy, namely soybean meal. MTB-IGFIR mice fed ISP diets, independent of the concentration, displayed increased mammary tumor incidence and reduced tumor latency compared to MTB-IGFIR mice fed a 20% casein diet. Unexpectedly, MTB-IGFIR mice fed Teklad 2018 were completely protected against mammary tumor development. Although RNA sequencing of mammary tumors from ISP or casein fed mice did not identified gene expression patterns associated with the ISP diets, the ISP diets consistently promoted the expression of contractile related proteins in pubertal mammary glands. Therefore, lifetime exposure to ISP may alter gene expression in pubertal mammary glands rendering them more susceptible to transformation. Based on these findings women may want to avoid highly refined soy products such as ISP and switch to less refined forms of dietary soy until additional studies can be performed.
Project description:Epidemiologic studies demonstrate that women from cultures that consume high levels of dietary soy have reduced breast cancer rates compared to women from cultures where soy consumption is typically much lower. The types of soy products consumed can also differ with Asian cultures consuming primarily minimally refined soy products while Western cultures often consume more highly refined soy products such as isolated soy protein (ISP). Our previous work showed that lifetime exposure to a diet containing 20% ISP promoted mammary tumor development in MTB-IGFIR transgenic mice. In this study, lifetime exposure to lower levels of ISP were evaluated (5% ISP and 1% ISP) to determine whether more moderate levels of ISP could protect against mammary tumorigenesis. A standard rodent diet, Teklad 2018 was also included in this study and Teklad 2018 contains a less refined form of soy, namely soybean meal. MTB-IGFIR mice fed ISP diets, independent of the concentration, displayed increased mammary tumor incidence and reduced tumor latency compared to MTB-IGFIR mice fed a 20% casein diet. Unexpectedly, MTB-IGFIR mice fed Teklad 2018 were completely protected against mammary tumor development. Although RNA sequencing of mammary tumors from ISP or casein fed mice did not identified gene expression patterns associated with the ISP diets, the ISP diets consistently promoted the expression of contractile related proteins in pubertal mammary glands. Therefore, lifetime exposure to ISP may alter gene expression in pubertal mammary glands rendering them more susceptible to transformation. Based on these findings women may want to avoid highly refined soy products such as ISP and switch to less refined forms of dietary soy until additional studies can be performed.
Project description:RNA was isolated from mammary glands from 55 day old control mice, mice overexpressing the miR-200b/200a/429 cluster in mammary epithelial cells (MTB-200ba429) mice overexpressing the IGF-IR transgene in mammary epithelial cells (MTB-IGFIR), and mice overexpressing both the miR-200b/200a/429 cluster and the IGF-IR transgene in mammary epithelial cells (MTB-IGFIRba429)
Project description:The role of diet in the prevention of breast cancer is widely accepted, yet little is known on how early dietary effects mitigate adult cancer risk. Soy consumption is associated with reduced breast cancer risk in women, an effect largely attributed to the soy isoflavone genistein (GEN). We previously showed lower chemically-induced mammary tumor incidence in young adult rats with lifetime dietary intake of soy protein isolate (SPI), a highly refined soy product in infant formula, than in those fed the control diet Casein (CAS). To gain insight into signaling pathways underlying dietary tumor protection, we performed genome-wide expression profiling of mammary epithelial cells from young adult rats lifetime fed CAS, SPI, or supplemental GEN-based diets. We identified mammary epithelial genes regulated by SPI (79 total) and GEN (99 total) using Affymetrix rat 230A GeneChip arrays and found minimal overlap in gene expression patterns. We showed that the regulated transcripts functionally cluster in biochemical pathways involving metabolism, immune response, signal transduction, and ion transport. We confirmed the differential expression of Wnt (Wnt5a, Sfrp2) and Notch (Notch2, Hes1) signaling components by SPI and/or GEN using QPCR. Wnt pathway inhibition by GEN was supported by lower Cyclin D1 immunoreactivity in mammary ductal epithelium of GEN relative to CAS and SPI, despite their comparable levels of membrane-localized E-cadherin and β-catenin. Identification of distinct GEN and SPI responsive genes in mammary epithelial cells may define early events contributing to tumor protection by diet relevant to the prevention of breast and other types of cancer. Keywords: Comarative genomic hybridization
Project description:Molecular profiling was used to classify mammary tumors that develop in MTB-IGFIR transgenic mice. It was determined that the primary mammary tumors (PMT), which develop due to elevated expression of the type I insulin-like growth factor receptor (IGF-IR) in mammary epithelial cells, most closely resemble murine tumors with basal-like or mixed gene expression profiles and with human basal-like breast cancers. Downregulation of IGF-IR transgene in MTB-IGFIR tumor-bearing mice leads to the regression of most of the tumors followed by tumor re-appearance in some of the mice. These tumors that re-appear following IGF-IR transgene downregulation do not express the IGF-IR transgene and cluster with murine mammary tumors that express a mesenchymal gene expression profile and with human claudin-low breast cancers. Therefore, IGF-IR overexpression in murine mammary epithelial cells induces mammary tumors with primarily basal-like characteristics while tumors that develop following IGF-IR downregulation express a gene signature that most closely resembles human claudin-low breast tumors. Three conditions: 8 wild type (WT) mammary glands, 11 primary mammary tumor (PMT) samples, 9 recurrent spindle tumor (RST) samples, each sample was hybridized against a universal mouse reference RNA
Project description:We have previously shown that transgenic overexpression of the miR-200b/200a/429 cluster prevents mammary tumor development in MTB-IGFIR mice. In this study we evaluated whether the miR-200b/200a/429 cluster could also prevent mammary tumor development from a different oncogene, namely Neu/Erbb2. We found that transgenic overexpression of Neu/Erbb2 in MTB-TAN mice induce rapid mammary tumor development and co-overexpression of the miR-200b/200a/429 cluster with Neu/Erbb2 completely prevent mammary epithelial transformation and tumor development
Project description:The role of diet in the prevention of breast cancer is widely accepted, yet little is known on how early dietary effects mitigate adult cancer risk. Soy consumption is associated with reduced breast cancer risk in women, an effect largely attributed to the soy isoflavone genistein (GEN). We previously showed lower chemically-induced mammary tumor incidence in young adult rats with lifetime dietary intake of soy protein isolate (SPI), a highly refined soy product in infant formula, than in those fed the control diet Casein (CAS). To gain insight into signaling pathways underlying dietary tumor protection, we performed genome-wide expression profiling of mammary epithelial cells from young adult rats lifetime fed CAS, SPI, or supplemental GEN-based diets. We identified mammary epithelial genes regulated by SPI (79 total) and GEN (99 total) using Affymetrix rat 230A GeneChip arrays and found minimal overlap in gene expression patterns. We showed that the regulated transcripts functionally cluster in biochemical pathways involving metabolism, immune response, signal transduction, and ion transport. We confirmed the differential expression of Wnt (Wnt5a, Sfrp2) and Notch (Notch2, Hes1) signaling components by SPI and/or GEN using QPCR. Wnt pathway inhibition by GEN was supported by lower Cyclin D1 immunoreactivity in mammary ductal epithelium of GEN relative to CAS and SPI, despite their comparable levels of membrane-localized E-cadherin and β-catenin. Identification of distinct GEN and SPI responsive genes in mammary epithelial cells may define early events contributing to tumor protection by diet relevant to the prevention of breast and other types of cancer. Experiment Overall Design: Female Sprague-Dawley rats, fed one of the three purified diets, were studied at postnatal day50. Total RNA( each RNA samples were extracted from #4 mammary gland from two animals under the same diet group) purified from non-tumor tissue within the proximal half of each colon, was used to prepare biotinylated probes, which were hybridized to Affymetrix RAE230 rat microarrays.
Project description:Molecular profiling was used to classify mammary tumors that develop in MTB-IGFIR transgenic mice. It was determined that the primary mammary tumors (PMT), which develop due to elevated expression of the type I insulin-like growth factor receptor (IGF-IR) in mammary epithelial cells, most closely resemble murine tumors with basal-like or mixed gene expression profiles and with human basal-like breast cancers. Downregulation of IGF-IR transgene in MTB-IGFIR tumor-bearing mice leads to the regression of most of the tumors followed by tumor re-appearance in some of the mice. These tumors that re-appear following IGF-IR transgene downregulation do not express the IGF-IR transgene and cluster with murine mammary tumors that express a mesenchymal gene expression profile and with human claudin-low breast cancers. Therefore, IGF-IR overexpression in murine mammary epithelial cells induces mammary tumors with primarily basal-like characteristics while tumors that develop following IGF-IR downregulation express a gene signature that most closely resembles human claudin-low breast tumors.
Project description:The linkage between nutrition and cancer prevention is an intriguing concept that is gaining widespread support based on epidemiological and animal studies. Multiple mechanisms likely underlie dietary protection against cancer, with effects influenced by target tissue response, cell-cell interactions and developmental context. Given the negative correlation between breast cancer incidence and intake of soy foods by Asian women, and the increasing consumption of soy protein-based formula by infants in the Western world, we have studied soy protein isolate (SPI) used in most infant formula as a paradigm to evaluate diet as a risk factor in a rodent model of mammary cancer. We previously demonstrated that lifetime exposure to dietary SPI reduced the incidence of N-methyl-N-nitrosourea-induced mammary tumors in young adult rats relative to those fed the control diet Casein (CAS). This protection was associated with increased tumor suppressor PTEN and decreased Wnt signaling component expression in mammary epithelial cells at postnatal day (PND) 50 prior to carcinogen insult. To identify early events contributing to mammary tumor suppression by diet, we used Affymetrix RAE230A GeneChips containing 14280 probe sets and the GeneSpring Robust Multi-array program to analyze genomic profiles of mammary glands of prepubertal (PND21) rats lifetime exposed to SPI or CAS. Experiment Overall Design: Female SD rats are exposed to either casein/soy protein isolate based AIN-93G diet since gestation day 4 till weaning. Whole mammary gland were harvested for the gene analysis by microarray.