SLUG/SNAI2 and Tumor Necrosis Factor Generate Breast Cells With CD44+/CD24- Phenotype

<p>Abstract</p> <p>Background</p> <p>Breast cancer cells with CD44+/CD24- cell surface marker expression profile are proposed as cancer stem cells (CSCs). Normal breast epithelial cells that are CD44+/CD24- express higher levels of stem/progenitor cell associated genes. We, amongst others, have shown that cancer cells that have undergone epithelial to mesenchymal transition (EMT) display the CD44+/CD24- phenotype. However, whether all genes that induce EMT confer the CD44+/CD24- phenotype is unknown. We hypothesized that only a subset of genes associated with EMT generates CD44+/CD24- cells.</p> <p>Methods</p> <p>MCF-10A breast epithelial cells, a subpopulation of which spontaneously acquire the CD44+/CD24- phenotype, were used to identify genes that are differentially expressed in CD44+/CD24- and CD44-/CD24+ cells. Ingenuity pathway analysis was performed to identify signaling networks that linked differentially expressed genes. Two EMT-associated genes elevated in CD44+/CD24- cells, SLUG and Gli-2, were overexpressed in the CD44-/CD24+ subpopulation of MCF-10A cells and MCF-7 cells, which are CD44-/CD24+. Flow cytometry and mammosphere assays were used to assess cell surface markers and stem cell-like properties, respectively.</p> <p>Results</p> <p>Two thousand thirty five genes were differentially expressed (p < 0.001, fold change ≥ 2) between the CD44+/CD24- and CD44-/CD24+ subpopulations of MCF-10A. Thirty-two EMT-associated genes including SLUG, Gli-2, ZEB-1, and ZEB-2 were expressed at higher levels in CD44+/CD24- cells. These EMT-associated genes participate in signaling networks comprising TGFβ, NF-κB, and human chorionic gonadotropin. Treatment with tumor necrosis factor (TNF), which induces NF-κB and represses E-cadherin, or overexpression of SLUG in CD44-/CD24+ MCF-10A cells, gave rise to a subpopulation of CD44+/CD24- cells. Overexpression of constitutively active p65 subunit of NF-κB in MCF-10A resulted in a dramatic shift to the CD44+/CD24+ phenotype. SLUG overexpression in MCF-7 cells generated CD44+/CD24+ cells with enhanced mammosphere forming ability. In contrast, Gli-2 failed to alter CD44 and CD24 expression.</p> <p>Conclusions</p> <p>EMT-mediated generation of CD44+/CD24- or CD44+/CD24+ cells depends on the genes that induce or are associated with EMT. Our studies reveal a role for TNF in altering the phenotype of breast CSC. Additionally, the CD44+/CD24+ phenotype, in the context of SLUG overexpression, can be associated with breast CSC "stemness" behavior based on mammosphere forming ability.</p

Key signalling nodes in mammary gland development and cancer. The Snail1-Twist1 conspiracy in malignant breast cancer progression

Breast cancer is the most common cancer among women, and despite significant advances in diagnosing and treating it, metastatic spread of cancer cells results in a high mortality rate. Epithelial-to-mesenchymal transition (EMT) is an embryonic program in which epithelial cells lose their characteristics and gain mesenchymal features. Therefore, EMT might play a very important role during malignant tumour progression. In this review we summarise recent advances in breast cancer research with a particular focus on the transcription factors Snail1 and Twist1. Besides discussing the role of EMT in normal mammary gland development, we describe regulatory mechanisms involving newly discovered upstream regulators and microRNAs, the association of EMT with breast cancer stem cells, and the involvement of the tumour microenvironment in breast cancer progression

The Pan-STARRS1 Surveys

Pan-STARRS1 has carried out a set of distinct synoptic imaging sky surveys including the $3\pi$ Steradian Survey and the Medium Deep Survey in 5 bands ($grizy_{P1}$). The mean 5$\sigma$ point source limiting sensitivities in the stacked 3$\pi$ Steradian Survey in $grizy_{P1}$ are (23.3, 23.2, 23.1, 22.3, 21.4) respectively. The upper bound on the systematic uncertainty in the photometric calibration across the sky is 7-12 millimag depending on the bandpass. The systematic uncertainty of the astrometric calibration using the Gaia frame comes from a comparison of the results with Gaia: the standard deviation of the mean and median residuals ($\Delta ra, \Delta dec$) are (2.3, 1.7) milliarcsec, and (3.1, 4.8) milliarcsec respectively. The Pan-STARRS system and the design of the PS1 surveys are described and an overview of the resulting image and catalog data products and their basic characteristics are described together with a summary of important results. The images, reduced data products, and derived data products from the Pan-STARRS1 surveys are available to the community from the Mikulski Archive for Space Telescopes (MAST) at STScI

The Pan-STARRS1 Surveys

Pan-STARRS1 has carried out a set of distinct synoptic imaging sky surveys including the $3\pi$ Steradian Survey and the Medium Deep Survey in 5 bands ($grizy_{P1}$). The mean 5$\sigma$ point source limiting sensitivities in the stacked 3$\pi$ Steradian Survey in $grizy_{P1}$ are (23.3, 23.2, 23.1, 22.3, 21.4) respectively. The upper bound on the systematic uncertainty in the photometric calibration across the sky is 7-12 millimag depending on the bandpass. The systematic uncertainty of the astrometric calibration using the Gaia frame comes from a comparison of the results with Gaia: the standard deviation of the mean and median residuals ($\Delta ra, \Delta dec$) are (2.3, 1.7) milliarcsec, and (3.1, 4.8) milliarcsec respectively. The Pan-STARRS system and the design of the PS1 surveys are described and an overview of the resulting image and catalog data products and their basic characteristics are described together with a summary of important results. The images, reduced data products, and derived data products from the Pan-STARRS1 surveys are available to the community from the Mikulski Archive for Space Telescopes (MAST) at STScI