bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4256: Use of short tandem repeat profiling for authentication of human cell lines
Authors: Reid Thomas M. , Wilson Keen , Kraemer Christopher , Baird Michael 
Published: August 2012

Abstract
Human cell lines are widely used in biological and biomedical research, and accurate data interpretation depends on unambiguous identity of a cell line relative to its original source. Contamination and misidentification of human cell cultures has been identified as a long-standing issue within the scientific community, and steps are now being taken to address the problem. Short-tandem-repeat (STR) DNA profiling provides an accurate, reliable, and standardized method for authentication of human identity. Currently, the ATCC is considering proposed standards for cell line authentication utilizing human STR typing, and many journals (including all AACR published journals) have adopted the NIH-recommended policy to mandate cell line authentication as a requirement for publication. Here we demonstrate the validation and use of two different STR systems to authenticate human cell lines as well as detect contamination by mouse feeder cells and other human cell lines. Threshold levels for detection of complete profiles as well as contamination are compared between the systems, and typical typing issues specific to cell lines are discussed.
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 4256. doi:1538-7445.AM2012-4256
Abstract 426: Sox2 expression in breast cancer stem cells: Possible mechanisms
Authors: Esnaola Olatz Leis , Eguiara Arrate , Jesus Alberdi Maria , Kepa Elorriaga , Rezola Ricardo , Martin Angel G. 
Published: August 2012

Abstract
The cancer stem cell (CSC) model does not imply that tumours are generated from transformed tissue stem cells. The target of transformation could be a tissue stem cell, a progenitor cell, or a differentiated cell that acquires self-renewal ability. The observation that induced pluripotency reprogramming and cancer are related has lead to the speculation that CSCs may arise through a reprogramming-like mechanism. Expression of pluripotency genes (Oct4, Nanog and Sox2) was tested in breast tumours by immunohistochemistry and it was found that Sox2 is expressed in early stage breast tumors. Mammosphere formation in culture was used to reveal stem cell properties, where expression of Sox2, but not Oct4 or Nanog, was induced. Over-expression of Sox2 increased mammosphere formation, effect dependent on continuous Sox2 expression; furthermore, Sox2 knockdown prevented mammosphere formation and delayed tumour formation in xenograft tumour initiation models. These findings suggest that reactivation of Sox2 represents an early step in breast tumour initiation, explaining tumour heterogeneity by placing the tumour-initiating event in any cell along the axis of mammary differentiation. Now we present evidence on what breast cancer specific mechanisms may control Sox2 expression activation.
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 426. doi:1538-7445.AM2012-426
Abstract 4263: Dynamic phenotype switching regulated by epigenetic marks along melanoma genesis
Authors: Morais Alice S. , Molognoni Fernanda , Jasiulionis Miriam Galvonas 
Published: August 2012

Abstract
Melanoma is the most aggressive form of skin cancer and its incidence is increasing worldwide. Although melanoma is highly curable when early diagnosed, the metastatic melanoma detains the worst prognosis and remains one of the most resistant cancers to treatment. In this light, continuous efforts are made to better understand the pathogenesis of this disease. It has been strongly suggested that aberrant epigenetic events, occurred in specific steps, are important to the establishment and progression of tumor genesis as the occurrence of point genetic mutations. Based on a murine model of malignant transformation, the aim of our group has been identify and understand the effective participation of epigenetic mechanisms in the generation of a transformed phenotype. The study model of melanoma genesis was established in vitro by submitting non-tumorigenic melanocytes (murine melan-a cell line) to sequential cycles of anchorage blockade, proposing that microenvironment interference may lead to intracellular alterations with important gene expression impact. Our cell lines represent distinct phases of melanoma genesis (melan-a melanocytes, 4C pre-malignant melanocytes, 4C11- non-metastatic and 4C11+ metastatic melanoma) and, on an overview, it was observed clearly morphological differences, mainly in intermediate stages of progression, suggesting a transient change from epithelial to mesenchymal phenotype (from melan-a to 4C and 4C11- cell lines) along malignant transformation. This transitory modification was accompanied by a switch in the EMT markers expression, like Snail1 and your targets, β-catenin, E-cadherin and N-cadherin. In parallel, Chd1 (chromatin remodeling factor), Nanog (pluripotency factor) and Jarid1b (histone demethylase) are highly expressed in these intermediate phases (4C and 4C11-), characterizing an open chromatin configuration. Interestingly, expression of most of these genes appear to be controlled by epigenetic mechanisms, both DNA methylation and chromatin modification. We hypothesized that during melanocyte malignant progression associated with sustained stress, an opened chromatin occurrence would facilitate cells to undergo changes in gene expression which allow cell adaptation to new environment and all these alterations are orchestrated by epigenetic events. Supported by FAPESP.
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 4263. doi:1538-7445.AM2012-4263
Abstract 4265: Human prostate mesenchymal stem cells in prostate cancer: Mechanisms of TGF-β1 promoted myofibroblast/CAF differentiation
Authors: Kim Woosook , Barron David , Ressler Steven , Rowley David 
Published: August 2012

Abstract
Reactive stroma in prostate cancer is typified by the co-evolution of myofibroblasts and carcinoma-associated fibroblasts (CAFs), yet the origin and biology of myofibroblast stem/progenitor cells are not known. TGF-β1 is overexpressed in prostate cancer cells and affects myofibroblast differentiation, a key step in formation of a tumor-promoting reactive stroma. However, specific mechanisms of how TGF-β1 regulates recruitment, activation and differentiation of myofibroblast stem/progenitor cells are poorly understood. Human prostate mesenchymal stem cell (hpMSC) lines were initiated from organ cultures of normal human prostate gland and characterized by a novel 3D co-culture model. Differentially engineered LNCaP cells to overexpress active TGF-β1 were cultured as organoids on CM-inserts and hpMSCs were co-cultured with engineered LNCaP cells either in direct contact or on laminin-coated coverslips in the bottom chamber. 3D organoids grown in serum-free media for 72 hours were analyzed by IHC, ELISA and qRT-PCR. In addition, RNAi gene knockdown experiments were performed in hpMSC to identify key transcription factors involved in myofibroblast/CAF differentiation. hpMSCs were CD44+/CD90+ and showed characteristics and gene expression similar to human mesenchymal stem cells. In the 3D model, LNCaP cells and hpMSCs self-organized into a free-floating organoid. The organoids exhibited a periphery of LNCaP cells and a core of reactive stroma-appearing cells. Significantly, LNCaP cells induced myofibroblast differentiation of hpMSC and this was promoted by LNCaP expression of active TGF-β1. Gene expression analysis identified the RUNX1 and ID1 transcription factors as upregulated in hpMSC by TGF-β1. Knockdown of RUNX1 and ID1 are being used to assess the role and mechanisms of how these transcription factors regulate myofibroblast differentiation. Together, these data suggest that the genesis and co-evolution of reactive stroma in prostate cancer results from activation and differentiation of CD44+/CD90+ endogenous hpMSC via upregulation of specific transcription factors by TGF-β1. Defining key mechanisms that recruit, activate and differentiate the origin of myofibroblasts/CAFs could provide clues to develop new strategies to target the microenvironment niche.
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 4265. doi:1538-7445.AM2012-4265
Abstract 4266: Identification of gene expression profiles associated with different types of breast adipose and their relationship to tumorigenesis
Authors: Field Lori A. , Deyarmin Brenda , van Laar Ryan , Shriver Craig , Ellsworth Rachel E. 
Published: August 2012

Abstract
Background: Research over the past decade has shown the importance of the stroma in tumorigenesis, however, having long been thought to function only as an inert energy storage depot, the role of adipose tissue in tumorigenesis has been largely ignored. Improved understanding of the role of adipose in tumorigenesis is crucial given the increasing rates of obesity and the use of autologous fat transfer in breast reconstruction. Methods: Adipose, adjacent to and distant from invasive breast tumors, was laser microdissected from 20 post-menopausal women, and from 20 post-menopausal women with non-malignant breast disease. Gene expression data were generated using U133 2.0 microarrays. After quality control and visualization steps, the data were analyzed to identify significant patterns of differential expression between adipose classes, at the individual gene and molecular pathway level. Results: Pathway analysis revealed that immune response differs between non-malignant, distant and tumor adjacent adipose; this response is seen as a gradient with the largest response closest to the tumor. Gene expression differed significantly in adipose from invasive compared to non-malignant breasts with FCGR2A, FOLR2, LGMN, MARCO and NLRP3 expressed at significantly higher levels and HLA-DQB1 and HLA-DQA1 at significantly lower levels in adipose from invasive breasts. Within the invasive breasts, MMP9, PLA2G7, RRM2 and SPP1 were expressed at &gt;3-fold higher levels in adjacent compared to distant adipose. Conclusions: Gene expression levels differ in breast adipose, depending on presence of or proximity to tumor cells. Adipose adjacent to the tumor demonstrated the largest immune response; this response may reflect a reaction to surgical insult from the original biopsy; however, response to surgical injury has been associated with increased ability to metastasize. In addition, within breasts with invasive breast cancer, genes involved in cellular proliferation, degradation of the extracellular matrix and angiogenesis were expressed at higher levels in adjacent compared to distant adipose. Together, these data suggest that adipose is not an inert component of the breast microenvironment but plays an active role in tumorigenesis.
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 4266. doi:1538-7445.AM2012-4266
Abstract 4273: C-kit expression promotes prostate cancer cell migration and invasion through Brca2 downregulation
Authors: Zhe Xiaoning , Azevedo Marcelo Quintanihla , Cher Michael L. , Kim Hyeong-Reh C. , Bonfil R. Daniel 
Published: August 2012

Abstract
In many human cancers, expression or activity of the receptor tyrosine kinase (RTK) c-kit is often associated with increased malignancy. In prostate cancer (PCa), we reported an association between expression of c-kit and clinical progression, with the highest expression seen in castrate-resistant metastatic bone specimens. We also found de novo c-kit expression in experimental intraosseous tumors formed by otherwise c-kit negative PCa cells. However, the biological consequences of c-kit expression and activation in PCa cells remain unknown. Here, we show that c-kit overexpression in PC3 and C4-2B PCa cells significantly enhances their migratory and invasive ability in vitro. Consistent with this result, we found that expression and activation of c-kit in PCa cells decreased the expression of E-cadherin and increased the expression of vimentin, suggesting an epithelial mesenchymal transition-like phenomenon. Furthermore, activation by c-kit ligand in PC3 and C4-2B cells transfected with c-kit lead to phosphorylation of Akt. This contributes to the enhanced invasiveness of c-kit-expressing PCa cells, as demonstrated by an inhibitory effect by PI3K/Akt inhibitors LY294002 and Wortmannin. In an attempt to identify genes differentially expressed in c-kit-expressing and -non-expressing PCa cells, we used a microarray containing 84 genes, representing growth factor receptors, transcription factors, cell cycle, and DNA repair genes. The analysis of the array demonstrated a consistent down-regulation of Brca2 in both c-kit-expressing PC3 and C4-2B cells, further validated by qRT-PCR and immunoblotting. Re-introduction of Brca2 into c-kit-expressing PCa cells suppressed c-kit-enhanced invasiveness to levels similar to those shown by c-kit-non-expressing cells, and led to reduced phosphorylation of Akt. Our results provide evidence that c-kit expression in PCa cells, an event which is induced upon interaction with the bone microenvironment, promotes tumor migration and invasion through an Akt-dependent mechanism regulated by Brca2 down-regulation. The identification of this novel signaling axis and its importance in PCa migration and invasion reveal potential therapeutic targets for regulating the expansion of PCa metastatic lesions in bone.
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 4273. doi:1538-7445.AM2012-4273
Abstract 4275: PLA2 activities in human epithelial ovarian cancer ascites
Authors: Xu Yan , Cai Qingchun , Zhao Zhenwen , Antalis Caryl J. , Yan Libo , Hamed Ali H. , Stehman Frederick B. , Schilder Jeanne M. , Priore Giuseppe Del 
Published: August 2012

Abstract
Ascites production is characteristic of late stage human epithelial ovarian cancer (EOC) and it correlates with tumor spread clinically. However, the role of this microenvironment in cancer cells and the mechanisms by which ascites promotes tumor development are not well understood. In recent years, phospholipase A2 (PLA2) enzymes have been identified as cancer therapeutic targets focusing mainly on secreted PLA2s (sPLA2s). In this work, we optimized and validated the quantitative analytic methods to measure PLA2 activities in human EOC ascites and tissues. We found, for the first time, that both cytosolic PLA2 (cPLA2) and calcium-independent PLA2 (iPLA2), but not sPLA2 and autotaxin (ATX), activities (not their expression levels) in human EOC tissues and ascites (the tumor microenvironment) were significantly elevated. In addition, while cell-free (S1) and vesicle-free (S4) human EOC ascites had potent promoting activities in proliferation, migration, and invasion of human EOC cells, the PLA2 activities were involved and account for a significant portion of these activities. Moreover, in vivo studies demonstrated that methyl arachidonyl fluorophosphonate (MAFP, a dual inhibitor of cPLA2 and iPLA2), was highly effective in inhibiting EOC tumorigenesis/metastasis in xenograft models, supporting PLA2 activities as new targets for EOC. LPA, an enzymatic product of ATX and PLA2 enzymes, mediates a significant part of the cellular effects of ascites as evidenced by 1) LPA is produced in cell-free ascites; 2) LPA production and their sensitivities to inhibitors correlate well with the cellular effects in ascites; and 3) the cellular activities were sensitive to LPAR inhibitor and siRNA against LPARs. While cPLA2 and iPLA2 are cytosolic enzymes, our findings indicate that they are associated with cell- and microvesicle-free ascites. In summary, our work implies the potential marker and therapeutic values of PLA2 activity in ovarian 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 4275. doi:1538-7445.AM2012-4275
Abstract 4281: Adaptive post-translational regulation of the thiamine transporter THTR1 in breast cancer cells during hypoxic stress
Authors: Zastre Jason , Sweet Becky , Govindarajan Rajgopal 
Published: August 2012

Abstract
Purpose: The cellular metabolic re-programming within hypoxic tumor microenvironments involves adaptations that alter the cellular dependence for glycolysis and the pentose phosphate pathway. However, enzymatic activity within these pathways requires an adequate supply and cellular pool of enzyme cofactors. In particular, thiamine is an essential cofactor within key metabolic pathways that are responsible for energy production and anabolic precursor synthesis. Previously, we have reported on the hypoxia-mediated transcriptional regulation of the thiamine transporters within breast cancer cells. Therefore, the purpose of this study was to further our understanding of hypoxia-mediated regulation of thiamine transporters, by examining the intracellular localization and plasma membrane expression of THTR1 during hypoxic stress. Methods: Immunofluorescence was employed to visualize the intracellular localization of THTR1 (SLC19A2) within the non-transformed breast epithelial cell line, MCF10A and breast cancer cells, BT20 and BT474 after 1, 3 and 7 days of 1% O2 exposure. Cell surface biotinlyation assays were utilized to assess changes in the plasma membrane expression of THTR1 after hypoxia exposure. In addition, total THTR1 expression was determined using Western blotting of whole cell lysates and qRT-PCR. Results: The cellular distribution of THTR1 was predominantly localized in the plasma membrane for the non-transformed MCF10A cells, but was predominantly intracellular in the breast cancer cells, BT20 and BT474. No change in total mRNA or cellular THTR1 protein expression was found after hypoxic exposure. However, THTR1 exhibited a time dependent increase in plasma membrane localization after 1% O2 exposure. This observation was confirmed with an increase in THTR1 plasma membrane expression using cell surface biotinylation pull down assays. Conclusions: We conclude that hypoxic stress induces the intracellular trafficking of the thiamine transporter THTR1 to the plasma membrane. These results suggest that post-translational modification of THTR1 during hypoxic exposure may be an essential adaptive cellular response to increase intracellular supply of thiamine to support glycolytic metabolism.
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 4281. doi:1538-7445.AM2012-4281
Abstract 4282: Analysis of epithelial and stromal cell networks in prostate cancer by gene expression profiles
Authors: Ashida Shingo , Kawada Chiaki , Inoue Keiji , Shuin Taro 
Published: August 2012

Abstract
Recent studies have highlighted tumor microenvironment as important in carcinogenesis and progression. Especially in prostate cancer (PC), it has been suggested that tumor microenvironment plays an important role in progression, acquisition of androgen independence, and distant metastasis. However, little is known about the genetic basis of human PC microenvironment. The prognosis of PC is related to invasion and metastasis. Epithelial-to-mesenchymal transition (EMT) has gained considerable attention as a conceptual paradigm to explain invasive and metastatic behavior during cancer progression. EMT is a normal physiologic process by which cells of epithelial origin convert into cells with mesenchymal characteristics. It has been proposed that EMT is induced by cancer cells during metastatic dissemination from a primary organ to secondary sites, but how EMT occurs during PC invasion and metastasis remains uncertain. To clarify the mechanism of PC progression and metastasis, we investigated cross-interaction of PC epithelial cells and stromal cells by analyzing genome-wide gene expression profiles. We hypothesized that PC cells could affect on stromal cells to change their characteristics into so-called cancer-associated fibroblasts (CAFs), which might contribute to cancer invasion and metastasis. Likewise, CAFs could affect on surrounding normal epithelial cells to change their characteristics into PC cells. We first analyzed three of genome-wide gene expression profiles; normal human prostate stromal cells (PrSC) co-cultured with human prostate cancer cells (LNCaP), LNCaP co-cultured with PrSC, and human prostate epithelial cells (PrEC) co-cultured with cancer-associated PrSC which is PrSC previously co-cultured with LNCaP. Before microarray analysis, we confirmed the overexpression of alpha-smooth muscle actin in PrSC co-cultured with LNCaP, which means the transition from normal stromal cells to CAFs. We then analyzed networks among PC cells, stromal cells, and surrounding normal epithelial cells by analyzing gene expression profiles and validated our data using real-time PCR, immunohistochemistry, and bioinformatics. These data provide clues to the molecular mechanisms underlying PC invasion and metastasis, and suggest candidate genes the products of which might serve as molecular targets for the treatment of PC.
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 4282. doi:1538-7445.AM2012-4282