bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4073: Comprehensive cell health analysis on a novel multi-parametric cell analyzer
Authors: Gillis Katherine , Khan Asima , Clor Julie , Tyagarajan Kamala 
Published: August 2012

Abstract
The study of anti-cancer compounds and cancerous cell lines requires access to cell health information that includes parameters such as cellular vitality, cell cycle and apoptosis and provides a complete picture of cell health. Traditionally, expensive and complicated instrumentation has been required for comprehensive cell health analysis. Here we present data from a novel, low-cost, easy to use, compact, high performance instrument, the Muse Cell Analyzer which provides multi-dimensional cell health information. The instrument utilizes a combination of highly simplified mix and read assay protocols with a touch-screen interface which greatly simplifies data acquisition and analysis. The objective of this study was to evaluate performance of Muse Count and Viability Assays when compared to traditional predicate methods including hemacytometers and automated image based systems. The mix and read format of the Muse™ Count & Viability assay provides quantitative detection of multiple suspension cell lines (e.g. Jurkats, Hl60s, K562s) and adherent cell lines (e.g. CHOs, PC3s, MCF-7s) across a wide range of concentrations and viabilities. Comparison of count and viability data from multiple platforms indicate that count and viability information on a vast variety of cancer cell lines can be robustly and accurately obtained on the Muse Cell Analyzer and the system depicts good linearity over a wide concentration range. Superior precision was obtained on the Muse Cell Analyzer for a variety of sample types with %CV ranging from 1-8% compared to %CV of 5-20% for predicate methods on the same samples. Cytotoxicity effects of anti-cancer compounds such as staurosporine, gambogic acid etc.on multiple cell types could be reliabily obtained on the instrument platform. The mix and read format of the Annexin V & Dead Cell assay and simplified Cell Cycle protocol on the instrument further allows for the easy and reliable study of impacts of anti-cancer compounds. The combination of three critical assays for cell count and viability, apoptosis and cell cycle status on a simple low cost instrument makes the Muse Cell Analyzer a versatile and accurate solution for comprehensive cell health analysis.
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 4073. doi:1538-7445.AM2012-4073
Abstract 408: Intermittent hypoxia effect on osteoclastogenesis stimulated by neuroblastoma cells
Authors: Bhaskara Vasantha Kumar , Mohanam Indra , Rao Jasti S. , Mohanam Sanjeeva 
Published: August 2012

Abstract
Neuroblastoma is the most common extracranial pediatric solid tumor that is derived from the developing sympathetic nervous system and results from the improper differentiation of neural crest cells. Neuroblastomas show a tremendous clinical heterogeneity ranging from benign ganglioneuromas to highly aggressive immature tumors. Hypoxia is a common feature of solid tumors and is associated with their malignant phenotype. Intermittent hypoxia (IH) which is characterized by cyclic periods of hypoxia and reoxygenation, occurs in tumor cells that are dependent on tumor blood vessels having intermittent perfusion fluctuations in blood flow. The occurrence of IH episodes varies significantly in rapidly growing malignant tumors. Bone is one of the target organs of metastasis in advanced neuroblastoma. To invade the bone, tumor cells produce osteoclast-activating factors that increase bone resorption by the osteoclast. The present study focuses on how IH-conditioned neuroblastoma cells modulate the differentiation of osteoclasts. IH-conditioned cells were derived by exposing tumor cells to 10 repeated cycles of hypoxia followed by reoxygenation. In order to assess the role of HIF-1α overexpression in osteolysis, we stably transfected human neuroblastoma cells with an expression vector containing a HIF-1α cDNA. Additionally, cells were stably transfected with a plasmid expressing HIF-1α shRNA or luciferase shRNA and selected in neomycin. The expression of HIF-1α protein was analyzed by western blotting in IH-conditioned and stable transfectants of neuroblastoma cells. Changes in gene expression were determined by real-time PCR for IL-8, MCP-1α, PTHrP, CXCR-4 and RANKL and the expression of osteoclastogenic factors was found increased in IH-conditioned and HIF-1α protein overexpressing cells. Conditioned medium collected from IH-conditioned and HIF-1α cDNA overexpressing neuroblastoma cells has shown enhanced osteoclast formation capabilities in the tartrate-resistant acid phosphatase (TRAP) assay. IH-conditioned and HIF-1α protein overexpressing neuroblastoma cells secrete increased amounts of VEGF protein, which has been found to promote osteoclast activity. Calcium-sensing receptor (CaR) plays a pivotal role in osteoclast differentiation. We found enhanced expression of CaR in RAW 264.7 (osteoclast precursors) cells treated with conditioned medium collected from IH-conditioned and HIF-1α protein overexpressing neuroblastoma cells compared with the control. Thus, IH was found to enhance osteolytic capabilities of neuroblastoma cells in vitro in part through HIF-1α protein stabilization.
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 408. doi:1538-7445.AM2012-408
Abstract 4090: Epigenetic profiling identifies differentially methylated loci in metastatic vs. non-metastatic primary squamous and basal cell skin cancers
Authors: Colacino Justin A. , Tang Alice , Moyer Jeffrey , Rozek Laura 
Published: August 2012

Abstract
Nonmelanoma skin cancer is the most commonly diagnosed cancer in the United States. Over 2 million new cases of these cancers are diagnosed each year, and incidence rates are increasing worldwide. Basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) of the skin are generally considered less aggressive than melanomas, which is reflected in their lower relative mortality rates. Despite this, a clinically relevant subgroup of both of these cancers will metastasize and develop an aggressive disease phenotype. In order to characterize the molecular pathways that may be involved in the development of metastatic disease in these common cancers, we measured DNA methylation profiles at 1505 CpG sites associated with 807 related to cancer using the Illumina Goldengate Methylation Cancer Panel BeadArray in BCCs and SCCs isolated from patients treated at the University of Michigan. DNA methylation was measured in metastatic (n=9) and non-metastatic (n=19) primary SCCs and in metastatic (n=3) and non-metastatic (n=9) primary BCCs. Due to the known divergent mechanisms of carcinogenesis of basal and squamous cell carcinomas, separate analyses were conducted for these tumor types. Unsupervised hierarchical clustering was used to identify overall differences in DNA methylation patterns between metastatic and non-metastatic disease. Gene specific analyses using linear modeling coupled with an empirical Bayes method of variance estimation were conducted to identify if methylation at individual CpG sites is associated with metastatic potential. SCCs were not found to cluster based on metastasis, however, metastatic BCCs were found to cluster together, suggesting that metastatic BCCs may share common mechanisms of pathogenesis. In gene specific analyses, FRZB was found to be statistically significantly hypermethylated in metastatic compared to non-metastatic primary SCCs (median methylation: 54.2% vs. 4.7%; p=3.8x10-5). Two sites in MYCL2 were found to be significantly hypomethylated in metastatic vs. non-metastatic BCCs (3.7% and 4.0% vs. 77.9% and 53.5%; p=1.87x10-6 and 3.69x10-5 respectively). The results from this pilot study give insight into the molecular mechanism of metastasis for nonmelanoma skin cancers and identify the divergent characteristics of BCCs and SCCs. The differentially methylation regions identified in this study may be useful epigenetic biomarkers of metastatic potential for nonmelanoma skin cancers to be examined in an extended patient population in future studies.
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 4090. doi:1538-7445.AM2012-4090
Abstract 410: NLRP3 is involved in tumor development by regulating immune function
Authors: Le-Bitoux Marie-Aude , Stamenkovic Ivan 
Published: August 2012

Abstract
The NLRP3 inflammasome is a multiprotein cytoplasmic complex that mediates the processing of pro-inflammatory interleukin-1β (IL-1β), IL-18 and possibly IL-33, via the activation of Caspase-1. While NLRP3 has been widely described in the pathogenesis of inflammatory diseases, its role in tumor-host interactions, where inflammation is known to be a major player, has yet to be fully elucidated. We observed that DMBA carcinogen treatment that induced large and numerous papillomas in the skin of wt mice induced barely visible lesions in NLRP3 deficient counterparts. Similarly IV injection of B16F10 melanoma cells resulted in multiple lung metastases in wt mice but virtually no metastatic growth in NLRP3-deficient animals. The NLRP3 inflammasome thus appears to be a key helper for tumor cell growth and survival at both primary and secondary sites. Bone marrow transplantation revealed that leukocyte-derived NLRP3 plays a key role in supporting tumor cell survival prompting us to compare the immune cell distribution and function in NLRP3 deficient mice to those in wt littermates, in both the presence and absence of tumor growth. Flow cytometry analysis of immune cell populations in these mice allowed us to link NRLP3 to the recruitment, adaptation and ability of the immune system to target tumor cells. We observed that compared to wt animals, NLRP3 KO mice had fewer CD3+CD4+ T cells, NK cells and F4/80+CD11c+ myeloid cells but more CD11b+Gr1+ cells, B lymphocytes and F4/80+CD206+ macrophages of the M2 phenotype in the spleen. The increased numbers of myeloid derived suppressor cells (MDSC) and M2 macrophages in mice that displayed decreased tumor growth was counterintuitive but was associated with functional impairment that could explain decreased permissiveness of the microenvironment for tumor growth. Our observations provide a framework to investigate the mechanistic implication of NLRP3 in the relationship between tumor cells and their microenvironment.
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 410. doi:1538-7445.AM2012-410
Abstract 4102: Histone deacetylase inhibitor, vorinostat, decreases transcription of survivin and is an enhancer of gemcitabine therapy for resistant pancreatic tumors
Authors: Green Leila J. , Koshkina Nadezhda V. , Curley Steven A. 
Published: August 2012

Abstract
Introduction: Overall 5-year survival rates of patients with pancreatic cancer remains below 15% due to delay in diagnosis and chemotherapeutic resistance. Combinational therapy has become the mainstay of treatment in patients who have metastatic pancreatic cancer. We hypothesize that 1) vorinostat (suberoylanilide hydoxamic acid), a histone deacetylase inhibitor, will improve efficacy of gemcitabine in gemcitabine-resistant pancreatic tumor cells; and 2) vorinostat works through decreasing the transcription inhibitors of apoptosis proteins (IAPs). Methods: Six different human pancreatic cell lines were treated with various levels of vorinostat with or without gemcitabine. Viability of tumor cells was determined via MTT assay and cytoplasmic levels of IAPs, including survivin, were analyzed via western blot assay. Real-time RT-PCR was performed on cell lysates to determine changes in survivin gene expression. Results: Combinations of vorinostat with gemcitabine significantly inhibited pancreatic cancer cells growth in vitro when compared with either treatment alone (p&lt;0.05). Screening of IAPs expression in tumor cells after exposure to vorinostat revealed variation of their expression in pancreatic tumor cells except survivin, whose expression declined consistently in all cells after vorinostat treatment. Overall cytoplasmic levels of survivin were seen to decline across all cell lines treated. Real-time RT-PCR showed a decrease in survivin gene expression in all cell lines tested as compared to control. Conclusion: We conclude that 1) vorinostat increases the efficacy of gemcitabine on pancreatic cells; and 2) decline of survivin protein levels in after exposure to vorinostat is due to a decrease in gene transcription and indicates that survivin plays an important role in improving the chemotherapeutic effect of gemcitabine. We are currently conducting in vivo experiments to verify our findings using orthotopic pancreatic tumor animal models.
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 4102. doi:1538-7445.AM2012-4102
Abstract 4107: Possible role of autophagy inhibition in hypoxia-induced chemoresistance of pancreatic cancer cells
Authors: Ko Yoon Ho , Won Hye Sung , Jeon Eun Kyoung , Hong Young Seon , Cho Young-Seok 
Published: August 2012

Abstract
BACKGROUND: Autophagy is a catabolic process and provides metabolic support for the cell by degradation of intracellular macromolecules. Various types of stress, including hypoxia, activate autophagy. Recent studies have suggested that hypoxia has been shown to associate with resistance to chemotherapy and radiotherapy and hence poor prognosis in pancreatic cancer. This study investigated the role of autophagy in the treatment of pancreatic cancer with gemcitabine under hypoxic condition. METHODS: To evaluate the role of autophagy inhibition in hypoxia-induced chemoresistance, BxPC-3 human pancreatic cancer cell line was used under normoxic and hypoxic conditions. We evaluated the extent of LC3-II, as an autophagosome marker, by Western blotting to measure the hypoxia- or chemotherapy-induced autophagy. We then examined the effects of gemcitabine on induction of apoptosis under normoxic and hypoxic conditions. Finally, to determine the effect of 3-MA, a known inhibitor of autophagy, on overcoming hypoxia-induced chemoresistance, the MTS assay and flow cytometry were performed.RESULTS: Compared with normoxia, gemcitabine-induced cell death was significantly decreased under hypoxia, as a result of the reduced apoptosis. Western blotting analysis demonstrated that LC3-II was increased under hypoxia, compared with normoxia. However, we found that 3-MA can enhance the growth inhibition and apoptotic effect of gemcitabine, even under hypoxia. These findings mean that autophagy mediates the chemoresistance under hypoxia. Conclusions: Activated autophagy plays a role in hypoxia-induced chemoresistance of pancreatic cancer cells. These findings may have important implications for future therapeutic strategies using gemcitabine against pancreatic 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 4107. doi:1538-7445.AM2012-4107
Abstract 411: PI3 Kinase gamma control of Arginase-1 expression promotes tumor immunosuppression
Authors: Schmid Michael C. , Foubert Philppe , Varner Judith A. 
Published: August 2012

Abstract
Tumor inflammation promotes angiogenesis, immunosuppression, and tumor growth. Tumor associated myeloid cells, which include incompletely differentiated myeloid derived suppressor cells, macrophages and granulocytes, establish an immunosuppressive microenvironment in tumors through the expression of Arginase-1, which depletes the tumor microenvironment of arginine, thereby promoting T cell death and NK cell inhibition. Suppression of Arginase-1 expression and function could alter the course of cancer therapy by stimulating anti-tumor immunity to work in concert with anti-tumor vaccine or chemotherapeutic agents targeting tumor cell proliferation and survival. We found that myeloid cells express Arginase-1 in response to IL-6 stimulation in a Jak2/Stat3 and PI3Kinase gamma/mTORc1 dependent manner by promoting the activation of the transcriptional enhancer C/EBPbeta. Pharmacological or genetic blockade of Jak2, Stat3, PI3Kgamma, mTOR or C/EBPbeta suppressed expression of Arginase-1. Suppression of PI3Kgamma or Arginase-1 expression blocked myeloid cell induced death of T cells in vitro. PI3Kgamma inhibition blocked Arginase-1 expression in vivo, thereby increasing the number of CD8+ T cells in tumors, stimulating T cell-mediated cytotoxicity of tumor cells in vivo and suppressing growth and metastasis of implanted and spontaneous tumors. These studies reveal that suppression of Arginase-1 expression with PI3Kgamma or Jak2/Stat3 inhibitors can serve as an effective therapeutic strategy in oncology.
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 411. doi:1538-7445.AM2012-411
Abstract 4110: Measurement of autophagy in sensitive versus resistant breast cancer cells
Authors: Shajahan Ayesha N. , Olson Rory , Webb Mathew , Samoy Jennifer , Zielinski John , Mullenix Michael , Clarke Robert 
Published: August 2012

Abstract
In 2011, there will be almost 192,000 newly diagnosed cases of invasive breast cancer in the U.S. with estrogen receptor positive ≤ positive (ER+) tumors. For these patients, endocrine therapy, administered as an antiestrogen, e.g., Tamoxifen (TAM) or Faslodex (Fulvestrant; ICI 182,780) or an aromatase inhibitor (AI), is the least toxic and most effective means to manage their hormone-dependent breast cancer. Tamoxifen (TAM) produces a 26% proportional reduction in mortality, however, advanced ER+ breast cancer largely remains an incurable disease and resistance to endocrine therapy remains a significant clinical problem. We and others have shown that antiestrogen resistant breast cancer cells have higher level of basal autophagy, a process that allows the cells to survive stress by digesting its own organelles. However, the precise role of this sustained autophagy in antiestrogen resistance is unclear and the measurement of autophagy flux, the dynamic measurement of passage of substrates through the autophagic pathway, is essential to determine levels of autophagy in a cell. In this study, we investigate the rate of autophagosome formation and degradation in antiestrogen sensitive versus antiestrogen resistant breast cancer cells. Autophagosome formation was measured using autophagosome specific fluorescent dye incorporation (Cyto-IDTM). Progression of autophagy was indicated by the degradation SQSTM1/p62 and NBR1 in cell lysates as measured by colorimentric enzyme immunoassays (ELISAs). The ELISAs were validated to provide biologically relevant sensitivity, dilutional linearity, reproducibility and recovery in native samples. Cells further progressing into apoptosis and cell death were identified by fluorescence using a combination of yellow emitting dye labeled Annexin V and a red emitting viability dye (GFP-Certified™Apoptosis/Necrosis detection kit). We hypothesize that combination of sustained autophagosome formation and inhibition of degradation are crucial in promoting pro-survival autophagy in antiestrogen resistant breast cancer cells. We show that while autophagosome formation and concurrent degradation of SQSTM1/p62 and NBR1 leads to cell death in sensitive cells, resistant cells have higher levels of autophagosomes, lower levels of degradation of SQSTM1/p62 and NBR1 and decreased cell death. Our data will highlight the importance of determining autophagy flux in pro-death versus pro-survival autophagy in drug resistance 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 4110. doi:1538-7445.AM2012-4110
Abstract 4116: Mammary adipocyte-specific metabolic alterations are associated with paracrine effects on mammary tumorigenesis
Authors: Volden Paul A. , Wonder Erin L. , Skor Maxwell N. , Carmean Christopher M. , Ye Honggang , Kocherginsky Masha , McClintock Martha K. , Brady Matthew J. , Conzen Suzanne D. 
Published: August 2012

Abstract
In rodent models of breast cancer, chronic social stress from isolated housing is associated with increased mammary tumor growth. Interestingly, in several diverse mouse strains, social isolation is also associated with increased expression of the key metabolic genes Acaca, Hk2 and Acly in the adipocyte fraction of the mammary gland. We hypothesized that these gene expression changes would be associated with increased mammary fat glucose metabolism, lipid synthesis, and secretion of paracrine growth promoting adipokines. We found that increased metabolic gene expression in mammary fat did indeed increase both lipid metabolism and leptin secretion from the mammary adipocytes of isolated mice. Surprisingly, visceral fat metabolism and systemic metabolic parameters were unchanged in social isolates, suggesting an effect of social stress that is specific to the mammary fat depot. When mouse mammary cancer cells were treated with culture media derived from the incubation of isolated versus group-housed mouse mammary adipose tissue, the mammary fat secretome of social isolates resulted in significantly increased cancer cell proliferation compared to mammary fat from grouped mice. Current experiments are designed to elucidate the metabolic factors promoting tumor cell proliferation in the mammary fat derived from the stressed animals. We propose a model wherein social isolation and the ensuing heightened neuroendocrine response significantly alters mammary adipocyte metabolism and associated adipokine secretion, thereby potentiating estrogen-independent mammary tumor growth. These studies have implications for understanding the environmental, endocrine, and molecular risk factors associated with estrogen receptor-negative breast cancer. DOD W81XWH-11-1-014901, NIH/NIDDK T32 DK087703, NIH R01CA148814
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 4116. doi:1538-7445.AM2012-4116