bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5181: Dynamic cellular adhesivity of breast cancer cells conferred by CD44 varies with mesenchymal or epithelial phenotype
Authors: Delgadillo Luis F. , Shirure Venktesh S. , Liu Tiantian , Goetz Douglas J. , Benencia Fabian , Burdick Monica M.
Published: August 2012

CD44 is a highly pleiotropic molecule whose many structural variants and glycan modifications determine function, particularly with respect to cell adhesion. In this study, we investigated the E-selectin ligand function of CD44 and its relationship with the mesenchymal versus epithelial cell phenotypes of breast cancer cells. Real time RT-PCR analysis for epithelial and mesenchymal cell markers showed that MDA-MB-231 and Hs-578T breast cancer cells are mesenchymal-like cells, but BT-20 and MDA-MB-468 breast cancer cells are epithelial-like cells. Flow cytometry revealed that all cell lines expressed CD44. Further exploration of the expression of specific CD44 isoforms at the mRNA level (by real time RT-PCR) and at the protein level (by flow cytometry) showed that mesenchymal-like breast cancer cells predominantly expressed CD44s but nearly undetectable levels of CD44v isoforms. On the other hand, epithelial-like cells expressed CD44s as well as CD44v isoforms. These results indicate that there are divergent CD44 isoform expression patterns in mesenchymal-like versus epithelial-like breast cancer cell lines. Further distinctions were observed in CD44 function in cells of the two phenotypes. Western blot analysis demonstrated that CD44 expressed by BT-20 cells were positive for E-selectin ligand activity, whereas CD44 of MDA-MB-231 cells failed to exhibit such functionality. Moreover, CD44v isoforms expressed by BT-20 cells, but not CD44s by MDA-MB-231 cells, mediated E-selectin adhesion under physiological flow conditions. Despite the positive ligand activity, none of the CD44v isoforms expressed by BT20 cells possessed HECA-452 reactivity by Western blot analysis. This finding indicates the absence of conventional sialofucosylated carbohydrates and the presence of potentially novel E-selectin reactive glycans. Collectively, these results show that CD44 expressed by epithelial-like cells, but not mesenchymal-like cells, are functional E-selectin ligands. Our investigation therefore implies an association of breast cancer cell adhesivity with the epithelial phenotype. Thus, future studies mechanistically linking the E-selectin ligand activity of CD44 isoforms with epithelial-to-mesenchymal transition may provide targets for novel breast cancer diagnostics, prognostics, and therapeutics.

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 5181. doi:1538-7445.AM2012-5181
Abstract 5267: Mouse models of cancer at The Jackson Laboratory Repository
Authors: Boswell Deborah M. , Rockwood Stephen F. , Sasner Michael , Lutz Cathleen M. , Davisson Muriel T. , Donahue Leah Rae
Published: August 2012

As one of the largest collections of characterized mouse strains available, The Jackson Laboratory Repository serves the scientific community as a resource to archive and distribute mouse models. Among the hundreds of new strains acquired or developed annually are a transgenic model for prostate hyperplasia that progresses to adenoma/adenocarcinoma, a transgenic model of large B cell lymphoma similar to tumors seen in patients with SLE and Sjogren's syndrome; and a targeted mutation strain of Lats1 (large tumor suppressor) that develops soft tissue sarcomas, ovarian stromal cell tumors and increased sensitivity to carcinogens. Conditional (floxed) strains that are useful in generating tissue-specific mutants have been recently added to the Repository and include mutations of Perp (TP53 apoptosis effector), Msh2 (mutS homolog 2, E. coli), and Irf8 (interferon regulatory factor 8). A transgenic strain that expresses Cre recombinase specifically in the large intestine has applications in colon cancer research. A combination mutant strain (containing targeted mutations of Vhl, von Hippel-Lindau tumor suppressor, and Pten, phosphatase and tensin homolog, and a transgene with Cre recombinase expression driven by Cdh16, cadherin 16) is a model for benign mixed adenosquamous genital tract tumors resembling clear cell cystadenomas observed in patients with Von Hippel-Lindau syndrome. The TEL-AML1 (Etv6-RUNX1, t12;21) translocation/fusion protein associated with childhood acute lymphoblastic leukemia is conditionally expressed by a targeted mutation strain. For transplantation research, a number of “humanized” mouse strains are available. These typically are immune compromised lines and/or lines that provide continuous circulation of human growth factors supporting a platform for a humanized microenvironment. A number of new research tool strains have been added to the Repository, including a fluorescent reporter for carcinoma metastasis, the Cre reporter R26R-Confetti strain which allows individual cell labeling with nuclear-localized membrane-targeted or cytoplasmic fluorescent proteins, and transgenic strains expressing the KikGR flurorescent protein which changes color from green to red upon activation. The Repository maintains a searchable on-line resource (www.jax.org) for each strain. Researchers can submit their strains by using the on-line form available at The Jackson Laboratory website: www.jax.org/donate-a-mouse. Supported by NIH, HHMI, The Ellison Medical Foundation and several private charitable foundations.

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 5267. doi:1538-7445.AM2012-5267
Abstract 5272: A novel organotypic 3D invasion platform to advance anticancer drug development
Authors: Tuomela Johanna M. , Seppänen Jani , Bernoulli Jenni , Salo Sirpa , Salo Tuula , Halleen Jussi
Published: August 2012

Tumor cell invasion is traditionally studied in three-dimensional (3D) organotypic models composed of type I collagen and fibroblasts, where various cell types and the extracellular matrix in the tumor microenvironment (TME) affects tumor cell behavior. As both molecular and cellular components of TME are cardinal participants in tumor progression, special attention should be addressed in the growth environment. As there is urgent need for more predictive models to be utilized in drug discovery and development, organotypic 3D in vitro assay provides an applicable platform. We characterized a novel organotypic model based on human uterine leiomyoma tissue to be used in drug development. To evaluate the general applicability of the myoma model, human tongue squamous cell carcinoma cells (HSC-3) and human triple negative breast cancer cells (MDA-MB-231(SA)-GFP) were cultured on top of human uterine leiomyoma tissue. Organotypic sections were examined by immunohistochemical stainings against cytokeratin and GFP, where invasion area and depth were measured. The organotypic myoma model also gives opportunity for ELISA, RIA and Western blotting analysis of the culture media. The used cell lines were found to invade in highly distinct patterns. This organotypic 3D myoma model provides a potent tool for analyzing the behavior of tumor cells. Moreover, we conclude that established organotypic 3D in vitro platform gives remarkable advantages to screen potential therapeutic options by producing totally new commercially usable models and innovative treatment approaches that target both the cancer cells and the TME components.

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 5272. doi:1538-7445.AM2012-5272
Abstract 5274: MicroRNA-214 /hepatoma-derived growth factor paracrine pathway contributes to hypervascularity of human hepatoma
Authors: Hsieh Sen-Yung
Published: August 2012

Background & Aims: Unusual hypervascularity is a hallmark of human hepatocellular carcinoma (HCC). Angiogenesis is essential for tumorigenesis and metastasis. While microRNA (miRNA) deregulation is implicated in carcinogenesis, the role of miRNAs in tumor angiogenesis remains unknown. Methods: MiRNAs deregulated in HCC were identified using array-based miRNA profiling. A luciferase reporter assay was used to confirm target association between miR-214 and hepatoma-derived growth factor (HDGF). Tube formation and in vivo angiogenesis assays were performed to validate the roles of miR-214 and HDGF in angiogenesis. Results: Downregulation of miR-214 was associated with HCC progression and poor clinical outcome. Ectopic expression of miR-214 suppressed cell proliferation and xenograft tumor growth and depressed the vascularity of the tumor and its surrounding tissues. The genes downregulated by ectopic expression of miR-214 were involved in the regulation of apoptosis, cell cycle, and angiogenesis. Integrated analysis implicated HDGF as a downstream target of miR-214. Conditioned medium of HCC cells contained bioactivity to stimulate tube formation of human umbilical vein endothelial cells, which was abolished by pretreatment of the conditioned media with HDGF antibodies, and silencing of HDGF expression or ectopic expression of miR-214 of the donor HCC cells. The angiogenic activity of the conditioned media lost by ectopic expression of miR-214 in the donor cells was restored by supplementation with recombinant HDGF. In vivo tumor angiogenesis assays showed significant suppression of tumor vascularity by ectopic expression of miR-214. Conclusions: A novel miR-214/HDGF paracrine pathway contributes to hypervascularity of HCC and is a candidate target for anti-HCC therapy.

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 5274. doi:1538-7445.AM2012-5274
Abstract 5276: Roles of BMP-9 signals during the formation of tumor vessels
Authors: Watabe Tetsuro , Suzuki Yuka , Yoshimatsu Yasuhiro , Miyazono Kohei
Published: August 2012

Angiogenesis plays critical roles during development and progression of cancer. Members of the bone morphogenetic protein (BMP) family are involved in TGF-beta superfamily, and have been implicated in the development and maintenance of vascular systems. While BMP-9 and 10 have been recently reported to bind to ALK-1 in endothelial cells, the roles of BMP-9/ALK-1 signaling in the regulation of endothelial cells have not yet been fully elucidated. Here, we examined the effects of BMP-9 on the proliferation of endothelial cells using various systems. Vascular tube formation from ex vivo allantoic explants of mouse embryos was promoted by BMP-9. BMP-9 also induced the proliferation of in vitro-cultured mouse embryonic stem cell-derived endothelial cells (MESEC) by inducing the expression of vascular endothelial growth factor receptor 2 and Tie2, a receptor for Angiopoietin-1. Decrease in ALK-1 expression and expression of constitutively active ALK-1 in MESEC abrogated and mimicked the effects of BMP-9 on the proliferation of MESEC, respectively, suggesting that BMP-9 promotes their proliferation via ALK-1. Furthermore, in vivo angiogenesis was promoted by BMP-9 in a Matrigel plug assay and a BxPC3 human pancreatic cancer xenograft model. In consistent with these in vivo findings, BMP-9 enhanced the proliferation of in vitro cultured normal endothelial cells from dermal tissues of adult mice and tumor-associated endothelial cells isolated from tumor xenografts in host mice. These findings suggest that BMP-9 signaling activates the endothelium via ALK-1. In this presentation, the roles of BMP-9 signals in the formation of lymphatic vessels will also be discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5276. doi:1538-7445.AM2012-5276
Abstract 5280: Activation of sphingosine kinase 1 is necessary for angiopoietin 2-induced hemangiogenesis and lymphangiogenesis
Authors: Yamada Akimitsu , Nagahashi Masayuki , Ramachandran Subramaniam , Kim Eugene Y. , Allegood Jeremy C. , Rashid Omar M. , Milstien Sheldon , Spiegel Sarah , Takabe Kazuaki
Published: August 2012

INTRODUCTION: Sphingosine kinase 1 (SphK1) is one of the two isoenzymes that generate the potent bioactive sphingolipid mediator, sphingosine-1-phosphate (S1P). S1P promotes cancer cell survival, proliferation and hemangiogenesis. The angiopoietin2 (Ang2)-Tie2 axis also plays a significant role in hemangiogenesis and lymphangiogenesis. The aim of this study was to investigate the involvement of SphK1 in Ang2-induced hem- and lymphangiogenesis. METHODS: Human umbilical vein endothelial cells (HUVECs) and human lymphatic endothelial cells (HLECs) were treated without or with an isozyme-specific inhibitor of SphK1 (SK1-I) and qPCR, western blot, ELISA, and tube formation assays carried out. We also used a new method we developed to quantify both hem- and lymphangiogenesis in the same sample in vivo by combining Directed In Vivo Angiogenesis Assays (DIVAA) with Fluorescence Activated Cell Sorting (DIVAA/FACS). RESULTS: qPCR revealed that human HLECs like HUVECs express high levels of SphK1 and the S1PR1 receptor. S1P rapidly stimulated xocytosis of Ang2 from HLEC storage granules determined by ELISA. Ang2 activated Tie2 in HLECs as demonstrated by its rapid phosphorylation on Tyr992, a major site responsible for its activation. Similar to Ang2, S1P also stimulated phosphorylation of Tie2, which may be due to secretion of Ang2. S1P is a potent hem- and lymphangiogenic factor for HUVECs and HLECs, respectively, as demonstrated by enhanced tube formation. Ang2-induced hem- and lymphangiogenesis was mediated at least in part by SphK1 as shown by the inhibitory effect of SK1-I. Furthermore, both S1P and Ang2 significantly induced hem- and lymphangiogenesis in vivo determined by DIVAA/FACS. Similar to its effect on in vitro hem- and lymphangiogenesis, inclusion of SK1-I in the angioreactors together with Ang2 completely abolished its ability to stimulate both of these processes. CONCLUSION: Activation of SphK1 is necessary for Ang2-induced hemangiogenesis and lymphangiogenesis through the SphK1/S1P and Ang2/Tie2 axes. This work was supported by NIH (K12HD055881) and Susan G. Komen for the Cure (KG090510) to KT, NCI (R01CA61774) to SS, and Sumitomo Life Social Welfare Services Foundation grant to MN.

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 5280. doi:1538-7445.AM2012-5280
Abstract 5288: Glioma stem cells stimulate the motility of brain endothelial cells: Identification of cell-adhesion molecules mediating motility and direct interaction
Authors: Burgett Monica E. , Tipps Russell S. , Lathia Justin D. , Bao Shideng , Rich Jeremy N. , Gladson Candece L.
Published: August 2012

Angiogenesis is a prominent characteristic of malignant glioma. Glioma stem cells (GSCs) reside adjacent to blood vessels in the perivascular niche, and recent evidence suggests a role for GSCs in promoting angiogenesis. Here we examined the interplay between GSCs and endothelial cells (ECs) on a laminin substrate, and the potential for GSC promotion of EC motility. In an in vitro 2D motility assay, we found that GSCs dramatically stimulated EC migration, and ECs also stimulated GSC migration. An antibody to integrin α6 blocked in part the motility of both GSCs and ECs. We also found that GSCs directly interact with ECs; therefore, we investigated which cell-adhesion molecules mediated the interaction. In a cell-cell binding assay, blocking antibodies to both L1-cell adhesion molecule (L1CAM) and integrin αvβ3 inhibited in part the direct contact between GSCs and ECs. In the motility assay, the combination of antibodies to L1CAM and αvβ3 significantly inhibited the motility and interaction of both cell types. These data suggest GSCs can promote angiogenesis by promoting EC migration, and indicate a role for the cell-adhesion receptors L1CAM and αvβ3 in the direct interaction between GSCs and ECs that may promote EC motility.

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 5288. doi:1538-7445.AM2012-5288
Abstract 5297: Autocrine stimulation of VEGFR-2 by leptin is associated with Notch signaling pathway and cancer stem cell marker expression
Authors: Guo Shanchun , Gillespie Corey , Liu Mingli , Gonzalez-Perez Ruben
Published: August 2012

Obesity-related cancer is an increasing health problem. Leptin, the major adipokine, has been suggested a major factor in these relationships. Leptin is a pro-inflammatory, pro-angiogenic and mitogenic factor for breast cancer cells. Recently, we described a comprehensive mechanism for leptin induction of VEGF, and unveiled a novel crosstalk between Notch, IL-1 and leptin (NILCO) in breast cancer. NILCO regulates VEGF/VEGR-2 in breast cancer cells and could represent the integration of developmental, pro-inflammatory and pro-angiogenic signals critical for leptin oncogenic actions. However, the specific mechanisms for leptin-induced VEGFR2 are unknown. To answer this question, we re-cloned mouse VEGFR-2 promoter and prepared VEGFR-2 luc, then analyzed VEGFR-2 regulation and expression by using several kinase inhibitors, leptin peptide antagonist 2 (LPrA2), a VEGFR-2 kinase inhibitor (SU5416), as well as siRNA against VEGFR-2, Notch1, Notch3 in 4T1 cells. Our results demonstrated that leptin-induced JAK2/STAT3, MAPK/ERK 1/2, PI-3K/AKT1, PKC, p38 and JNK signaling pathways may correlate with upregulated VEGFR-2. Moreover, leptin-induced activation of Notch/VEGFR-2 was linked to the upregulation of cancer stem cell markers CD44 and ALDH1. These leptin effects were inhibited by siRNA-Notch1&3, leptin signaling antagonist (PEG-LPrA2), VEGFR2-kinase inhibitor (SU5416) and siRNA-VEGFR-2. Our results show for the first time how leptin's molecular signals regulate VEGFR-2 (an autocrine survival process) that was associated to Notch and cancer stem cell marker expression. These novel findings may help to design new strategies for prevention/treatment of breast cancer through the abrogation of tumor angiogenesis and breast cancer stem cell renewal in an obesity context. [This work was supported in part by NIH/NCI1SC1CA138658-02; NIH/ARRA/3SC1CA138658-02S1 and the Georgia Cancer Coalition Distinguished Cancer Scholar Award (to RRGP)].

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 5297. doi:1538-7445.AM2012-5297
Abstract 530: Circulating γδ T cells are activated and depleted during progression of high-grade gliomas: Implications for γδ T cells therapy of GBM
Authors: Beck Benjamin , Gillespie G Yancey , Lopez Richard D. , Cloud Gretchen A. , Langford Cathy , Kim Hyung , Harkins Lualhati , Lamb Lawrence S.
Published: August 2012

Glioblastoma multiforme (GBM) remains frustratingly impervious to any existing therapy. We have previously shown that GBM is sensitive to recognition and lysis by ex vivo activated γδ T cells, a minor subset of lymphocytes that innately recognize autologous stress-associated target antigens via the T cell receptor and/or NKG2D. However, circulating γδ T cell counts and function are significantly reduced in patients with GBM. We sought to determine the role of γδ T cells in the immune response to high-grade gliomas in C57BL/6 mice bearing GL261 cell line-derived intracranial gliomas. Examination of mice 15 days after intracranial tumor placement and prior to development of neurologic symptoms revealed small infiltrating high-grade gliomas near the injection site. Concurrently, the peripheral blood γδ T cell count was significantly increased when compared to control mice that received only the methylcellulose vehicle (p=0.0003). T cell expansion was confined to the γδ T cell compartment, and a substantial proportion of γδ T cells in tumor-bearing mice expressed Annexin-V (20%-45%) indicating progression to apoptosis. Annexin-V expression in control mice was confined to 2-5% of γδ T cells, significantly less than tumor-bearing mice (p<0.0001). GL261-derived tumors expressed γδ T cell target stress-associated NKG2D ligands H60, MULT-1 and RAE-1, yet γδ T cells did not infiltrate the intracranial tumors. In a separate experiment, we found no survival advantage between wild-type (WT) mice over γδ T cell-deficient (C57BL/6α-/−) tumor-bearing mice. Based on this observation that activated γδ T cells did not invade the tumor, we treated mice with a stereotactic intracranial injection of 1.5 x 106 ex vivo expanded/activated γδ T cells approximately 15 minutes following placement of 5 x 106 GL261 glioma cells. Mice that received intracranial γδ T cells were protected from tumor development at site of injection, although distant tumors did eventually form. These data suggest that γδ T cells are stimulated in glioma-bearing mice and that depletion in γδ T cells observed in mouse glioma models and human GBM are likely due to activation-induced cell death in response to the tumor. Activated γδ T cells do not traffic to the tumor parenchyma; however, an antitumor effect can be achieved by direct placement of ex vivo activated γδ T cells in the vicinity of microscopic disease, suggesting a potential post-resection strategy for treatment of minimal residual GBM.

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 530. doi:1538-7445.AM2012-530
Abstract 5313: Sustained hypercalcemia primes non-invasive breast cancer cells for metastasis to high calcium microenvironments
Authors: Sakwe Amos M. , Koumangoye Rainelli B. , Ochieng Josiah
Published: August 2012

Breast cancer frequently spreads to calcium-rich organs/metastatic sites such as skeletal tissues but the role of hypercalcemia in breast cancer progression remains poorly understood. While patients with metastatic or end-stage cancer are likely to become hypercalcemic, up to 30% of breast cancer patients develop cancer-induced hypercalcemia (CIH) in the absence of metastases or bone diseases. The resulting high extracellular ionized calcium (Ca2+) activates the calcium sensing receptor (CaSR) that not only promotes the proliferation of breast cancer cells but also the secretion of parathyroid hormone-related protein by the tumor cells. This suggests that CIH may promote both the vicious osteolytic cycle and breast cancer progression. In this study, we examined whether sustained hypercalcemia primes breast cancer cells for metastasis to high Ca2+ microenvironments. We demonstrate that sustained hypercalcemia enables Ca2+-sensitive non invasive breast cancer cells such as MCF-7 to become high Ca2+-adapted and that this adaptation is accompanied by increased motility and more aggressive growth (increased proliferation). Consistent with this observation, invasive and metastatic breast cancer cells are high Ca2+-adapted cells that efficiently grow over a wide range of Ca2+ concentrations (up to 10 mM). Interestingly, we also show that at high Ca2+ (5 mM) breast cancer cells expressing the Q1011E CaSR variant that is common among African Americans proliferate faster than those expressing the A986S CaSR variant that is common among Caucasians. These data suggest that differences in the adaptation of breast cancer cells to hypercalcemia due in part to the expression of distinct CaSR variants underlie the distinct race-related breast cancer outcomes. This work is funded by the following grants made available to JO: DOD-W81XWH-07-1-0254 and 1SC1CA134018-01

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 5313. doi:1538-7445.AM2012-5313
Abstract 5376: Immunosuppressive T regs and MDSCs in human melanoma
Authors: Jordan Kimberly R. , McCarter Martin D.
Published: August 2012

Despite the ability to elicit active immune responses, most immunotherapies fail to eliminate tumors in human cancer patients. One reason immunotherapy has not met expectations may be due to immunosuppression in the local environment of the tumor. We have previously reported that untreated advanced-stage melanoma patients have increased numbers of peripheral T regs. In addition, exposure to melanoma cells in vitro expands T regs from normal human peripheral blood mononuclear cells (PBMCs). We have shown that melanoma-conditioned media have increased levels of TGF-beta, which may be responsible for the expansion of these suppressive cells in culture. Here we show that the number of peripheral T regs directly correlates with the amount of TGF-beta in the plasma of late-stage melanoma patients. We also show that melanoma patients have increased numbers of other suppressive cells, known as myeloid-derived suppressor cells (MDSCs). These MDSCs may further contribute to immunosuppression in advanced-stage melanoma patients. Finally, we show that the tumor-induced T regs and the T regs and MDSCs from the peripheral blood of melanoma patients suppress T cell responses in vitro, including proliferation, cytokine production, and cytotoxic activity. We are currently investigating whether MDSCs from melanoma patients contribute to the accumulation of T regs and whether immunotherapies used in the treatment of advanced-stage melanoma patients, particularly IL-2 and ipilimumab, affect their frequency or function. An understanding of the potential interaction between melanoma, MDSCs, and regulatory T cells will further our goal in identifying ways of blocking the function of these suppressive cells in melanoma patients to improve the effectiveness of future therapies.

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 5376. doi:1538-7445.AM2012-5376
Abstract 538: ROS differentially regulates prostate cancer cell survival
Authors: Chetram Mahandranauth A. , Bethea Danaya A. , Stowers Taylor J. , Hinton Cimona V.
Published: August 2012

In vitro and in vivo studies have shown that ROS accumulation correlates with increasing aggressiveness of prostate cancer cells, and increased expression of oncogenic molecules, such as CXCR4. CXCR4 is a G-Protein Coupled Receptor that is heavily implicated in the metastatic dissemination of prostate tumor cells to bone marrow. In cancer and other cell models, CXCR4 expression and activity increases in response to hypoxic environments and/or oxidative stress. Cells react by inducing expression of a HIF1α transcription factor, which enhances expression of targeted genes involved in adjusting to oxidative stress and cell survival. HIF1α has been shown to directly induce the expression of CXCR4 in prostate cancer cells. Therefore, generation of ROS in tumor tissues may critically influence CXCR4-mediated expression and functions, ultimately encouraging cancer progression. In this study, we elucidate a mechanism by which ROS induces CXCR4 expression and function in prostate cancer cells. We initially investigated the role of ROS on the phosphorylation of AKT (p-AKT) and ERK1/2 (p-ERK1/2) in DU145 and 22Rv1 prostate cancer cell lines. Using H2O2 as our model of ROS, we found that H2O2 induced a gradual expression of p-AKT and p-ERK1/2 by western blot analysis, which was abrogated by the ROS scavenger N-acetyl cysteine (NAC). It has been shown that PTEN is a negative regulator of AKT and ERK1/2 signaling, and that ROS accumulation inactivates PTEN phosphatase function; hence, we determined whether the increase in p-AKT and p-ERK1/2 expression was due to loss of PTEN function. By performing a phosphatase assay, we found that H2O2 inhibited PTEN catalytic activity compared to untreated cells, while NAC abrogated these effects in both cell lines. The increase in ROS and loss of PTEN has been shown to correlate with CXCR4-mediated function; therefore, we determined the role of H2O2 on CXCR4 expression. We found that H2O2 induced CXCR4 expression in DU145 cells, but not in 22Rv1 cells. Furthermore, we investigated whether ROS-mediated induction of CXCR4 was HIF1α dependent. Interestingly, H2O2 induced expression of HIF1α in DU145 cells, which was abrogated by siRNA for HIF1α. HIF1α expression decreased in 22Rv1 cells, which correlated with apoptosis, as confirmed by Annexin-V analysis. Finally, we observed that the H2O2-mediated increase in CXCR4 enhanced transendothelial invasion and migration in DU145 cells in a CXCR4 dependent manner. Our study suggests that ROS differentially regulates CXCR4 expression and function in prostate cancer cells, suggesting that ROS plays a differential role on heterogeneous tumor mass.

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 538. doi:1538-7445.AM2012-538
Abstract 5397: The SOCS-1 derived peptides inhibit tumor development
Authors: Scutti Jorge Augusto Borin , Matsuo Alisson , Massaoka Mariana , Figueiredo Carlos , Girola Natália , Valença Felipe , Melo Filipe Menegatti de , Travassos Luiz
Published: August 2012

Introduction: Melanoma is the most aggressive form of skin cancer and its incidence has increased dramatically over the years. The murine B16F10 melanoma in syngeneic C57Bl/6 mice has been used as a highly aggressive model to investigate tumor development. In our previous work, we demonstrate in the B16F10-Nex2 subclone that silencing of SOCS-1, a negative regulator of Jak/Stat pathway, leads to reversal of the tumorigenic phenotype and inhibition of melanoma cell metastasis. SOCS-1 silencing with shRNA affected tumor growth, cell cycle regulation with arrest at the S phase with large-sized nuclei, reduced cell motility and decreased melanoma cell invasion through Matrigel. In vivo, silencing of SOCS- 1 inhibited subcutaneous tumor growth and metastatic development in the lungs. Objectives: Since SOCS-1 is expressed in most melanoma cell lines and bears a relation with tumor invasion, thickness and stage of disease, we synthetized five SOCS-1 peptides based hydrophilic regions suggest regulating protein can be a target of cancer therapy. Suppressor of cytokine signalling 1 (SOCS 1) proteins are negative feedback regulators of several pathways involved in immune response, particularly the JAK/STAT (Janus kinase/Signal Transducer and Activator of Transcription. Results: No effect was found directly on B16F10 Nex 2 in vitro. In this work we hypothesized that SOCS-1 mimetic peptides should act on immune cells. Murine splenocytes from challenged animals and peptide treated was stimulated whole B16F10 Nex 2 lysate and supernatant was measure for ELISA to IFN-gama, IL-6, TNF-alfa and IL-10.The peptides increased the production of IFN-gama, IL-6, TNF-alfa and reduced IL-10 cytokine levels. Moreover, macrophages was stimulated and activation assay was performed. In vivo, the peptides are able to reduced significantly melanoma gorwth in metastatic and subcutaneous model.Additionally, mice knockout IFN-γ and CD1d revealed a inhibition of melanoma cell growth and metastasis. In the present study SOCS-1 derived peptides can be a target of cancer therapy.

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 5397. doi:1538-7445.AM2012-5397
Abstract 5469: Detection of deoxyguanosine adducts in the oral tissues of mice treated with the environmental carcinogen dibenzo[a, l]pyrene by LC-MS/MS
Authors: Zhang Shang-Min , Chen Kun-Ming , Aliaga Cesar , Gowdahalli Krishnegowda , Amin Shantu , El-Bayoumy Karam
Published: August 2012

Oral cancer is the major form of head and neck squamous cell carcinoma, which is the fifth most common cancer worldwide. Tobacco smoking is one of the leading causes of oral cancer. Dibenzo[a, l]pyrene (DB[a, l]P) is the most potent carcinogenic polycyclic aromatic hydrocarbon found in tobacco smoke. Our laboratory had developed a mouse model of oral cancer induced by DB[a, l]P. Our working hypothesis is that the stereochemical course of DB[a, l]P metabolism, the conformations of the DNA adducts formed and their removal by mammalian DNA repair enzymes, and their mutagenic properties if not removed in an error-free manner, all play critical roles during the induction of oral carcinogenesis. As an initial study to test our hypothesis, we have previously developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to detect and quantify DB[a, l]PDE-N6-dA adducts in oral tissues of mice treated with DB[a, l]P. We have shown that (-)-anti-cis- and (-)-anti-trans-DB[a, l]PDE-N6-dA adducts were detected from oral tissues of mice treated with DB[a, l]P. In the present study, to further test our hypothesis, we report on the development of a LC-MS/MS method to detect DB[a, l]PDE-N2-dG adducts in vivo. (±)-anti-[15N5]-DB[a, l]PDE-N2-dG adducts were synthesized as internal standards. The stereochemistry of adducts were characterized. Following the addition of internal standards, DNA isolated from oral tissues of mice treated with DB[a, l]P or DB[a, l]PDE was enzymatically hydrolyzed to 2′-deoxyribonucleosides and partially purified by solid-phase extraction. The LC-MS/MS analysis was carried out by monitoring transitions m/z 620 [M+H]+→ m/z 504 [(M+H)+−2′-deoxyribose] for DB[a, l]PDE-N2-dG adducts and m/z 625α m/z 509 for the internal standards. We have detected two N2-dG adducts from oral tissues of mice treated with DB[a, l]P, and four N2-dG adducts from oral tissues of mice treated with (±)-anti-DB[a, l]PDE. Collectively, the in vivo detection of dA and dG adducts indicated that DB[a, l]P is predominantly metabolized to (-)-anti-DB[a, l]PDE in oral tissues of mice. This sensitive LC-MS/MS method, is capable of simultaneously detecting both dA and dG adducts derived from anti-DB[a, l]PDE. Our results indicated that levels of dA adducts are significantly higher than dG adducts in vivo, which are consistent with those reported in literature in organs other than oral tissues, demonstrating that fjord region diol epoxide of DB[a, l]P predominantly form dA adducts than dG adducts.

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 5469. doi:1538-7445.AM2012-5469
Abstract 547: Mapping of estrogen receptor-dependent and independent signaling pathways for xeno- and phytoestrogens
Authors: Katchy Anne C. , Pinto Caroline , Bondesson Maria , Williams Cecilia
Published: August 2012

Xenoestrogens (e.g. bisphenol A) and phytoestrogens (isoflavones) are endocrine disrupting chemicals (EDC) that are capable of disrupting hormonal signaling in mammary and reproductive systems. These compounds are similar in structure to estrogen and bind to the estrogen receptors (ER). Estrogen receptors are involved in the regulation of cell cycle, proliferation, and apoptosis. It is likely that these compounds could affect the same biological processes or have their own subsets of target pathways. Here, we extensively identify target genes and signaling pathways for selected xeno- and phytoestrogens. Dose response curves for these compounds were measured using real-time PCR (RT-PCR) and transactivation assays of known estrogen target genes. Gene expression profilings for each EDC were performed on MCF-7 breast cancer cell lines using dual-color comparative array. Validations of array results were carried out using RT-PCR. Gene ontology and gene network mapping studies were done to identify the mechanism of action of these compounds. Non-ER mediated targets and the kinetics of each EDC were investigated in detail. Results show that many target genes regulated by bisphenol A and Genistein were involved in biological processes including cell cycle and proliferation, similar to that of estrogen. This contributes to an increased understanding of the mechanisms of EDC action, which may be implicated in development of diseases such as breast cancer and reproductive disorders, and would contribute to a more reliable risk assessment of different EDC compounds.

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 547. doi:1538-7445.AM2012-547
Abstract 5473: Integrative approach towards the functionality of CIAO1 gene
Authors: Lin Mau-Ting , Albertson Donna G.
Published: August 2012

Previous genome-wide analysis of copy number aberrations in oral squamous cell carcinoma (SCC) and oral epithelial dysplasia, a pre-malignant lesion, by array CGH has revealed that the genomes of these cancers and pre-cancers frequently harbor narrow (∼3Mb) regions of amplification on chromosome 2q11 region. Overexpression of CIAO1, a putative cytosolic iron-sulfur protein within this region, has been observed in patients’ cancerous samples and oral SCC lines. Knockdown of CIAO1 protein by RNAi results in growth retardation in oral SCC lines. Therefore, three HaCaT cell clones with an inducible Tet-on CIAO1 gene have been established to unravel the nature of the CIAO1 signaling network. Through the triangulation of protein-protein interactions, quantitative PCR array and transcription factor reporter array, we have investigated the early effects of CIAO1 overexpression in immortalized HaCaT cells, which may add to the understanding of oral cancer tumorigenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5473. doi:1538-7445.AM2012-5473
Abstract 5509: Liver cancer risk prediction model from a large prospective cohort in Taiwan
Authors: Wen Chi-Pang , Lin Jie , Yang Yi-Chen , Tsao Chwen-Keng , Etzel Carol , Huang Maosheng , Tsai Min-Kuang , Ye Yuanqing , Lopa Mishra , Hawk Ernest , Wu Xifeng
Published: August 2012

Models predicting liver cancer are only available for the high risk population with hepatitis carriers but not for the general population. we developed risk prediction models for liver cancer using data collected from a large health screening program in Taiwan with long term follow up. The cohort included 444,023 subjects at baseline. During a median follow up time of 7.4 years, 1,668 subjects developed liver cancer. Epidemiologic data, medical history and routine blood panel including serum transaminases data were collected with optional additional testing for HBV or HCV status. Liver cancer cases were ascertained by computerized record linkage with both the National Cancer Registry and the National Death Certification profiles in Taiwan. Stepwise Cox regression analysis was performed to identify significant predictors in the multivariate models. Individualized risk of developing liver cancer in 10 years was calculated from baseline probability and relative risk profile estimated from the Cox regression model. Models were developed separately to provide risk prediction for subjects who chose to have HCV tested (130,533 subjects at baseline and 416 liver cancer cases) and who chose otherwise (313,490 subjects and 1,252 liver cancer cases). Since the results for both sub-cohorts are comparable and we only reported results for the sub-cohort with HCV tested. In the sub-cohort with HCV tested, the model with only questionnaire data identified significant main effects for male gender, older age, prior history of diabetes, pack year of smoking, alcohol use, and physical inactivity. This epidemiologic model had an AUC of 0.798 (95% CI=0.772-0.815) for 10-year risk prediction. With only data from measures of Transaminases, the model achieved an AUC of 0.927 (95% CI=0.911-0.938), a significant increase as compared to the model with only epidemiologic variables. The addition of HBV improved the AUC to 0.936 (95% CI=0.913-0.958). Similar results were obtained for sub-cohort without HCV testing. The addition of HCV status in combination with HBV status, the AUC improve to 0.941 (95% CI=0.918-0.967). Among models using key history and blood panels, the use of serum Transaminases only, available in routine health check-ups, was able to provide most information needed in predicting liver cancer in the general public. Additional testing of HBV and /or HCV further increased the prediction power. This simple transaminase-based model for the general public can be valuable in clinical practice for identifying high risk individuals.

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 5509. doi:1538-7445.AM2012-5509
Abstract 5510: Assessing the level of breast cancer awareness among recently diagnosed patients in Ain Shams University Hospital
Authors: El-Shinawi Mohamed , Youssef AlMoataz , Aly Mohamed K. , Mostafa Mohamed , Alsara Mohamed , Yehia Ahmed , Hurlbert Marc , Mohamed Mona M. , El-Tawab Reda Abd
Published: August 2012

Introduction: 97% of Egyptian breast cancer patients are diagnosed at advanced stage, 70.6% possess positive lymph node metastasis on diagnosis while only 29.4% are free of metastasis reflecting late presentation of Egyptian patients. PURPOSE: Assess the level of knowledge about breast cancer among recently diagnosed breast cancer patients at Ain Shams University Hospitals. METHODS: In a cross-sectional, analytical observational study, we randomly enrolled 289 breast cancer patients with 45 of them who were recently diagnosed at Ain Shams University Hospitals meeting our inclusion criteria.Recurring patients or those unaware of diagnosis were excluded. Patients were interviewed using a specially designed questionnaire -of 75 questions over 60 minutes. Questionnaire gathered socio-demographic information, maternity history and tested knowledge of risk factors, signs and symptoms, screening options, management and prognosis of breast cancer was tested. Data was correlated to TNM staging. RESULTS: Patients’ age ranged between 30-72 years (mean 48±10.19). 40% were illiterate and only 15.6% finished higher education with 6-month average delay in seeking medical attention. Among the 45, TNM staging was assessed in 19 patients. Although 93% knew painless mass was symptom of breast cancer, 69% presented with painless mass and 47% knew someone with breast cancer prior to diagnosis, yet 24% thought they might have breast cancer. Regarding regular clinical breast exam 58% agreed to have it but 60% didn't know what was mammography, 56% didn't hear about breast self-exam and only 22% knew breast cancer had risk factors. Moreover, 58% thought treatment caused patients to be very ill, 29% thought breast cancer had good survival, yet 75% thought breast cancer is curable. Total breast cancer knowledge scores were low, with a median of 13 (out of 35) and mean of 12.64 ±5.12. CONCLUSIONS: Patients had acceptable awareness about breast cancer symptoms yet displayed gross ignorance about risk factors and poor awareness about screening, management and prognosis, correlating with their advanced TNM score. Therefore, raising breast cancer awareness and promoting education and early detection among our community is important.

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 5510. doi:1538-7445.AM2012-5510
Abstract 5527: Overexpression of protein phosphatase 4 is associated with poor prognosis in patients with pancreatic ductal adenocarcinoma
Authors: Weng Shaofan , Chen Weihong , Stanley Hamilton R. , Chatterjee Deyali , Katz Matthew H. , Lee Jeffrey E. , Pisters Peter W. , Gomez Henry F. , Abbruzzese James L. , Fleming Jason B. , Wang Huamin , Wang Hua
Published: August 2012

Background: PP4 has been reported to be overexpressed in breast and lung cancers. While previous studies have identified the role of PP4 in activating NFκB and JNK signaling pathway, the expression and functions of PP4 in pancreatic cancer has not been studied in detail. Methods: We examined the expression of PP4 protein in 133 patient samples of stage II pancreatic ductal adenocarcinoma (PDA) samples by immunohistochemistry (IHC). To confirm our IHC results, we measured the expression levels of PP4 mRNA in 15 PDA samples and 15 benign pancreatic tissue sample by real time RT-PCR. We also measured PP4 protein levels in in 9 fresh frozen PDA samples and their paired benign pancreatic tissues by Western blot. Using univariate and multivariate analysis, we correlated PP4 expression with survival and other clinicopathologic features. Results: PP4 was overexpressed in 75 of 133 (56%) stage II PDA samples by immunohistochemistry. Compared to benign pancreatic tissue, PP4 protein levels were higher in 9 of 9 PDA samples by Western blot analysis. PP4 mRNA expression levels in PDA samples were significantly higher than benign pancreatic tissue, suggesting that overexpression of PP4 in PDA is due to increase mRNA expression. PP4 overexpression was associated with higher frequencies of distant metastasis (p=0.02) and poor overall and recurrence-free survivals (p = 0.002 and 0.005) independent of tumor size, margin status, and lymph node status (stage) in patients with stage II PDAs. Conclusions: Our study shows that PP4 is frequently overexpressed in PDA samples and is associated with poor prognosis in patients with stage II PDA. Therefore, targeting PP4 signaling pathway may represent a new approach for the treatment of PDA.

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 5527. doi:1538-7445.AM2012-5527