bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3310: microRNAs regulate the transition between EMT and MET breast cancer stem cell states
Authors: Liu Suling , Shang Li , Bai Shoumin , Martin-Trevino Rachel , McDermott Sean , Ginestier Christophe , Ibarra Ingrid , Baker Trenton , Malik Fayaz , Ke Jia , Patel Shivani H. , Luther Tahra , Korkaya Hasan , Zhang Kevin J. , Clouthier Shawn G. , Hannon Gregory J. , Wicha Max S. 
Published: August 2012

Abstract
Recent studies have suggested similarities between cancer stem cells and the epithelial mesenchymal transition (EMT) state. In contrast, other studies suggest that these “states” are mutually exclusive. Our studies suggest that these divergent views may be explained by the existence of multiple stem cell states, which are regulated by microRNAs. Utilizing primary breast tissue and established cell lines, we demonstrate that both normal and malignant breast stem cells exist in distinct, inter-convertible states. The EMT-like state is characterized by expression of vimentin and N-cadherin, slug, snail and twist transcription factors. EMT-like CSCs have a mesenchymal morphology, are largely quiescent, invasive and characterized by expression of the CSC markers CD44+CD24− and are EpCAM−CD49f+. In contrast, the MET (mesenchymal epithelial transition) state of CSCs is characterized by an epithelial morphology and expression of E-cadherin and EpCAM. MET-like CSCs undergo self-renewal and express the CSC marker Aldehyde dehydrogenase (ALDH) and are EpCAM+CD49F+. A subpopulation of cells expressing both CD44+CD24− and ALDH may represent cells in transition between these states. This transition is regulated by signals in the microenvironment which in turn modulate microRNA networks. Expression induction of mir100 in MCF10A cells and several cancer cell lines resulted in a decrease of ALDH-positive CSC population with a concomitant increase in the CD24−CD44+ population accompanied by induction of EMT. We demonstrated mir100 effects are mediated by targeting BMPR2, SMARCA5 and SMARCD1, all of which may contribute to induction of EMT. Moreover, we show that mir100 overexpression induces cellular quiescence as shown by Ki67 and Brdu staining. Induction of mir100 expression immediately upon orthotopic implantation or after the tumors are established significantly reduced the subsequent tumor growth in NOD/SCID mice. In contrast of overexpression of mir100 stimulates cell invasion in vitro matrigel assay. The existence of multiple cancer stem cell states has important implications for understanding stem cell plasticity as well as tumor growth and metastasis. In addition, the existence of these states has implications for the development of CSC-targeting therapeutics.
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 3310. doi:1538-7445.AM2012-3310
Abstract 3312: Targeting Notch signaling to abrogate the increase in breast cancer stem cells induced by hypoxia
Authors: Conley Sarah J. , Gheordunescu Elizabeth , Theisen Rebecca , Wicha Max 
Published: August 2012

Abstract
There is increasing evidence from our lab and others supporting the “cancer stem cell” hypothesis in breast cancer, which deems that tumors are driven a small population of cells exhibiting stem cell properties. These cancer stem/progenitor cells may also be responsible for mediating tumor metastasis and resistance to cancer treatments. Recently, several new cancer therapies targeting tumor angiogenesis have been developed and validated in clinical trials. However, clinical practice has revealed that these therapies ultimately result in drug resistance and do not prolong patient survival by more than a few months. It has been reported that these angiogenesis inhibitors promote tumor invasiveness and metastasis in rodent breast cancer models. We have used in vitro and in vivo assays to study the role of hypoxia in regulating mammary cancer stem cells. We have shown that breast tumor stem cell population increases in response to hypoxia in vitro and in tumors from anti-angiogenic treated mice. We hypothesize that this leads to increased tumor aggressiveness and may explain the resistance to these drugs seen in patients. In vivo, both sunitinib and bevacizumab treatment increased intratumoral hypoxia as well as the percentage of cells expressing ALDH as assessed by the Aldefluor assay in SUM-159 and MDA-MB-231 breast cancer xenografts in NOD/SCID mice. In addition, tumor cells obtained from sunitinib treated mice exhibit enhanced growth of secondary tumors in NOD/SCID mice, further supporting an increase in the tumor stem cell population. We also demonstrate that hypoxia (1-2% oxygen) enriches for stem cells by at least 2 fold in several breast cancer cell lines grown in vitro. Utilizing siRNA, we identified that HIF1a is required for the increased cancer stem cell population following hypoxia treatment. We next tested whether a gamma secretase inhibitor (GSI) can block hypoxia's ability to increase the cancer stem cell population since HIF1a is reported to activate the Notch signaling pathway, and this pathway is known to regulate stem cell self renewal. Treatment of cells with the GSI completely abrogated the increase in ALDH+ cells under hypoxic conditions. We are now testing the ability of GSIs to block the increase in CSCs seen in mice treated with anti-angiogenic drugs. This study may reveal that therapies targeting tumor angiogenesis should be combined with GSI treatments targeting the cancer stem cell population.
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 3312. doi:1538-7445.AM2012-3312
Abstract 3313: Novel roles of CCN6 in mesenchymal-epithelial transition (MET) and loss of stem cell properties of breast cancer cells
Authors: Huang Wei , Kleer Celina G. 
Published: August 2012

Abstract
Background: Epithelial-mesenchymal transition (EMT) and its reverse process, mesenchymal-epithelial transition (MET), play key roles in regulating embryogenesis. Recent studies have shown that aberrant EMT activation contributes to cancer progression and metastasis. Links between EMT and the gain of stem cell properties have been suggested in physiological and pathological situations. CCN family protein 6 (CCN6/WISP3) is a novel tumor suppressor that regulates diverse cellular functions. Our previous studies have shown the blockade of CCN6 by short hairpin RNA triggers EMT program in benign breast epithelial cells, with decreased expression of epithelial marker E-cadherin and increased activation of corresponding transcriptional repressor, Zeb1 and Snail. Moreover, loss of CCN6 expression also endows benign breast epithelial cells with growth factor-independent properties, such as cell survival and anoikis resistance. However, little is known regarding the functions of CCN6 overexpression in breast cancer progression. Methods: We overexpressed CCN6 in breast cancer cell lines, MDA-MB-231 and SUM-159, which are aggressive, have mesenchymal-like features and low levels of endogenous CCN6. CCN6 provoked functions were studied in vitro by proliferation, motility and invasion assays. Aldehyde dehydrogenase (ALDH1) was used to identify and investigate CCN6 roles in human breast cancer stem cells. Results: We showed that overexpression of CCN6 inhibits cell proliferation, motility and invasion. Importantly, CCN6 overexpression in these cells led to phenotypic changes towards mesenchymal-epithelial transition (MET), characterized by loss of mesenchymal cell marker Vimentin and downregulation of transcriptional factor, Zeb1. CCN6 overexpression significantly reduced the stem cell population of MDA-MB-231 and SUM-159 breast cancer cells. Conclusions: Our data suggest a novel function of CCN6 in regulating MET and reducing the stem cell population of aggressive breast cancer cells.
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 3313. doi:1538-7445.AM2012-3313
Abstract 3315: Malignant melanoma cells reciprocate aspects of the embryonic neural crest migration program to promote invasion and increased plasticity
Authors: Bailey Caleb M. , Morrison Jason A. , Kulesa Paul M. 
Published: August 2012

Abstract
Melanocytes derive from a highly invasive, multipotent embryonic cell population known as the neural crest. It has been proposed that this ancestral relationship predisposes melanocytes to having metastatic characteristics following neoplastic transformation to melanoma. In support of this hypothesis, we have previously shown that aggressive melanoma cells transplanted into the chick embryonic neural crest microenvironment respond to neural crest signaling mechanisms by emigrating along defined host neural crest pathways. To evaluate whether melanoma cells usurp aspects of the neural crest program typically silenced in differentiated melanocytes, we have employed the chick embryo transplant model coupled with laser capture microdissection to perform an in vivo molecular analysis of more than 70 neural-crest-related genes. Transplanted cells included c8161 aggressive melanoma cells, a poorly aggressive isogenic counterpart termed c81-61, and primary human melanocytes. We demonstrate that all three cell types express many neural-crest-related genes, but that aggressive cells aberrantly express some genes typically lost during melanocyte differentiation. Furthermore, the analysis revealed a significant increase in the ability of aggressive melanoma cells to respond to changing microenvironments by altering gene expression signatures, while transplanted melanocytes appeared to actively suppress these pathways. This increased responsiveness may be attributed to the dynamic regulation of Eph receptor tyrosine kinases and their cognate ephrin ligands, many of which were differentially expressed in aggressive melanoma cells versus poorly aggressive melanoma or primary melanocytes. Thus, metastatic melanoma cells usurp portions of the neural crest program to promote increased invasiveness and plasticity.
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 3315. doi:1538-7445.AM2012-3315
Abstract 3317: The roles of TMCC3 in breast cancer cells
Authors: Wang Ya- Hui , Yu Alice L. 
Published: August 2012

Abstract
There is growing evidence showing that cancer stem cells are more likely to metastasize and are more frequently detected in the circulation. Thus, it will be important to understand the molecular characteristics of breast cancer stem cells (BCSCs) which may lead to the development of novel targets for CSC-directed therapy. Through phosphoproteomic analysis of BCSC and non-BCSC, we identified differential expression of the transmembrane and coiled-coil domain family 3 (TMCC3). TMCC3 belongs to TEX28 family and is predicted to be an integral membrane protein. The function of TMCC3 is unknown. We found higher expression of TMCC3 in invasive breast cancer cell line and BCSCs than less-invasive breast cancer cell lines, and non-BCSCs, respectively. In addition, TMCC3 expression was greater in distant lymph node metastasis than in primary tumor of human breast cancer xenograft in mouse. Silencing of TMCC3 led to G1 phase arrest with increased apoptotic cells, along with decreased mammosphere formation and ALDH activity which are important features of cancer stem cell. In our previous investigation, the IGF-1R/PI3K/Akt/mTOR pathway was shown to be important for BCSCs survival and maintenance. To explore the possible involvement of TMCC3 in this pathway, we further examined the Akt and mTOR activation in TMCC3-slienced BCSCs. We found the Akt phosphorylation was lost in BCSCs upon TMCC3 knocked down. These findings suggest that TMCC3 is essential for breast cancer cell survival, self-renewal and metastasis. Thus, inhibition of TMCC3 may provide a new therapeutic strategy targeting stem cells.
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 3317. doi:1538-7445.AM2012-3317
Abstract 332: The role of OLA1, a novel obg-like ATPase, in epithelial-mesenchymal transition
Authors: Bai Li , Rubio Valentina , Mao Renfang , Chen Huarong , Mansour Oula C. , Shi Zheng-Zheng 
Published: August 2012

Abstract
Human OLA1 is a 45 kDa cytosolic protein that belongs to the YchF subfamily of Obg-like P-loop GTPases and is ubiquitously expressed in most tissues. YchF proteins are highly conserved from yeast to humans and all have a greater affinity for ATP than GTP. Based on structure analysis, OLA1 is predicted to be a regulatory protein that interacts with its client protein(s) and exerts its downstream functions by the conformational switch between the ADP- and ATP-bound forms. However, the actual physiological functions of OLA1 are poorly understood. Our earlier studies have suggested that OLA1 is a negative regulator of cellular antioxidant response and functions through post-transcriptional mechanisms (PNAS 2009; 106:15356). Oxidative stress, mediated by reactive oxygen species (ROS), has been implicated in epithelial-mesenchymal transition (EMT), a fundamental phenotypic alteration closely associated with cancer progression and metastasis. In this study we investigated the effect of OLA1 on EMT and the possible underlying mechanisms. We first demonstrated that the human lung epithelial carcinoma A549 cells, after exposure to transforming growth factor (TGF)-β1 for 3 days, underwent EMT as evidenced by acquisition of a mesenchymal-like morphology, down-regulation of epithelial-specific marker E-cadherin, and induction of mesenchymal marker vimentin. However, knockdown of OLA1 with sequence-specific siRNA in A549 cells resulted in a significant attenuation of the TGF-β1-induced EMT phenotypes, including those in cellular morphology and the EMT markers E-cadherin and vimentin, as compared with the cells transfected with the control non-targeting siRNA. Next, we questioned if this inhibitory effect on EMT was through inhibiting the Akt/GSK3β/Snail pathway. When treated with TGF-β1, OLA1-knockdown cells showed impaired GSK-3β phosphorylation (indicating an overactivation of GSK-3β), and subsequently reduced abundance of the transcription factor Snail that functions as the suppressor of E-cadherin gene expression. Concurrently, OLA1-knockdown cells exhibited significantly decreased TGF-β1-mediated intracellular ROS production assessed by DCFDA staining and flow cytometry analysis. Furthermore, we found that treatment of H2O2 abrogated resistance of OLA1-knockdown cells to TGF-β1-induced EMT, evidenced by phosphorylation of GSK-3β, upregulation of Snail and changes in expression of EMT markers. These results suggest that OLA1 plays an important regulatory role in EMT of human cancer cells, and inhibition of OLA1 may have a negative effect on EMT, probably through down-regulation of the ROS-dependent TGF-β1 signaling pathway.
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 332. doi:1538-7445.AM2012-332
Abstract 3322: Phenotypic plasticity in the normal breast
Authors: Sauder Candice Anne Marcum , Koziel Jillian E. , Choi MiRan , Fox Melanie J. , Badve Sunil , Blosser Rachel J. , Mathieson Theresa , Rufenbarger Constance A. , Henry Jill E. , Storniolo Anna Maria V. , Herbert Brittney-Shea , Clare Susan E. 
Published: August 2012

Abstract
Metaplasia is observed in almost all epithelial cancers. The origins of metaplasia are obscure although chronic inflammation is thought to be one etiology. We present data that suggests that an origin of metaplasia is the normal resident stem cell population. Methods: Starting from 10 gauge tissue cores of normal breast donated to the Komen Tissue Bank, 28 normal mammary epithelial (HME) and 33 normal stromal (HMS) cell lines were established using an organoid isolation method after digestion with enzymes for 24 hours. The HME cell lines were characterized by immunohistochemistry (IHC). Ploidy was assayed. Cellular morphology was observed both on two-dimensional and in three-dimensional culture systems. The HME cells were subjected to FACS analysis using multiple antibodies. Results: 96.9% of early passage cells are diploid. The HME cells express vimentin, CK 5/6, p63, CD 10, CK 18, and HER-1 when grown on two dimensional plastic surfaces. Cells placed in the center of a sandwich of Matrigel uniformly make spheres 37mm-325mm in diameter. Hematoxylin and eosin staining of the formalin-fixed and paraffin-embedded sections of these spheres reveal keratinized squamous differentiation. When the cells are grown on Laminin, Collagen Type IV, or Fibronectin surfaces multiple cell types are observed including osteoclasts, distinguished by the presence of Tartrate Resistant Acid Phospatase; and chondrocytes, confirmed by staining with Alcian Blue. Other cells with a spindle-shape and cytoplasmic vacuoles turn a dark reddish-brown color when stained with Oil Red O, characteristics of adipocytes. In other areas of the culture, the cells form a syncitium and they express the protein MyoD, a marker of immature muscle. Finally, there are numerous cells with long, dendritic processes. These cells express Nestin, glial fibrillary acidic protein (GFAP), and beta-III tubulin. Using FACS, the HME cells were found to be CD49f positive and EpCAM negative. Multiple nucleoli were confirmed using anti-Nucleostemin IHC. Conclusions: Phenotypic plasticity is common to all the HME cell lines characterized to date. Differentiation into cells of mesodermal and ectodermal origin, CD49+/EpCAM- by FACS, and the presence of multiple nucleoli suggest that the isolated cells are a multipotent/stem cell residing in the normal adult breast. These cells, through a series of yet to be elucidated events, may be the cells of origin of both benign and malignant metaplasia observed in breast lesions.
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 3322. doi:1538-7445.AM2012-3322
Abstract 3323: Hyaluronan synthase 2 promotes breast cancer bone metastasis by stimulating interaction of cancer stem-like cells with tumor-associated macrophage
Authors: Okuda Hiroshi , Xing Fei , Kobayashi Aya , Watabe Misako , Pai Sudha K. , Liu Wen , Pandey Puspa R. , Fukuda Koji , Iiizumi Megumi 
Published: August 2012

Abstract
The molecular mechanism of metastatic progression, particularly the roles of microenvironment of the metastasized organs, is as yet poorly understood. In this study, we have isolated cancer stem cell-like cells (CSC) from human breast cancer cell lines, MDA-MB231, MDA231BoM and MDA231BrM. CSC were isolated from each line using surface markers (CD24-, CD44+, ESA+) and their tumor initiating abilities were tested in animal. We then performed an analysis of global expression profile for these CSCs using the Affymetrix expression array and found that among all up-regulated genes, two genes (HAS2 and MMP1) were significantly correlated with overall- and metastasis-free survival when they were cross-examined in GEO database. To further examine the role of HAS2 in tumor metastasis in vivo, we prepared CSC from MDA231BoM with or without carrying a shRNA expression vector for HAS2. The CSC were then implanted into nude mice by intra-cardiac injection followed by oral administration of 4MU, a specific inhibitor of HAS genes. We found that the injection of CSC of MDA231BoM which expresses shRNA to HAS2 significantly improved the survival rate of animals. In addition, administration of 4MU significantly suppressed tumor metastasis of CSC, suggesting that inhibition of HAS2 can indeed reduce the incidence of metastasis in vivo. We also examined the effect of tumor associated macrophage (TAM) on growth of CSC in animals by injecting CSC of MDA231-BoM with or without TAM into the tibial bone. We found that co-injection of TAM with CSC significantly augmented the growth of tumor in their legs. Moreover, the interaction between cancer cell and TAMs enhanced secretion of PDGF-BB from TAMs which then activated stromal cells and in turn enhanced self-renewal of CSCs by secretion of FGF9 and FGF7. We also found that treatment of bone marrow stem cell (BMSC) with PDGF followed by co-culture of CSC significantly promoted proliferation of CSC. Collectively, our results suggest that TAM and BMSC promote self-renewal of CSC in bone microenvironment via direct interaction between hyaluronan and CD44. Our results also provide strong rationale to target HAS2 by using 4MU in order to block the metastatic progression of CSC in 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 3323. doi:1538-7445.AM2012-3323
Abstract 3326: ΔNp63α is a regulator of epithelial-mesenchymal transition in human keratinocytes
Authors: Oh Ju-Eun , Liu Zi , Kim Reuben , Shin Ki-Hyuk , Park No-Hee , Kang Mo K. 
Published: August 2012

Abstract
p63 is a p53 family protein required for morphogenesis and postnatal regeneration of epithelial tissues. Here we demonstrate that ΔNp63α, a p63 isoform lacking N-terminal transactivation domain, induces epithelial-mesenchymal transition (EMT) in primary human keratinocytes in TGF-β-dependent manner. Rapidly proliferating normal human epidermal keratinocytes (NHEK) were infected with retroviral vector expressing ΔNp63α, ΔNp63α, ΔNp63α, or empty vector, and serially subcultured until replicative senescence. No phenotypic changes were observed until the culture reached senescence. Then, cells transduced with ΔNp63α and ΔNp63α demonostrated morphological changes resembling mesenchymal cells, while ΔNp63α did not trigger such change in phenotype. Treatment with exogenous TGF-β accelerated EMT in pre-senescent ΔNp63α-transduced cells, and inhibition of TGF-β signaling reversed the EMT phenotype. Molecular markers of EMT, such as loss of E-Cadherin and cytokeratin 14 expression, were observed in cells expressing ΔNp63α but not those expressing ΔNp63α. TGF-β treatment alone led to growth arrest in control NHEK with no evidence of EMT, indicating that ΔNp63α altered the cellular response to TGF-β treatment. ΔNp63α-transduced cells acquiring EMT gained ability to be differentiated to osteo/odontogenic and adipogenic pathways, resembling mesenchymal stem cells (MSCs). Furthermore, these cells expressed enhanced levels of Nanog and Lin28, which are transcription factors associated with pluripotency. These data indicate that ΔNp63α triggered EMT in TGF-β-dependent manner, while ΔNp63α only caused morphologically changes resembling mesenchymal cells. The current study suggests that mesenchymal transition of normal skin keratinocytes might be a novel approach to generate induced MSCs (iMSCs), which may be useful for regenerative therapies.
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 3326. doi:1538-7445.AM2012-3326