bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3049: Overexpression of miR-17 family miRNAs in prostate cancer biopsies: Evidence for a stem-cell-like miRNA profile in high grade/high stage tumors
Authors: Gaston Sandra M. , Guerra Andrew L. , Nadadur Rangarajan C. , Meadows Julie , Kearney Michael C. , Sorbara Lynn , Srivastava Sudhir , Meltzer Stephen J. , Pinsky Paul , Kearney Gary P. 
Published: February 2011

Abstract
Complex changes in microRNA (miRNA) expression patterns have been observed in many different types of cancer, including prostate cancer. Although the consequences of deregulation of miRNA expression in human cancer are still largely unknown, the use of miRNAs for cancer classification and prognosis has emerged as an important new area of biomarker research. We compared the tissue expression of miRNAs in prostate biopsies from patients with high grade/high stage tumors with biopsies from patients with no evidence of malignancy. Snap frozen prostate biopsy samples were prospectively collected using tissue printing techniques and total RNA was isolated for miRNA analysis using Agilent miRNA microarrays (Version 1, containing 470 human tissue miRNAs). The prostate cancer cases included in the microarray profiling study had at least 5 of 12 cores positive for cancer and significant areas of Gleason 7 or more. The no-cancer controls showed no prostate cancer, HGPIN or suspicious histology in any of the 12 biopsy cores. In our analysis of miRNA expression patterns, “significant change in expression” was defined as at least 2 fold change and F test (p&lt;0.05). Three different internal reference normalization methods were used in the data analysis; here we classify miRNAs as “overexpressed” or “underexpressed” only when all three methods agree. In high grade/high stage biopsy cancer, 64 of the 470 human tissue miRNAs on the microarray (13.6%) were found to be significantly overexpressed when compared with miRNA expression in biopsies from no-cancer controls. This set of overexpressed miRNAs includes 4 of the 5 miR-200 family miRNAs that have been implicated in the regulation of the epithelial-to-mesenchymal transition (EMT). The cancer-overexpressed miRs also include 6 of the 8 miR-17 family genes; this miRNA family has been implicated in the control of stem-cell differentiation during embryogenesis. Interestingly, the 6 miR-17 family miRs that are overexpressed in high grade/high stage prostate cancer include 2 from each of the 3 polycistronic clusters that encode this highly conserved miR family, an expression pattern similar to the coordinated expression observed in early embryonic development. This robust stem-cell-like miRNA profile offers an interesting new candidate biomarker that may be particularly sensitive to an aggressive prostate cancer phenotype.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3049.
Abstract 3050: MicroRNA expression profiling of human Barrett's carcinogenesis
Authors: Fassan Matteo , Volinia Stefano , Palatini Jeff , Pizzi Marco , Baffa Raffaele , Clemente Roberto , Rizzetto Christian , Croce Carlo M. , Zaninotto Giovanni , Ancona Ermanno , Rugge Massimo 
Published: February 2011

Abstract
Barrett's Mucosa (BM) is a metaplastic replacement of the native esophageal (squamous) epithelium (ESq) by columnar-intestinalized mucosa. BM is the main risk factor for Barrett's adenocarcinoma (BAc). MicroRNAs (miRNAs) are a class of small non-coding RNAs that control gene expression by targeting mRNAs; miRNAs deregulation has been associated with Barrett's oncogenesis. To explore the hypothesis that a specific miRNAs signature is associated with BM (and its associated cancer risk) a miRNA microarray analysis (OSU-CCC version 4.0; Ohio State University) compared ESq versus all the spectrum of the phenotypic lesions of the Barrett's carcinogenesis. Specimens were collected at Department of Pathology of Padova University from 14 patients undergone esophagectomy for BAc or High-grade non-invasive neoplasia (HG-NiN). Tissues samples for the microarray study included: ESq= 14 samples; BM= 14; Low-grade non-invasive neoplasia (LG-NiN)= 7; HG-NiN= 5; BAc= 11. The miRNA-profiling study consistently disclosed increased expression of 6 miRNAs (hsa-miR-215, hsa-miR-560, hsa-miR-615-3p, hsa-miR-192, hsa-miR-326, hsa-miR-147), and decreased expression of 7 others (hsa-miR-100, hsa-miR-23a, hsa-miR-605, hsa-miR-99a, hsa-miR-205, hsa-let-7c, hsa-miR-203). Microarray results were further validated by qRT-PCR, using a different series of 10 consecutive BAc cases (Esq= 10 samples; BM= 6 samples; BAc= 10 samples). Interestingly, some of the miRNAs found deregulated in this series are already known as suitable markers of cancer progression in other models of epithelial oncogenesis (i.e.: hsa-let-7c, hsa-miR-192, hsa-miR-203, hsa-miR-205, hsa-miR-215). The results achieved by the miRNA profiling study were further validated by the immunohistochemical (IHC) analysis on a well-defined miRNA gene target (HMGA2 [hsa-let-7c]). The IHC expression of the protein product was consistent with the corresponding miRNA's deregulation. In fact, HMGA2 IHC expression was up-regulated in HG-NiN and BAc. The achieved results confirm that specific miRNAs are involved in BM carcinogenesis and that they may represent a novel diagnostic/prognostic tool in the characterization of BAc gene targets.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3050.
Abstract 3057: The serine protease inhibitor, elafin, exhibits novel tumor suppressor functions in the context of breast cancer
Authors: Caruso Joseph A. , Hunt Kelly K. , Keyomarsi Khandan 
Published: February 2011

Abstract
Serine proteases and their inhibitors have an established role in cancer progression, invasion, and metastasis. Elafin is an endogenously expressed inhibitor of human neutrophil elastase. In mammary epithelial cells elafin is highly expressed; however in breast cancer it is transcripitionally downregulated. We hypothesize that the loss of elafin is critical to the development of the malignant phenotype in breast cancer. In human mammary epithelial cells (HMECs) and breast cancer cell lines, the regulation and activity of elafin is dependent on the critical tumor suppressor retinoblastoma (Rb). Elafin is dramatically upregulated in quiescent HMECs following growth factor deprivation. This upregulation is characterized by an Rb dependent increase in elafin stability and cytoplasmic accumulation. Knockdown of elafin by shRNA in HMECs causes them to be refractory to growth factor deprivation induced quiescence; suggesting that elafin upregulation is a key regulator of cellular quiescence. Overexpression of elafin by adenovirus in both HMECs and breast cancer cell lines lacking Rb causes caspase-3 dependent, apoptotic cell death. Additionally, adenoviral elafin causes Rb-dependent cell cycle arrest in tumor cells with functional Rb. The Rb pathway is lost or inactivated in the majority of cancers. The design of a therapeutic strategy capable of specifically targeting tumor cells with Rb inactivation may prove to have significant utility in the treatment of cancer. The data summarized here demonstrates that overexpression of elafin is cytostatic in Rb-positive breast cancer cells and cytotoxic in Rb-negative breast cancer cells expressing caspase 3; suggesting that elafin could be engineered into a therapeutic modality in the treatment of breast cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3057.
Abstract 3062: Identification and functional analysis of NOL7 tumor suppressor gene nuclear and nucleolar localization signals
Authors: Lingen Mark W. , Zhou Guolin , Doci Colleen 
Published: February 2011

Abstract
NOL7 is candidate tumor suppressor found within the nucleus and nucleolus. However, the mechanisms regulating its subcellular localization have not been elucidated. An in vitro import assay demonstrated that NOL7 requires cytosolic machinery for active nuclear transport. Using NOL7 deletion constructs and cytoplasmic pyruvate kinase (PK) fusion proteins, three nuclear localization signals (NLSs) were identified. Site-directed mutagenesis of PK fusions and full-length NOL7 defined the functional regions within each NLS. Further characterization revealed that NLS2 and NLS3 were critical for both the rate and efficiency of nuclear targeting. In addition, four basic clusters within NLS2 and NLS3 were independently capable of nucleolar targeting. The dynamics of nucleolar localization were investigated by FRAP and specific nucleic acid depletion. NOL7 is relatively immobile within the nucleolus, but shuttles rapidly between the nucleolus and nucleoplasm. In addition, targeting to the nucleolar compartment was dependent on the presence of RNA, as depletion of total RNA or rRNA resulted in a nucleoplasmic shift of NOL7. Take together, these results identify the minimal sequences required for the regulation of NOL7 subcellular localization, characterizes the contribution of each sequence to NOL7 nuclear and nucleolar dynamics and suggests a functional role for NOL7 in both compartments.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3062.
Abstract 3063: Novel putative tumor suppressor LZAP binds and regulates Wip1
Authors: An Hanbing , Lu Xinyuan , Yarbrough Wendell G. 
Published: February 2011

Abstract
Our group has shown that LZAP is lost in ∼30% of head and neck squamous cell carcinomas and that inhibition of LZAP expression is associated cellular transformation and xenograft tumor growth. LZAP increases invasion and inhibits cellular proliferation and clonogenic growth mediated correlating with inhibition of NF-kB and activation of p53. Other researchers have found that LZAP promotes cell death in response to cytotoxic therapy at least partially dependent on activation of Cdk1 through Chk1/2 inhibition. We have also found that LZAP binds and inhibits p38 and this inhibition does not depend on upstream regulators MKK3 and MKK6. The activity of MAPKs reflects a balance between the upstream activating kinases and inactivating protein phosphatases. Since LZAP inhibition of p38 is not through upstream kinases, we determined if phosphatase activity targeting p38 was regulated by LZAP.
Wip1 is a major phosphatase regulating p38 phosphorylation and activation. Wip1 is a member of the PP2C family of serine/threonine phosphatases and was identified in a screen to detect p53 responsive genes. We noted that LZAP and Wip1 had many binding partners/targets including RelA, p38, and Chk1/Chk2 in common. Mechanisms of LZAP inhibition of RelA, p38 and Chk1/Chk2 are not well described, but expression of LZAP is associated with decreased phosphorylation of all three target proteins. To determine if LZAP could regulate Wip1 activity, we first evaluated LZAP and Wip1 interaction through immuno- or affinity precipitation. LZAP and Wip1 co-precipitated after expression in mammalian cells and bacterially expressed LZAP pulled down Wip1 generated through in vitro transcription and translation. After transient expression in mammalian cells, Wip1 and LZAP co-localized by indirect immunofluorescence staining. When expressed singly, Flag-tagged Wip1 localized to the nucleus and LZAP was excluded from the nucleolus, but localized to both cytoplasm and nucleus. When Wip1 and LZAP were co-expressed LZAP localization was altered so that nucleolar LZAP was observed. These findings indicate that co-expression of LZAP and Wip1 results in alternation of LZAP subcellular localization with resultant co-localization of these two proteins to nucleus. Functionally, co-expression of LZAP with Wip1 increased Wip1 content within p38 immunocomplexes which was associated with decreased p38 phosphorylation.
Together, our findings indicate LZAP binds and co-localizes with Wip1 and increases activity of Wip1 towards p38. Wip1 may play a common mediator for LZAP activities.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3063.
Abstract 3066: YPEL3, a novel inducer of senescence, is inhibited by estrogen in ER+ mammary tumor cells
Authors: Tuttle Rebecca M. , Miller Kelly L. , Termulhen Paula M. , Berberich Steven J. 
Published: February 2011

Abstract
It was recently discovered by this laboratory that Yippie-like 3 (YPEL3) is a novel p53 regulated gene that when induced triggers premature senescence in primary and tumor cell lines. Initial screens of human tumor samples uncovered significant decreases in YPEL3 gene expression in ER- verses ER+ breast cancers leading us to believe that estrogen may have a regulatory effect on YPEL3 expression. Using ER+ MCF7 cells, we determined that estrogen is a significant inhibitor of YPEL3 gene expression in cells grown in media with 10% serum verses cells grown in media with charcoal stripped serum (CSS) from which estrogen has been removed. The increase in YPEL3 mRNA expression demonstrated in CSS can be blocked by the addition of estrogen, and as expected tamoxifen competes with this estrogen repressive effect. The effects of estrogen on YPEL3 expression are not observed in MCF10A mammary epithelial cells which are ER negative demonstrating the requirement for the estrogen receptor in the observed effect. Given our observation that YPEL3 induction triggers senescence in breast cancer cells, we employed beta-galactosidase activity as a measurement of senescence and observed an 8-fold increase in senescence in MCF7 cells grown in CSS media. The increase is reduced to less than 3.5-fold with the addition of estrogen to the CSS media, and the increase in cellular senescence is blocked in MCF7 cells stably selected for the loss of YPEL3 (MCF7-shYPEL3) suggesting the need for the absence of estrogen and the presence of YPEL3 in senescence induction. We are currently testing the effects of estrogen on MCF7 cells where p53 has been knocked down using RNAi approaches and are planning to test the effects of RNAi targeting ER-alpha. We believe estrogen is inhibiting YPEL3 expression through its association with the estrogen receptor and their association with p53 thereby inhibiting p53 transactivation of YPEL3. These findings suggest that modulation of YPEL3 by estrogen and its growth suppressive effects may provide a target for future breast cancer therapies.
Note: First two authors contributed equally.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3066.
Abstract 3069: Importance of zinc-binding domain of matrix metalloproteinase 19 (MMP19) in tumor suppression and anti-angiogenesis of nasopharyngeal carcinoma
Authors: Chi Chan King , Ko Josephine M.Y. , Lung Hong Lok , Sedlacek Radislav , Lung Maria Li 
Published: February 2011

Abstract
Background and aims: Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases consisting of more than 20 enzymes. MMP19 shares the typical MMP structural domains. It degrades various ECM components including collagen type IV, nidogen-1, fibronectin, tenascin-C isoform, aggrecan, and laminin-5-gamma-2-chain. In our previous studies, MMP19 was identified as a candidate tumor suppressor gene (TSG) in nasopharyngeal carcinoma (NPC). It was down-regulated in NPC cell lines and NPC biopsies, when compared to an immortalized normal nasopharyngeal epithelial cell line and corresponding non-tumor nasopharyngeal tissues. Transfection of wild-type (WT) MMP19 into the HONE1-2 NPC cell line significantly suppressed tumor formation in the nude mouse tumorigenicity assay and decreased the tube-forming ability of both human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1). In this study, we aimed to validate the importance of the zinc-binding domain of MMP19 in NPC. Methods: We transfected the mutant MMP19, which contained a mutation at the zinc-binding domain, into HONE1-2 to study the effect of mutant MMP19 in tumor suppression and anti-angiogenesis compared to WT MMP19. Results: Mutant MMP19 was not tumor-suppressive, as is the WT MMP19, and aggressive tumors appeared with the same growth kinetics as the vector-alone control. The anti-angiogenesis effect was also not observed in endothelial cells incubated with conditioned medium from mutant MMP19 clones compared to WT MMP19 clones. Conclusions: Previous studies showed mutation of the zinc-binding domain in MMP19 leads to loss of MMP19 function in cell proliferation, migration, and protein degradation in a human keratinocyte cell model. This study demonstrates loss of tumor suppressive and anti-angiogenesis function of mutant MMP19 in NPC and indicates the importance of the zinc-binding motif for WT MMP19 in suppression of tumor formation and in inhibition of tube-forming ability of endothelial cells in NPC.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3069.
Abstract 307: EphB4 promotes or suppresses Ras/ERK pathway depending on cellular contexts: Implications for EphB4 as a cancer target
Authors: Xiao Zhan , Carrasco Rosa A. , Kinneer Krista , Sabol Darrin , Coats Steve , Jallal Bahija , Tice David 
Published: February 2011

Abstract
EphB4 is a member of the Eph RTK family. Its classical role has been shown to act in neuronal guidance and mediate venal/arterial separation. In contrast to these more established roles, EphB4's function in cancer is controversial since both tumor promoting as well as tumor suppressing functions have been ascribed to it. Here we provide a vivid illustration of this intriguing dichotomy by showing that EphB4 activation resulted in inhibition of the Ras/ERK pathway in endothelial cells but induction of the same pathway in MCF-7 breast cancer cells. This was true if EphB4 was stimulated with ephrinB2, its natural ligand, or an EphB4-specific fully human monoclonal antibody. The reason for these dramatic differences is due to potential functional coupling of EphB4 to different downstream effectors. In HUVEC cells, the RNAi knockdown of p120 RasGAP, a Ras inhibitor, abrogated the inhibitory effect on Ras/ERK pathway. However, in MCF7 cells, RNAi knockdown of PP2A, which has been shown to activate the Ras/ERK pathway through removal of the inhibitory phosphorylation on c-Raf, negated EphB4-driven activation of the Ras/ERK pathway. This represents the first time a functional coupling between Eph receptor and PP2A has been established, and this interaction appears to confer activation but not suppression of an oncogenic pathway. Our study also demonstrates the caveats and potential challenges of targeting EphB4 for cancer therapy due to the presumed conflicting effects on cancer cell and endothelial cell compartments.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 307.
Abstract 3074: Neutrophil gelatinase-associated lipocalin inhibits proliferation of human esophageal adenocarcinoma cells both in vitro and in vivo
Authors: Tong Zhimin , Koolwal Pooja , DeMars Catherine J. , Buttar Navtej S. , Guha Sushovan 
Published: February 2011

Abstract
Background: Neutrophil gelatinase-associated lipocalin (NGAL), a member of lipocalin family, is a 25-kDa secreted glycoprotein. NGAL modulates multiple biological functions including cell proliferation, invasion, and metastasis. NGAL is upregulated in multiple gastrointestinal cancers including colorectal, pancreatic, and esophageal adenocarcinoma (EAC). Our group recently showed that NGAL over-expression significantly blocked metastasis of pancreatic adenocarcinoma cells [Cancer Research 68(15): 6100-6108, 2008]. However, the biological role of NGAL in EAC is not known.
Aim: To examine the biological role of NGAL in human EAC cells both in vitro and in vivo.
Methods and Results: We used OE33 (well-differentiated) and Flo-1 (poorly differentiated) EAC cell lines as our model system. We observed that OE33 cells have significantly higher levels of NGAL expression than Flo-1 cells both by qPCR and western blot analyses. To further study the biological role of NGAL in EAC, we created stable NGAL over-expression clones in Flo-1 (Flo1-NGAL) and NGAL under-expression clones in OE33 (OE33-shNGAL) using lentiviral system. We observed that under-expression of NGAL in OE33 significantly increased cell proliferation as determined by clonogenic assays. In contrast, NGAL over-expression in Flo-1 significantly suppressed cell proliferation. We also observed that NGAL under-expression increased cyclin D1 expression, a known cell cycle regulator. This, in part, explained increased EAC cell proliferation secondary to NGAL under-expression. In addition, NGAL over-expression potently suppressed and conversely, NGAL under-expression significantly increased invasion of EAC clones through MatrigelTM in vitro. Furthermore, to confirm our in vitro findings, we observed that OE33-shNGAL clones strikingly increased tumor volume in a subcutaneouos heterotopic xenograft model in nude mice.
Conclusion: We demonstrated for the first time that NGAL under-expression increased cell proliferation and invasion in vitro, which could be in part secondary to regulation of cyclin D1 expression. NGAL under-expression also enhanced tumor growth in vivo. Thus, modulation of NGAL can be a novel therapeutic target in progression of EAC.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3074.