bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3998: Development of angiogenesis and cancer cell invasion assays in microfluidic flow cells for use with real-time microscopy
Authors: Conant Carolyn G. , Schwartz Michael , Ionescu- Zanetti Cristian 
Published: February 2011

Abstract
Understanding the molecular and cellular mechanisms of cancer progression from primary malignancy to metastatic disease is critical to development of successful treatments. Cell invasion from the primary tumor and tumor-induced angiogenesis are just two of many phenomena contributing to the pathology of metastatic disease. Both biological processes involve migration and transmigration of cells in response to chemoattractants. In vitro analysis of these phenomena typically involves deposition of a basement membrane derived matrix (Matrigel) in a transwell plate insert. This allows quantitation of either invasion or angiogenesis, but the inserts can be cumbersome to process. Here, we demonstrate a microfluidic based method to follow angiogenesis or cell invasion in real time. The method enables acquisition of high content data by microscopy using microfluidic flow cells which are optimized for imaging. The channels were filled with matrix in one half of the channel while the other half was maintained in the fluid phase. Using this technique, we tested endothelial cell response to VEGF and fumagilin. It was shown that VEGF impregnated in the matrix promoted angiogenesis while fumagilin abrogated the effect. We also investigated invasion of serum starved HT1080 and MCF-7 cells into FBS containing matrix and found that as predicted only HT1080 cells successfully invaded the matrix. Using these experimental conditions, the method is amenable to screening many cell-types or many simultaneous conditions or compounds by microscopy in the solid or liquid matrix of a microfluidic channel.
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 3998.
Abstract 4021: Genetic studies identify a map of redox signaling pathways in Drosophila
Authors: James Brian P. , Staatz William , Clark Elizabeth , Lu Jonathan , Powis Garth 
Published: February 2011

Abstract
Altered redox signaling, a consequence of oxidative stress and hypoxia, is seen in many human cancers and there is compelling evidence that altered redox signaling leads to tumorigenesis and aggressive tumor growth. In order to survive redox stress, tumor cells must initiate an adaptive response that facilitates both survival and drug resistance. The response to oxidative stress is largely restricted to tumor cells and thus offers a selective target for drug development. While it is clear that redox signaling is an important cellular response, our understanding of redox signaling pathways is rudimentary, and a map of redox signaling pathways comparable to that for protein phosphorylation is not available. In order to identify novel redox signaling genes we have undertook a classical dominant modifier screen in Drosophila melanogaster. Redox homeostasis is maintained in humans and Drosophila by scavenger systems such as superoxide dismutase (SOD), glutathione peroxidase and thioredoxin (Trx). In humans, Trx and glutathione are reduced and regenerated by thioredoxin reductase (Trxr) and glutathione reductase. Drosophila, and possibly insects in general, have only one glutathione/thioredoxin reductase gene encoded by the thioredoxin reductase-1 gene (Trxr-1). Thus, in Drosophila Trxr-1 serves as the starting point for all redox reactions in the cell and mutational disruption of the Trxr-1 gene leads to a genetically redox stressed animal suitable for screening. We screened a collection of second and third chromosome deficiency lines to identify genomic regions that dominantly suppress, or enhance the semi-lethal phenotype of Trxr-1 mutants. Our initial screen identified 18 genomic regions that dominantly suppress or enhance Trxr-1 mutants. We used overlapping deficiencies to validate and narrow down 9 genomic regions and have identified six individual genes with multiple alleles that show suppressing or enhancing phenotypes in four of these regions including mutations in two genes associated with lipid signaling and the Toll-like receptor, 18-Wheeler. The identification of 18-Wheeler indicates that the screen was successful, as 18-Wheeler, in addition to playing a critical role in the Drosophila immune response, is a member of the redox-regulated NF-κB pathway. This screen identified several genes not previously known to be involved in redox signaling, and begins to define a map of redox signaling pathways likely conserved through evolution.
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 4021.
Abstract 4067: A T-cell factor-4 motif regulates the phenotype of hepatocellular carcinoma cells
Authors: Koga Hironori , Walker Evan J. , Tsedensodnom Orkhontuya , Wands Jack R. , Kim Miran 
Published: February 2011

Abstract
T-cell factor (TCF) family proteins play a crucial role in regulating the activation of Wnt/β-catenin responsive target genes. Among the family proteins, TCF-4 is predominantly expressed in human HCC tissues. We have recently cloned 14 TCF-4 isoforms from human hepatoma cell lines, and naming them in alphabetical order from A to M. A TCF-4J and K pair has been identified based on the presence (K) or absence (J) of the SxxSS motif at the beginning of exon 9. This motif may modulate transcriptional activity of TCF-4 through phosphorylation/dephosphorylation of serine residues (Pukrop et al., Cancer Res 2001). However, cell-based functional analysis of this motif has yet to be performed. The AIM of this study was to investigate whether TCF-4J and K have distinct cellular phenotypes due to SxxSS motif expression. Methods: The human hepatoma cell line Huh-7 was used in this study. The cells were transfected with TCF-4J and K expression plasmids, and protein levels in cell lysates and nuclear extracts were examined by Western blot analysis. TCF-4K mutants with substitution of serine (S) to alanine (A) were generated by site-directed mutagenesis. Results: Protein expression of all cloned TCF-4 isoforms was confirmed by Western blot analysis. Expression levels of cyclin D1, which is a direct target of TCF-4, was increased in both cell lysates and the nuclear extracts of the TCF-4K, but not in TCF-4J-transfected cells. The TCF-4K isoform with a S269A mutation revealed a decrease in the expression level of cyclin D1, while the mutant with either a S272A or S273A substitution did not, which suggests that phosphorylation of serine 269 was critical to up-regulating cyclin D1 gene expression. Conclusion: The findings from functional analysis following mutagenesis of the SxxSS motif suggest that serine phosphorylation of this motif resulted in the altered target genes expression, including cyclin D1. Therefore, TCF-4 isoforms regulate the phenotype of HCC cells.
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 4067.
Abstract 4071: Prohibitin peptides induce apoptosis in cancer cells and inhibit angiogenesis
Authors: Rizwani Wasia 
Published: February 2011

Abstract
Prohibitin is a potential tumor suppressor protein that can repress E2F-mediated transcription and arrest cell proliferation. E2F1 binding domain of prohibitin is a coiled-coil structure and prohibitin functions through this domain to repress E2F1 transcription activity by recruiting HDAC1. The coiled- coil domain (175-217aa) by itself exhibited growth suppressive properties and showed a reduction in colony formation when transfected into breast cancer cells, T47D cells. It was shown that a synthetic peptide (42aa), designated as H1-6, corresponding to the coiled- coil domain induced apoptosis in four different human cell lines; MCF7, HAEC, BL18 and WI38 cells. Hence we decided to synthesize peptides of 14-mer (H1-2, H3-4, H5-6) and 7-mer (H2, H5) from the coiled coil domain of prohibitin and study their effect on cells. We found that each one of the prohibitin peptides (20μM) induced a marked increase in apoptosis in a panel of breast, lung and prostate cancer cell lines when compared to the control peptide (CP1). Interestingly, the peptides had no effect on breast epithelial cells, MCF10A. Treatment of MDA-MB-231 cells with 20μM of each prohibitin peptide individually reduced the invasive capability of these cells when compared to the control peptide. We also investigated their effect on angiogenesis in HUVEC cells and found that these peptides could significantly inhibit angiogenesis in vitro. In conclusion, these results suggest that these peptides could be of significant therapeutic value.
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 4071.
Abstract 4073: The PNA oligonucleotide is effective miRNA inhibitor that transfection reagents are not required
Authors: Oh Su Young , Ju Yeong Soon , Kim Seryun , Park Heekyung 
Published: February 2011

Abstract
The miRNAs are approximately ∼23 nucleotides non-coding RNAs. They regulate major processes such as development, apoptosis, cell proliferation, hematopoiesis, and patterning of the nervous system. MiRNA inhibition using antisense oligonucleotide is an effective tool for understanding miRNA function. The peptide nucleic acid (PNA) is an artificial oligonucleotide with a peptide backbone. PNAs have a stronger affinity and greater specificity than DNA or RNA and are resistant to nuclease, which is essential characteristic for a miRNA inhibitor that will be exposed to abundant serum and cellular nucleases. In here, we evaluated PNAs™ miRNA inhibitors in cultured cells including stem cell using luciferase assay and qPCR. FACS analysis was showed that PNAs™ miRNA inhibitors can penetrate to intra cells without transfection reagent. Also, miRNA inhibition effects are sustained until 15 days after treatment. PNAs™ miRNA inhibitor has a lower cytotoxicity than other nucleic acid-based miRNA inhibitors. PNAs™ miRNA inhibitors can be stored at room temperature for 20 weeks without losing the activity. These results demonstrate that PNAs are a powerful and effective tool to studies of miRNA mechanism.
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 4073.
Abstract 412: Isolation and analysis of metastasis-related genes in an oral cancer cells with autocrine stromal cell-derived factor −1/CXCR4 system
Authors: Uchida Daisuke , Onoue Tomitaro , Kuribayashi Nobuyuki , Tamatani Tetsuya , Nagai Hirokazu , Miyamoto Youji 
Published: February 2011

Abstract
We have previously demonstrated that distant metastasis might require the autocrine stromal cell-derived factor-1 (SDF-1; CXCL12)/CXCR4 system in cases of oral squamous cell carcinoma (SCC). In this study, we examined the expression of metastasis-related genes in B88-SDF-1 cells, which have an autocrine SDF-1/CXCR4 system and distant metastatic potentials in vivo. Microarray analysis revealed that 418 genes were upregulated in B88-SDF-1 cells. To confirm the specificity of the microarray analysis, 3 highly upregulated genes, metabotropic glutamate receptor 5 (mGluR5), plasticity related gene 1 (PRG1), and unknown gene were reanalyzed by RT-PCR. These 3 genes were upregulated in B88-SDF-1 cells in comparison with those in mock cells, but AMD3100, a specific antagonist of CXCR4, abrogated the upregulation of 2 (mGluR5 and PRG1) out of 3 genes in B88-SDF-1 cells. Thus, we analyzed the expression of metabotropic glutamate receptor 5 (mGluR5), one of the genes that were upregulated in B88-SDF-1 cells. A specific mGluR5 antagonist, MPEP did not affect the growth of B88-SDF-1 cells, but did inhibit the enhanced-migration of B88-SDF-1 cells. When the B88-SDF-1 cells were transplanted into the vein, numerous metastatic foci were detected. Under the present conditions, MPEP significantly inhibited the lung metastasis of the B88-SDF-1 cells, ameliorated body weight loss, and improved the survival rate of tumor-bearing nude mice. These results suggested that mGluR5 might be one of a possible metastasis-related gene induced by an autocrine SDF-1/CXCR4 system in oral SCC.
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 412.
Abstract 4192: Global gene expression profiling confirms the molecular fidelity of primary tumor-based orthotopic xenograft mouse models of medulloblastoma
Authors: Zhao Xiumei , Zhang Yujing , Baxter Patricia , Yu Litian , Voicu Horatiu , Gurusiddappa Sivashankarappa , Lau Ching C. , Leung Hon-chiu Eastwood , Li Xiao-Nan 
Published: February 2011

Abstract
Clinically relevant animal models are needed for understanding biology and testing novel therapies in medulloblastoma (MB), the most common malignant brain tumor that occurs in children. We have recently established a panel of orthotopic xenograft mouse models of MB through direct injection of fresh surgical specimens into the cerebella of Rag2/SCID mice. The xenograft tumors were previously shown to have replicated key histopathological phenotypes and invasive growth characteristics of the original patient tumors. The objective of the current study is to complete the molecular characterization of 11 MB models that have been serially passaged in vivo in mouse brains for at least three generations. Whole genome gene expression profiling was performed in duplicate with Illumina HumanWG6 v3 gene expression profiling beadchip that contains more than 48,000 probes. Our results showed that the xenograft tumors are molecularly accurate. When the nine passage I xenograft tumors were compared with their original patient tumors, high correlation coefficiency were observed (r2 &gt;0.95 in five models, &gt;0.9 in three models, and &gt;0.85 in one model). Serial subtransplantation up to five times did not cause significant changes of gene expression pattern (r2&gt;0.9 in eight models, &gt;0.8 in three models). Using RNAs from 4 normal fetal brains, one child cerebellum, and 5 adult cerebella as control, we further showed that these xenograft models, although molecularly independent displaying distinct gene expression profiles, can be classified into five different subcategories. A selected set of the differentially expressed genes were validated using real-time RT-PCR and Immunohistochemistry. In conclusion, this study provided strong experimental evidence to demonstrate that primary tumor-based orthotopic xenograft mouse models of MBs are molecularly accurate, and our findings of genetic abnormalities should significantly facilitate the rationally designed preclinical drug screenings in the near future.
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 4192.
Abstract 4315: Enrichment of xenotransplantable clonal cells in CD38high/CD138+ cells of multiple myeloma patients
Authors: Kim Dongkyoon , Weissman Irving L. 
Published: February 2011

Abstract
Multiple myeloma is a plasma cell neoplasm characterized by frequent relapse following initial therapy, implying the presence of a therapy-resistant and tumor-initiating cell population. While tumor-initiating cells have been reported for human multiple myeloma, the data provide conflicting information regarding the surface immunophenotype of these cells. Here, we analyzed bone marrow myeloma and B-lineage cells of patients for fractionation, examined engraftment of myeloma cells in various mouse models, and finally investigated myeloma-initiating population by examining the repopulation of human myeloma cells in recipient mice. The clonality of repopulating cells was also determined by RT-PCR of immunoglobulin gene variable regions. First, the examination of immunophenotype of patients’ bone marrow cells allowed us to fractionate myeloma/plasma cell-enriching CD138+/CD38high plasmacytic cells and CD19+CD38low/− B lymphocytic cells in 82% (31/38) of human samples. Then, we transplanted patient bone marrow cells into RAG2−/−γc−/−, NOD/scid or NOD/scid/γc−/− mice with/without human fetal bone pieces and examined the engraftment of myeloma cells. While human bone-bearing mice engrafted myeloma/plasma cells of 10 out of 19 samples, none of 18 samples gave rise to the repopulation of clonal myeloma/plasma/B cells in human bone-free immunodeficient mice. Further analysis of fractionated cells demonstrated that CD38high/CD138+ cells led to production of clonal human antibodies (2/10 samples) and a repopulation of clonally-related CD19+CD38low or CD138+CD38+ B lineage cells (5/10 samples) in grafted human bone of recipient mice. However, CD19+CD38low/−CD138− cells did not engraft clonally-related cells in human bone-bearing mice (0/9 samples). The analysis of CD38high/CD138+ myeloma cells demonstrated that more than 70% of patients still have heterogeneous plasmacytic cells in the expressions of CD45, CD19 or CD27. Our results suggest that human bone marrow microenvironment may be required for the survival and the engraftment of primary myeloma cells into recipient mice and that CD38high/CD138+/− plasmacytic cells but not CD19+CD38low/− B cells enrich clonal myeloma cells engrafting into recipient mice which may be responsible for tumor-initiation of multiple myeloma.
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 4315.
Abstract 4394: Development of parallel conjugation and assay methodologies to screen for antibodies with optimal properties for use as antibody-drug conjugates
Authors: Lyon Robert P. , Ryan Maureen C. , Sutherland May , Meyer Brad , Yu Changpu , Kostner Heather , Miyamoto Jamie , Benjamin Dennis R. 
Published: February 2011

Abstract
The activity of antibody-drug conjugates (ADCs) on cancer cells can be affected by a multitude of factors, such as binding affinity, rate of internalization, subcellular trafficking, and efficient drug release within the target cell population. Consequently, the properties of an ideal antibody for drug delivery are not necessarily the same as those for a therapeutic naked antibody. Furthermore, the use of indirect assays involving the use of secondary antibodies to screen for optimal ADCs can be misleading, since crosslinking on the cell surface can lead to altered downstream events, and the affinity of the secondary antibody constrains the dynamic range of the assay. When seeking candidate antibodies directed against a novel antigen for ADC therapy, it is therefore most desirable to screen a large panel in the form of ADCs and evaluate their cytotoxic activities, since these results provide a direct measurement of parameters that can affect cytotoxic activity. However, when dealing with microgram quantities of a large number of antibodies as is typical of an antibody discovery campaign, the yields from conventional conjugation methodologies are limiting. We developed a novel approach that addresses this issue and have successfully applied it to the discovery of optimal ADCs out of a large panel of candidate antibodies. The technology is currently being applied to the discovery of ADCs against antigens of interest for targeted drug delivery.
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 4394.
Abstract 4412: In vitro activity of the novel tubulin active agent BAL27862 in MDR1(+) and MDR1(−) human breast and ovarian cancer variants selected for resistance to taxanes
Authors: Duran George E. , Lane Heidi , Bachmann Felix , Sikic Branimir I. 
Published: February 2011

Abstract
BAL27862 is a synthetic small molecule that induces apoptosis due to the inhibition of tubulin polymerization via a potentially new mechanism of action. The activity of this novel compound was evaluated in human breast and ovarian cancer cell line variants selected for resistance by continuous exposure to taxanes. BAL27862 retained activity against MDR1 (ABCB1)/P-glycoprotein-mediated resistance variants, unlike vinca alkaloids and taxanes. Activity was also observed in the taxane-resistant ovarian 1A9/TxTP50 tumor cell line, which overexpresses TUBB3 and was approximately 35x resistant to taxanes. Moreover, BAL27862 retained activity in ovarian cancer cells with acquired mutations in the class I (M40) β-tubulin isotype (1A9/PTX10, Phe270-&gt;Val; 1A9/PTX22, Ala364-&gt;Thr) which confer taxane resistance. The MCF-7/TxTP50 breast tumor variant was 9x resistant to taxanes, and 5x resistant to epothilone B, yet retained sensitivity to BAL27862, the vinca alkaloids (vinblastine and vincristine), colchicine and anthracyclines. These cells have significantly reduced levels of the taxane-binding target, the class I β-tubulin isotype, and reduced levels of total α- and β-tubulin relative to wild-type cells, alterations which may be responsible for a 34% reduction in bound BODIPY-paclitaxel as detected by FACS analysis. Evidence of resistance to BAL27862 (3x) was only detected in the ovarian OVCAR-3/TxTP5 variant, which was also resistant to taxanes (8x), the vinca alkaloids (4x) and colchicine (2x) and ixabepilone (2x). Finally, silencing TUBB3 using a specific siRNA sensitized cells to taxanes but did not affect BAL27862 activity in both wild-type MCF-7 and OVCAR-3 cells relative to appropriate controls, suggesting that TUBB3 status may not influence BAL27862 response. Thus, BAL27862 is a new agent with a distinct in vitro activity profile against a broad panel of human breast and ovarian tumor cell lines resistant to standard tubulin-interacting agents. These data strongly support further development of BAL27862 as a novel anticancer agent.
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 4412.