bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1242: A broad activity screen in support of a chemogenomic map for kinase signaling research and drug discovery
Authors: Gao Yinghong , Davies Stephen , Augustin Martin , Harvey Kevin J. 
Published: August 2012

Abstract
Since protein kinases play a fundamental role in cell signaling, especially as components of critical growth and differentiation pathways linked to cancer, kinase inhibitors have been commercialized as both therapeutics and research tools. This has been enabled to some degree by the chemical tractability of protein kinase ATP binding pockets, the automation of activity measurements, and the correlation of in vitro, cellular, and in vivo activities. Despite the development of a number of efficacious kinase inhibitors, the strategies for rational design of these compounds have been limited by target promiscuity. In an effort to better understand the nature of structure and activity across the kinome, especially as it relates to off-target effects, we screened a well-defined collection of kinase inhibitors using “gold standard” radiometric assays for inhibitory activity toward 234 kinases, which represent all branches of the kinome tree. We screened 158 small molecules initially identified in the literature as potent and specific inhibitors of kinases important from a therapeutic target as well as a signal transduction biology perspective. We performed hierarchical clustering of these benchmark kinase inhibitors based upon their kinome activity profiles and illustrate how they relate to chemical structure similarities; this provides new insights into inhibitor specificity and potential applications for probing new targets. Additionally, we developed a selectivity score for each kinase which we believe reflects general binding site accessibility to small molecules. Knowledge of such scoring indices can play an important role in therapeutic target selection and drug discovery strategies. Using this broad set of data, we provide a framework for assessing polypharmacology. Examples will be shown to demonstrate that we not only identify likely off-target inhibitor activities but also potential new uses for known small molecules. Moreover we demonstrate how activity data can be used to help explain the effects of small molecules observed in phenotypic screening.
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 1242. doi:1538-7445.AM2012-1242
Abstract 1245: Hepatitis B virus X protein induces Src homology 2-containing protein tyrosine phosphatase, SHP2, expression through the NF-kB pathway in HBV-infected hepatocellular carcinomas
Authors: Kim Hyun Mi , Kim Ga-hye , Yu Eunsil , Choi Jene 
Published: August 2012

Abstract
Hepatitis B virus X protein (HBx) has a crucial role in the development of hepatocellular carcinoma and activates many transcription factors, including AP-1, NF-kB and CREB. In the present study, we showed that increased SHP2 expression was observed in 16 (53%) of the 30 cases of HBV-positive hepatocellular carcinomas tissues. We investigated the relationship of SHP2 and HBx in stably HBx-overexpressing Huh7-X and transiently HBx expressing HEK293 cells. The level of SHP2 proteins was concomitantly increased with HBX in a dose-dependent manner in HEK293 cells. We identified a putative NF-kB binding site within 500 bp of the SHP2 promoter and also observed the decrease of SHP2 proteins after the treatment of the NF-kB/p65 inhibitor, parthenolide in HBx expressing cell. In addition, we confirmed that the complex of NF-kB and HBx proteins direct bound to the SHP2 promoter by chromatin immunoprecipitation (ChIP)-real time PCR assays. Taken together, we conclude that HBV infection induces SHP2 overexpression through the HBx-NF-kB pathway. Currently, we are investigating the effects of SHP2 overexpression in the proliferation of hepatocellular carcinoma 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 1245. doi:1538-7445.AM2012-1245
Abstract 1247: ARHI downregulates PI3K and Ras-MAP signaling and induces autophagy by promoting integrin and EGFR membrane protein degradation
Authors: Lu Zhen , Liao Warren S.L. , Yang Maojie , Gao Fengqin , Bast Robert C. 
Published: August 2012

Abstract
Autophagy selectively degrades cellular proteins, protein aggregates and organelles, providing energy in a nutrient-deprived environment. We have reported that autophagy can be induced by re-expression of ARHI (DIRAS3), a maternally imprinted Ras-related tumor suppressor gene that is down regulated in more than 60% of ovarian cancers. Whereas prolonged expression of ARHI in cultured cells induces autophagic cell death, ARHI expression in xenografts sustains dormant cancer cell survival for weeks. When ARHI levels were reduced in dormant cells, xenografts grew rapidly. Inhibition of ARHI-induced autophagy with chloroquine dramatically reduces regrowth of xenografts, suggesting that autophagy contributes to the survival of dormant cells. Re-expression of ARHI inhibits both PI3K and Ras-MAPK signaling. Growth factors (VEGF, IL-8, IGF) and ECM components (fibronectin) found in the cancer microenvironment can rescue ovarian cancer cells from ARHI-induced autophagic cell death in culture, associated with partial reversal of ARHI-mediated inhibition of PI3K signaling. Consequently, we have hypothesized that autophagy together with growth factors and ECM in the microenvironment are all required to maintain dormancy, where autophagy provides energy for xenografts and survival factors prevent autophagic death by partially reversing inhibition of PI3K-AKT signaling. The mechanism by which ARHI regulates PI3K and Ras-MAPK signaling has not been fully elucidated. Here, we show that re-expression of ARHI in SKOv3 ovarian cancer cells inhibits expression of integrin β1 and EGFR. ARHI expression enhanced EGF-mediated receptor endocytosis and colocalization of EGFR with Rab11 and Rab7, markers for multivesicular endosomes and lysosomes, respectively. Our results suggest that ARHI enhances membrane receptor turnover, in part, by facilitating the internalization and association of membrane receptors with vesicular trafficking proteins targeted for degradation in the lysosome. ARHI-mediated downregulation of membrane receptors reduces both PI3K-AKT and Ras-MAPK signaling, ultimately contributing to the inhibition of tumor growth.
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 1247. doi:1538-7445.AM2012-1247
Abstract 1254: FAK activity regulates αvβ5 integrin and osteopontin expression to control breast and ovarian cancer anchorage-independent growth
Authors: Tancioni Isabelle , Lawson Christine , Miller Nichol L.G. , Uryu Sean , Jean Christine , Chen Xiao L. , Lim Ssang-Taek , Ward Kristy , Schlaepfer David D. 
Published: August 2012

Abstract
Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that controls cell motility and survival downstream of integrin receptors. FAK expression and activity are elevated during breast and ovarian cancer progression and this is associated with a poor clinical prognosis. Small molecule FAK inhibitors (FAK-I) block breast and ovarian tumor progression through a novel mechanism that is associated with the apoptosis under anchorage-deprived conditions. Metastatic cells can grow as anchorage-independent spheroids and we hypothesize that a spheroid micro-environment remains dependent on integrin- and FAK-associated signaling for survival. Here we show that intraperitoneal (ip) injection of murine ID8 ovarian carcinoma cells into mice, recovery of ascites-associated tumor cells (ID8ip) and growth ex vivo, yielded tumor cells with elevated levels of phosphorylated FAK (Y397 and Y576 phosphorylation), αvβ5 integrin, and osteopontin (OPN) matrix protein expression compared to parental ID8 cells. Analyses of paraffin-embedded breast and ovarian tumor sections revealed significantly higher pY397 FAK (p&lt;0.05), β5 integrin (p&lt;0.0001), and OPN expression (p&lt;0.005) in stage II-III compared to stage I tumors. Treatment of ovarian ID8ip, human ovarian HEY carcinoma, or human MDA-MB-231 breast carcinoma cells with FAK-I inhibited cell growth, prevented FAK Y397 phosphorylation, reduced β5 integrin and OPN levels independent of effects on β1 or β3 integrins or changes in matrix proteins such as fibronectin. FAK-I impairs anchorage independent cell growth and orthotopic tumor growth but not growth of cells in plastic-adherent cell culture. Stable knockdown of either FAK or β5 integrin in HEY cells prevented anchorage-independent growth. Re-expression of wildtype or kinase-dead FAK within HEY cells verified the importance of FAK activity in promoting β5 integrin and OPN expression associated with enhanced tumor growth. Ongoing studies are focused on determining the molecular connections of FAK to the regulation of αvβ5 and OPN expression. Together, our studies support a novel role for a signaling loop involving OPN and αvβ5 integrin leading to FAK activation and reinforcement of OPN and β5 integrin expression within the spheroid and tumor micro-environment enhancing breast and ovarian cancer cell survival and tumor progression. Supported by Susan G. Komen for the Cure (KG111237) and NIH grant CA102310
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 1254. doi:1538-7445.AM2012-1254
Abstract 1257: Microarray analysis of BRONJ-like oral mucosal lesions in mice
Authors: Bae Susan , Williams Drake , Oh Ju Eun , Shi Songtao , Kang Mo , Shin Ki-Hyuk , Park No-Hee , Kim Reuben H. 
Published: August 2012

Abstract
Bisphosphonates (BPs) are potent anti-resorptive agents that are widely used to treat bone-associated diseases including metastatic bone cancer. Many reports have indicated that long-term use of BPs can develop BP-related osteonecrosis of the jaw (BRONJ) that occurs largely at the site of tooth extraction; however, the pathophysiology of BRONJ is still poorly understood. Clinically, BRONJ is defined as exposed necrotic bone with unhealed and open oral mucosa, implying that the lack of wound closure may be caused either directly by BPs or indirectly via biological signals (i.e., cytokines) from the BP-affected surrounding structures. We hypothesize that oral mucosal tissues lose their capacity to close the wound properly at the affected sites and that the genes involved in wound healing processes are differentially expressed between BRONJ lesions and the normal counterparts. To test our hypothesis, we utilized the established BRONJ mice model. Zometa was administered via tail vein 1 week prior to tooth extraction. After extracting the upper first molar, the extracted socket was allowed to heal, and the overlaying oral mucosal tissues were harvested after 2 weeks from control and BRONJ groups (n=3). The fresh tissue samples were subjected to RNA isolation, cDNA sysnthesis, and cDNA hybridization onto microarray chip (Affymetrix MG 430 2.0). The data analysis by functions using IPA software revealed that the genes involved in skeletal and muscular disorders, dermatological diseases, cellular movement, cellular assembly and organization, and inflammatory disease were significantly associated with BRONJ. In particular, we found that growth factors (i.e., Igf2, Fgf7, Pdf2bp2), collagens (i.e., Col1a1, Col1a2, Col2a1, Col2a2, Col5a1, Col5a2, Col11a1, and Col11a2), and non-collagenous proteins (i.e., DMP-1, ITBP, and SPP1) were significantly down-regulated in BRONJ groups. Taken together, our study suggests that the diminution of growth factors as well as collagenous and non-collagenous structural proteins may be associated with the development of BRONJ. This study was supported by NIDCR/NIH (R03DE021114 and K08DE017121).
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 1257. doi:1538-7445.AM2012-1257
Abstract 1270: Identification and functional analysis of Pim-1 kinase substrates in prostate cancer
Authors: Barkley Nicole , Li Xiang , Bieberich Charles 
Published: August 2012

Abstract
Protein kinases constitute one of the largest and most pleiotropic families of proteins. Mutations leading to activation of protein kinases and loss of function of phosphatases can have deleterious consequences that lead to disease. Pim-1, a serine/threonine kinase, is one of many kinases that may play a role in the etiology of malignant diseases including lymphoma and prostate cancer. Recent studies indicate Pim-1 is overexpressed in prostate cancer and is a highly plausible therapeutic target. Paradoxically, a decrease in Pim-1 expression in later stages of prostate cancer correlates with poor prognosis. Pim-1 has been known to be an oncogene for more than two decades, however, our understanding of how it functions is incomplete. A key underlying factor in this lack of knowledge is that, to date, fewer than fifteen Pim-1 substrates have been identified. The identification and validation of novel Pim-1 substrates could elucidate biomarkers of Pim-1 activity and provide insight into the molecular basis of Pim-1 function. A recent, innovative kinase-substrate profiling method termed the Reverse In-gel Kinase Assay (RIKA) reverses the role of kinase and substrate in a traditional in-gel kinase assay. The assay involves the partial immobilization of an active kinase in a denaturing polyacrylamide gel to determine which substrates from cell lysates can be phosphorylated by the kinase within the gel. The potential substrates are excised from the gel and identified using mass spectrometry. Using a Pim-1 RIKA approach, 40S Ribosomal Protein 7 (RPS7) was identified as a potential novel Pim-1 substrate. RPS7 was expressed in E.coli to conduct preliminary in vitro studies. These in vitro studies demonstrated that Pim-1 robustly phosphorylates RPS7. Further studies were performed to identify sites of phosphorylation on RPS7, and alanine-substitution mutants were generated that completely abolished phosphorylation of RPS7 by Pim-1. These data indicated that RPS7 may be a novel Pim-1 substrate. Studies to validate RPS7 as a direct target of Pim-1 in vivo, and to determine the functional consequences of Pim-1 phosphorylation of RPS7 are in progress.
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 1270. doi:1538-7445.AM2012-1270
Abstract 1271: Aberrant activation of Spleen Tyrosine Kinase in ovarian cancer identified through a global phosphorylation profiling of protein tyrosine kinases
Authors: Shoni Melina , Du Jinyan , Yang Junzheng , Ng Shu-Kay , Muto Michael George , Welch William , Crum Christopher , Berkowitz Ross , Golub Todd , Ng Shu-Wing 
Published: August 2012

Abstract
Aberrations in protein kinase activity and the consequent deregulation of downstream signaling pathways are considered a hallmark of oncogenesis. The purpose of this study was to investigate the phosphorylation state, the ultimate readout of dysregulated activity, of protein tyrosine kinases in ovarian tumors. An innovative Luminex multiplex platform was employed to systematically determine phosphorylation status of protein tyrosine kinases in a total of 69 protein lysates derived from normal human ovarian surface epithelial (HOSE) cells, ovarian tumors, and cancer cell lines. Significance Analysis of Microarrays (SAM) was performed to identify the signature of phosphorylated kinases with statistical significance. Profiling results were confirmed by using phosphor-specific kinase antibodies in immunoblot analysis of the profiled lysates and immunohistochemistry on formalin-fixed paraffin-embedded archived ovarian and fallopian tissues, as well as immunofluorescence of ovarian cancer cells growing as monolayer or 3D Matrigel cell cultures. A signature of twenty-two phosphorylated protein tyrosine kinases was identified by SAM analysis of the profiling data. Spleen Tyrosine Kinase (SYK), a novel tyrosine kinase included in the signature, was found significantly phosphorylated in lysates from ovarian cancer tissues compared to normal tissues by both Western blot analysis and immunohistochemistry on 94 clinical ovarian tissues (P=0.024). SYK phosphorylation was also identified in tubal intraepithelial carcinomas, the hypothesized precursors of high-grade serous ovarian carcinomas. However, SYK activation was not observed in ovarian cancer cell lines growing as attached monolayer cells, but was present when they grew as mouse xenografts or in 3D Matrigel in vitro cultures. We are the first to report that SYK, an extensively investigated tyrosine kinase in hematopoietic cells, is aberrantly phosphorylated and activated in ovarian cancer. SYK phosphorylation in ovarian cancer might be induced by extracellular stimuli that are exclusively present in the tumor microenvironment. These findings provide additional options in targeted therapy for 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 1271. doi:1538-7445.AM2012-1271
Abstract 1284: Estrogen receptor alpha transcriptionally regulates p53 in MCF7 human breast cancer cells
Authors: Berger Crystal , Qian Yingjuan , Chen Xinbin 
Published: August 2012

Abstract
The p53 tumor suppressor plays a critical role in maintaining genome stability. In response to genotoxic stress, p53 induces target genes for cell cycle arrest, apoptosis, and DNA repair. While p53 is the most commonly mutated gene in all cancers, it is only mutated in 20% of breast cancers. Interestingly, 70% of all breast cancer cases are estrogen receptor (ER) positive and are associated with wild type p53 expression. An ER-positive breast cancer diagnosis generally indicates good patient prognosis and treatment responsiveness with anti-estrogens such as tamoxifen. The relationship between ERα and p53 remains poorly defined. Here, we found that ERα positively regulates p53 expression. Specifically, knockdown of ERα decreases the amount of p53 along with its downstream target p21, a regulator of cell cycle arrest, in MCF7 breast cancer cells. Furthermore, we found that ERα transcriptionally regulates p53 expression via binding to the p53 promoter. Luciferase promoter reporter experiments showed that two estrogen receptor element half sites located at nt −1828 and −1611 from the p53 transcriptional start site responded to addition of ERα with increased reporter activity. Mutation of those two sites resulted in no reporter activity. To determine the biological significance of ERα regulation of p53, we found that knockdown of ERα desensitized cells to growth arrest and cell death upon treatment with DNA damage-inducing drugs in a p53-dependent manner. Conversely, cells overexpressing ERα were sensitized to growth arrest and cell death upon DNA damage-inducing drugs via the p53 pathway. Together, these results provide an insight into the reciprocal regulation of p53 and ERα at the transcript level and how their pathways interact in ER-positive breast cancers under basal and stress conditions.
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 1284. doi:1538-7445.AM2012-1284
Abstract 1295: Regulation of PAX2 activity by estradiol in breast cancer cells
Authors: Beauchemin David , Lacombe Catherine , Themsche Céline Van 
Published: August 2012

Abstract
Most breast cancer deaths arise from tumour cells ability to produce distant metastases. Among breast tumour subtypes, luminal A tumours, which express the highest levels of estrogen receptor alpha, are associated with the poorest metastatic phenotype and with longest relapse-free survival. Usually active during embryogenesis, PAX2, a homeobox transcription factor, is of a particular interest in this context. It is preferentially activated in luminal A tumours in vivo and it was shown to downregulate the expression of ERBB2 (erythroblastic leukemia viral oncogene homolog 2, HER-2/neu), a well-documented proinvasive and pro-metastastic gene, in these cells in response to estradiol in vitro. However, the ability of estradiol to regulate PAX2 activity and ERBB2 expression in non-luminal breast cancer cells, and the intracellular mechanisms involved in estradiol-induced PAX2 activity in breast cancer cells, have not been investigated. Objective: The objective of our work was to compare the impact of estradiol exposure on PAX2 activation in luminal and non-luminal breast cancer cells, and to determine the molecular mechanisms mediating estradiol-induced activation in breast cancer cells. Hypothesis: Estradiol would induce PAX2 activity in breast cancer cells of the luminal subtype only, in an estradiol receptor alpha-dependent manner. Results: Basal PAX2 activity was higher in cell lines from luminal compared to non-luminal subtype. Activation of PAX2 by estradiol, as assessed by measuring PAX2 phosphorylation on serine 393 residue and ERBB2 expression, was selectively achieved in breast cancer cell lines of the luminal subtype. This process was blocked by estrogen receptor alpha antagonist ICI 182780. Conclusion: PAX2 activation by estradiol is selectively achieved in breast cancer cells of the luminal subtype, via ERα. We are currently investigating intracellular signaling pathways downstream of estrogen receptor alpha which could mediate estradiol-induced activation of PAX2 in luminal 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 1295. doi:1538-7445.AM2012-1295