bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract C46: Design, synthesis, and functional evaluation of leukocyte-function associated antigen-1 (LFA-1) antagonists in early and late stages of cancer development.
Authors: Sebastin Eider San , Zimmerman Tahl , Zubia Aizpea , Vara Yosu , Martin Elyette , Dejaegere Annick , Stote Roland H. , Lopez Xabier , Pantoja-Uceda David , Valcarcel Maria , Mendoza Lorea , Vidal-Vanaclocha Fernando , Cossio Fernando P. , Blanco Francisco J.
Published: June 2012

The integrin Leukocyte Function-associated Antigen-1 (LFA-1) recognizes and binds the Intercellular Adhesion Molecule-1 (ICAM-1) by its L chain Inserted domain (I-domain). This interaction plays a key role in cancer and other diseases. We report here the rational in silico design, small scale synthesis and biological activity evaluation of a novel family of potent LFA-1 antagonists. The structure-based design led to the synthesis of a family of highly substituted homoquiral pyrrolidines that mimic key residues of the ICAM-1 moiety. Some of these compounds showed both antiproliferative and anti-metastatic activity in a murine model of melanoma, as well as potent anti-adhesive properties in several cancer cell lines in the low micromolar range. The molecular mechanism of action of selected antagonists have been investigated by NMR analysis of their binding to the isolated I-domain of LFA-1. We show that the compounds bind to the I-domain Allosteric Site (IDAS), the binding site of other allosteric LFA-1 inhibitors. These results provide evidence of the potential therapeutic value of a new set of LFA-1 inhibitors, whose further development is facilitated by a synthetic strategy that is versatile and fully stereocontrolled

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C46.
Abstract C48: Detection of GNAQ/GNA11 mutations in circulating cell-free DNA as a biomarker of uveal melanoma.
Authors: Madic Jordan , Trouillet Benedicte , Gentien David , Milder Maud , Saada Stephanie , Assayag Franck , Nemati Fariba , Decaudin Didier , Desjardins Laurence , Piperno-Neumann Sophie , Lantz Olivier , Stern Marc-Henri
Published: June 2012

Background: Circulating tumor-derived DNA (ctDNA) can be detected in the plasma of cancer patients. Recently, ctDNA was proposed as a biomarker to follow tumor burden and monitor treatment efficacy (1). However, ctDNA represents a tiny fraction of the normal cell-free circulating DNA, and its assessment is challenging due to specificity, sensitivity and quantitative issues.

Patients and methods: We developed a real-time PCR based on the pyrophosphorolysis-activated polymerization (bi-PAP) for the quantification of ctDNA in plasma of patients with uveal melanoma (UM). Bi-PAP can detect known point mutations independently of large excess of normal DNA (2). Our assay targets the activating mutation of codon 626 of GNAQ or GNA11 which are present in most UM (3, 4).

Results: Bi-PAP primer pairs specific for GNAQ 626A>T, GNAQ 626A>C and GNA11 626A>T have been chosen. Their sensitivity and specificity were assessed on serial dilutions of tumor DNA in normal DNA. Each assay could detect a single mutated molecule per reaction, while 104 copies of normal DNA were not detected. In plasma of mice bearing UM xenografts, ctDNA were proportional to the amount of circulating human DNA and to the size of the xenograft. Furthermore, in a retrospective analysis of 19 UM patient sera, 8 were found positive.

Conclusion: Bi-PAP represents a promising tool for the quantification of ctDNA in the blood of UM patients. A prospective study is on going to confirm the clinical value of ctDNA level in metastatic UM patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C48.
Abstract C68: Suppression of AKT/mTOR antiapoptotic signaling by a novel drug candidate results in growth arrest and apoptosis of hepatocellular carcinoma-derived cell lines.
Authors: Cuconati Andrea , Mills Courtney , Zhang Xianchao , Yu Wenquan , Goddard Cally , Guo Haitao , Xu Xiadong , Block Timothy M.
Published: June 2012

Hepatocellular carcinoma (HCC) is the fourth most common cause of cancer fatalities worldwide, with chronic hepatitis B infection accounting for 60% of cases. In the majority of new diagnoses, primary tumor size contraindicates surgical resection, limiting available treatment options to rarely-curative interventions that generally have severe side effects. Overall, advanced HCC is largely incurable, with a five year survival rate of <5%. To expand the treatment pipeline, we conducted a high-throughput screening campaign of our 85,000-compound small molecule library to identify agents that would exhibit selective cytotoxicity for HCC-derived cell lines, as compared to immortalized non-transformed human hepatocytes. A disubstituted aminothiazole, termed HBF-0079, emerged as a potent and selective inhibitor of cell growth and viability in the HCC lines Huh7, Hep3b, HepaRG, MHCC97H and MHCC97L, and the hepatoblastoma line HepG2, with EC50 values from 0.7 to 33 μM. Conversely, growth and viability in the non-tumorigenic hepatocyte lines THLE2 and PH5CH, and several other non-HCC cancer lines, showed only moderate to no sensitivity. Subsequent structure-activity relationship studies have identified relevant features of the molecule, and resulted in two compounds that retain selectivity and improve EC50 to < 0.1 μM in Huh7. In a murine xenograft model, growth of Hep3B-derived tumors was significantly delayed by either systemic or intratumoral administration of HBF-0079. As determined in cell culture withdrawal experiments, incubation of Huh7 cells with HBF-0079 resulted in irreversible onset of growth arrest and apoptosis. Cell cycle analysis indicated arrest in G1/G0, with subsequent entry of cells into a sub-G0 population consistent with apoptotic cell death. While a putative molecular target is currently being confirmed, we have determined that treated Huh7 cells undergo loss of AKT phosphorylation at the mTORC2-targeted residue S473, with subsequent loss of phosphorylation of the mTORC1 targets SK6 and 4EBP1; these observations are consistent with cell cycle arrest and onset of apoptosis, ostensibly through loss of AKT survival signaling and global translational shutoff. Conversely, phosphorylation of Akt or mTORC1 targets is not affected in the non-sensitive PH5CH line. Thus, we have identified a cytotoxic molecule with in vitro selectivity for liver-derived cancer cells. In vivo efficacy studies and chemical hit-to-lead optimization are currently ongoing.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C68.
Abstract C96: A novel polyclonal antibody library for expression profiling of poorly characterized membrane and secreted human proteins.
Authors: Grifantini Renata Maria , Pagani Massimiliano , Pierleoni Andrea , Grandi Alberto , Parri Matteo , Campagnoli Susanna , Pileri Piero , Cattaneo Davide , Pontillo Angela , Canidio Elena , Nogarotto Renzo , De Camilli Elisa , Abrignani Sergio , Viale Giuseppe , Sarmientos Paolo , Grandi Guido
Published: June 2012

The YOMICS™ antibody library (http://www.yomics.com/) presented in this article is a new collection of 1559 murine polyclonal antibodies specific for 1287 distinct human proteins. This antibody library is designed to target marginally characterized membrane-associated and secreted proteins. It was generated against human proteins annotated as transmembrane or secreted in GenBank, EnsEMBL, Vega and Uniprot genome databases, described in no or very few dedicated PubMed-linked publications. The selected proteins/protein regions were expressed in E. coli, purified and used to raise antibodies in the mouse. The capability of YOMICS™ antibodies to specifically recognize their target proteins either as recombinant form or as expressed in cells and tissues was confirmed through several experimental approaches, including Western blot, confocal microscopy and immunohistochemistry (IHC). Moreover, to show the applicability of the library for biomarker investigation by IHC, five antibodies against proteins either known to be expressed in some cancers or homologous to tumor-associated proteins were tested on tissue microarrays carrying tumor and normal tissues from breast, colon, lung, ovary and prostate. A consistent differential expression in cancer was observed. Our results indicate that the YOMICS™ antibody library is a tool for systematic protein expression profile analysis that nicely complements the already available commercial antibody collections.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C96.
Abstract CN01-02: canSAR: An integrated cancer drug discovery informatics platform allowing systematic target evaluation.
Authors: Al-Lazikani Bissan , Patel Mishal , Bulusu Krishna , Halling-Brown Mark
Published: June 2012

Large-scale systematic analyses of the underlying genetics and proteomics of cancer, as well as the systematic mapping of effects of RNAi and chemical screens on different cancer cells are empowering cancer drug discovery and paving the way to wider stratified medicine approaches to the treatment of cancer. However, this deluge of data poses an equally large challenge of effectively integrating such massive and heterogeneous datasets in a manner that allows researchers to benefit from a multidisciplinary view across different. To address this need, we have developed a multidisciplinary cancer platform, canSAR (http://cansar.icr.ac.uk). It integrates millions of experimental datapoints ranging from chemical and RNAi screening, to expression, drug ADMET studies and 3D structural complexes. In this presentation we describe canSAR and show examples of how it has been utilized in the knowledge-driven prioritization of targets for drug discovery.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr CN01-02.
Abstract CN02-01: Functional genomics to identify targets and resistance.
Authors: Hahn William C.
Published: June 2012

Recent advances in genomics now make it possible to consider enumerating all of the genetic lesions in specific cancers. While these approaches will yield critical information regarding the identify, number, and types of alterations found in human tumors, a complementary approach to decipher the molecular basis of malignant transformation depends upon the application of genome scale tools to annotate the function of genes involved in cancer initiation and progression. Over the past several years, we have developed genome scale RNAi libraries and open reading frame expression libraries that permit a systematic evaluation of genes involved in cancer initiation and maintenance. Using these libraries, we have now performed screens in a panel of human cancer cell lines to systematically identify cancer vulnerabilities. By combining these functional approaches with information derived from mapping the structural abnormalities present in cancer genomes, we have identified several new oncogenes that contribute to cancer development. In pilot studies, we have identified several genes that act as synthetic lethal partners to known oncogenes or mediate resistance to targeted therapies. In recent work, we have expanded the scale of these experiments to genome scale in more than 100 cell lines, which has permitted greatly expanded and deeper investigation into genes that program the malignant state. Taken together, these studies suggest that combining forward and reverse genetic approaches with information derived from the cancer genome characterization projects will yield a comprehensive list of cancer vulnerabilities and establish a general approach for the rational identification of oncogenic and co-dependent pathways in cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr CN02-01.
Abstract CN05-01: Targeting Mdm2 in DNA repair and chromosome instability.
Authors: Eischen Christine M.
Published: June 2012

Mdm2, a direct inhibitor of the p53 tumor suppressor, is frequently overexpressed in many human malignancies. Increased levels of Mdm2 negatively impact genome stability, a common property of cancer cells. Recently, it has been appreciated that both p53-dependent and p53-independent functions of Mdm2 contribute to genome instability and tumorigenesis. Specifically, Mdm2 induces chromosome instability not only by inhibiting p53 activity, but also through inhibition of the Mre11/Rad50/Nbs1 DNA repair complex. We have shown that Mdm2 binds Nbs1 and inhibits early DNA damage signaling, resulting in a delay in DNA break repair. This delay in DNA repair leads to increased chromosome breaks, altered ploidy, and enhanced cellular transformation potential. Elevated Mdm2 levels also increase the severity and frequency of chromosome abnormalities that arise during aging and that have been linked to cancer development. Therefore, Mdm2 negatively influences genome stability, which contributes to tumorigenesis. However, this effect of Mdm2 on the genome may be therapeutically exploitable for the treatment of malignancies. In pursuit of this goal, we have begun testing small molecule inhibitors of Mdm2:p53 interaction, such as Nutlin, for consequences on DNA break repair.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr CN05-01.
Abstract CN06-02: TGF-beta and cancer stem cells.
Authors: Seoane Joan
Published: June 2012

Cancer stem cells also known as cancer-initiating cells (CICs) are cells with stem cell-like characteristics that are present within the tumor mass. CICs are considered to be responsible for tumor initiation, maintenance and recurrence, and hence are optimal therapeutic targets against cancer. Moreover, CICs are resistant to conventional therapy and, when inoculated in immunocompromised mice, generate tumors with the same characteristics as the original tumor from the patient. We have focused our studies in CICs obtained from human glioblastoma (GICs). Analyzing human glioma biopsies, primary cultured patient-derived tumor cells, patient-derived GICs and mice reproducing the human tumor, we have discerned the mechanisms and molecular determinants of GICs. The talk will be focused on the effect of TGF-beta on GICs. TGF-beta is a cytokine with an important role in cancer and several inhibitors of the TGF-beta signaling pathway are being developed to be used as anti-cancer therapies. However, it is crucial to understand the molecular pathways implicated in the malignant role of TGF-beta in oncogenesis in order to select the patient population that may benefit from an anti-TGF-beta therapy. We have observed that TGF-beta induces the self-renewal capacity of GICs and inhibition of TGF-beta decreases the amount of GICs present in the tumor. Our results have important implications on anti-TGF-beta therapeutic approaches that are being tested in clinical trials in our Hospital.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr CN06-02.
Abstract PL04-01: Sipuleucel-T and the active immunotherapy of prostate cancer.
Authors: Urdal David L.
Published: June 2012

Sipuleucel-T is an FDA approved autologous cellular immunotherapy indicated for the treatment of asymptomatic or minimally symptomatic metastatic castrate resistant (hormone refractory) prostate cancer. It consists of autologous peripheral blood mononuclear cells, including antigen presenting cells (APCs), which have been activated during a defined culture period with a recombinant human protein, PAP-GM-CSF, consisting of prostatic acid phosphatase (PAP), an antigen expressed in prostate cancer tissue, linked to granulocyte-macrophage colony stimulating factor (GM-CSF), an immune cell activator.

The development history will be summarized; the regulatory milestones will be described; and the lessons learned during the development of this novel approach to the treatment of prostate cancer will be discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr PL04-01.
Abstract PL05-03: Molecular basis and novel options for alternative epigenetic treatments against cancer.
Authors: Altucci Lucia
Published: June 2012

Deregulation of the epigenome is recognized as cause of cancer and epigenetic factors are receiving major attention as therapeutic targets; yet the molecular mode of action of existing epidrugs is largely elusive. Investigation of the spectrum of epimutations in cancer has only recently started and Food and Drug Administration-approved epi-drugs are currently limited by a number of factors. Mainly, the lack of knowledge about pathways and targets might have led to unfocused therapeutic applications of epi-drugs. We report here on the decryption of the mechanism of action of a novel epigenetic modifier, that inhibits multiple epi-enzymes. The rationale at the basis of targeting drugs could radically shift with the introduction of molecules interfering simultaneously with multiple targets as this might be more effective than single target agents thus representing a new generation of anti-cancer drugs. In this view, the inhibition of multiple epi-enzymes offers a promising alternative to combo-treatments of cancer using ‘poly-pharmacology’ approaches. Given the ability of cancer to bypass signaling routes via complex networks, the simultaneous modulation of multiple epi-targets promises to open new avenues against cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr PL05-03.
Abstract 1000: Inactivation of notch1 signaling suppresses FGFR1 knockdown-dependent growth inhibition of MDA-MB-231 breast cancer cells
Authors: Evangelou Andreas I. , Andrulis Irene L.
Published: June 2012

Breast cancer is a very heterogeneous disease as an array of expressed genes can be detected in individual tumors with varying phenotypic cellular morphology. However, despite this heterogeneity, two main histological subgroups predominate, basal-like and luminal-like tumors. Ongoing efforts aim to identify and target additional genetic alterations in breast tumors. One of the widely known and clinical distinctive markers between these tumor phenotypes for example is the transmembrane protein E-cadherin. Factors involved in the repression of E-cadherin are also involved in breast cancer progression. We have recently (Bane et al., Breast Cancer Research and Treatment 117:183-191, 2009) identified a set of differentially expressed and potentially additional distinctive markers between basal- and luminal-like breast tumors that have suggested an important role for Notch and FGF signaling in the development and maintenance of these breast cancer phenotypes. Other studies have also demonstrated gene amplifications in Notch and FGF receptors that can predispose in breast cancer pathophysiology. In the present study we have determined that downregulation of FGFR1 in the basal breast cancer cell line MDA-MB-231 decreased cell proliferation and survival. Interestingly, when Notch1 was also concomitantly repressed with siRNA, normal MDA-MB-231 cell proliferation and survival was restored. The current findings suggest and define a novel interplay between FGFR and Notch signaling in breast cancer patients that may be associated with poor prognosis due to ongoing and sustained basal-like phenotype in breast tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1000. doi:10.1158/1538-7445.AM2011-1000
Abstract 1002: Fatty acid synthase is overexpressed in Middle Eastern epithelial ovarian carcinoma (EOC) and its inhibition potentiates cisplatin-mediated apoptosis in EOC cells
Authors: Bu Rong , Jehan Zeenath , Ahmed Maqbool , Bavi Prashant , Hussain Azhar R. , Olyan Aishah Saad , Dayel Fouad Al , Ghourab Samir , Uddin Shahab , Kuraya Khawla Al
Published: June 2012

Fatty acid synthase (FASN), the enzyme responsible for de novo synthesis of fatty acids has been shown to be deregulated in several cancers including epithelial ovarian carcinoma (EOC). In this study, we investigated the function of Fatty acid Synthase (FASN) signaling pathway in a large series of middle eastern EOC patient samples, using a panel of cell lines, and nude mouse model. Using immunohistochemistry, we detected FASN in 75.5 %(114/151) tumor samples. Overexpression of FASN was significantly associated with tumor proliferative marker Ki-67 (p=0.0009), activated AKT (p=0.0117) and XIAP(p=0.0046). Treatment of EOC cell lines with C75 a selective inhibitor of FASN causes inhibition of EOC cell proliferation via induction of apoptosis. Inhibition of FASN by C75 led apoptosis via involving mitochondrial pathway, activation of caspases and downregulation of XIAP, and survivin. FASN inhibition causes down regulation of activated AKT. In addition, inhibition by FASN siRNA causes downregulation of FASN and activation of caspases suggesting the role of FASN in C75 mediated apoptosis. Furthermore, treatment of EOC cells with subtoxic doses C75 augmented the effect of cisplatin mediated inhibition of cell proliferation and induction of apoptosis. Finally, treatment of EOC cell line xenografts with combination of C75 and cisplatin resulted in growth inhibition of tumors in nude mice through downregulation of FASN and activation of caspases Altogether, our results suggest that overexpression of FASN in Middle eastern EOC suggests that FASN may be a potential therapeutic target in a subset of EOC, alone or in combination with other conventional chemotherapeutic agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1002. doi:10.1158/1538-7445.AM2011-1002
Abstract 101: Claudin-4 overexpression is associated with epigenetic derepression in gastric carcinoma
Authors: Kwon Mi Jeong , Kim Seok-Hyung , Jeong Hae Min , Kim Sung-Su , Lee Jae Eun , Gye Myung Chan , Koh Sang Seok , Park Cheol Keun , Shin Young Kee
Published: June 2012

The tight junction protein claudin-4 is aberrantly up-regulated in gastric cancer, but its clinical significance and the molecular mechanism underlying claudin-4 overexpression in gastric cancer remain unclear. Here, we investigated its roles and epigenetic mechanism regulating CLDN4 expression in gastric cancer. We show that increased membranous expression of claudin-4 in gastric carcinoma is associated with better patient prognosis, whereas cytoplasmic claudin-4 expression didn't show a significant association with prognosis. Consistent with the correlation of increased membranous claudin-4 with favorable clinicopathological factors, claudin-4 overexpression inhibited the migration and invasion of gastric cancer cells; in contrast, it didn't affect cell growth. Claudin-4 expression also increased the barrier function of tight junctions. Claudin-4 up-regulation was strongly correlated with DNA hypomethylation in both gastric tissues and gastric cancer cells. Moreover, loss of repressive histone methylations and gain of active histone modifications were associated with CLDN4 overexpression in gastric cancer cells. Interestingly, CLDN4 repression could be markedly derepressed by combined treatments that simultaneously target both histone modifications and DNA demethylation in CLDN4-hypermethylated cells, whereas concomitant changes in histone methylations and acetylations are required for CLDN4 induction in CLDN4-repressed cells with low DNA methylation. Taken together, this study reveals that membranous claudin-4 expression is associated with gastric cancer progression as well as it is an independent positive prognosis marker in gastric carcinoma. Furthermore, our findings suggest that epigenetic derepression may be a possible mechanism underlying CLDN4 overexpression in gastric cancer and that claudin-4 may have potential as a promising target for the treatment of gastric cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 101. doi:10.1158/1538-7445.AM2011-101
Abstract 102: Hypermethylation of Keap1 promoter and expression of the downstream genes in human colorectal cancer cell lines
Authors: Hanada Naoyuki , Takahata Takenori , Itoh Jugoh , Saijo Yasuo
Published: June 2012

Nrf2-Keap1 system is a master regulator of phase 2 detoxifying and antioxidant genes including NAD(P)H-quinon oxidoreductase 1[NQO-1] and aldo-keto reductase C1 (AKRC1). Somatic mutation of Keap1 gene and aberrant hypermethylation of Keap1 gene promoter region was found in human lung cancer. Loss of keap1 function activates Nrf2 activity and enhanced the lung cancer cell growth in vitro and in vivo. However, there has not been studied Keap1 gene status in colorectal cancer. In this study, we investigated status of keap1 gene in 10 human colorectal cancer cell lines. No somatic mutation of keap1 gene was found in all 10 cell lines. Hypermethylation of Keap1 promoter region was found in 8 cell lines. Keap1 mRNA and protein expression tended to be lower in these hypermethylated cell lines. The keap1 mRNA in a methylated cell line (HT29) was re-expressed by treatment with the DNA methylation inhibitor 5-aza-2’-deoxycytidine (5-azadC) and Trichostatin A (TSA). mRNA of the downstream genes, such as NQO-1 and AKRC1, were over expressed in the methylated cell line (HT29) compared to a nonmethylated cell line (Colo320DM) in Real time PCR analysis. In addition, we examined the methylation status of keap1 promoter region in primary tumors of 30 patients with colorectal cancer using methylation-specific PCR. An aberrant methylation in keap1 promoter was detected in 16 of 30 (53%). These findings suggest that hypermethylation of Keap1 promoter region in human colorectal cancer upregulates the expression of the downstream genes of keap1/nrf2 system which can be associated with the advantage for cancer progression and resistance to the anticancer agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 102. doi:10.1158/1538-7445.AM2011-102
Abstract 1022: TNFα-induced apoptosis is reduced, but inflammation is increased by combined exposure with common food additive carrageenan
Authors: Bhattacharyya Sumit , Dudeja Pradeep K. , Tobacman Joanne Kramer
Published: June 2012

Tumor necrosis factor (TNF)-α, a homotrimeric, pleiotropic cytokine, mediates both apoptosis and inflammation. Following exposure to TNF-α, activation of caspases leads to apoptotic cell death and stimulation of an inflammatory cascade through the non-canonical pathway of NF-κB activation leads to increased nuclear RelB and p52. In contrast, the common food additive carrageenan (CGN), which contains sulfated galactose residues, stimulates inflammation through both the canonical and non-canonical pathways of NF-κB activation. CGN-induced inflammation utilizes B-cell leukemia/lymphoma (BCL)-10 to activate a pathway of innate immunity that is initiated by interaction with toll-like receptor (TLR)-4. CGN also stimulates inflammation through a reactive oxygen species-mediated pathway. In a series of cell-based experiments, colonic epithelial cells and mouse embryonic fibroblasts were treated with TNF-α and CGN, in order to simulate the possible effects of exposure to dietary CGN in the setting of TNF-α-mediated effects. Marked increase in secretion of Interleukin (IL)-8 occurred, attributable to synergistic effects on phospho-NIK in the non-canonical pathway. TNF-α induced the ubiquitination of TNF receptor-associated factor (TRAF)-2 which interacts with NFκB-inducing kinase (NIK), and CGN induced phosphorylation of BCL10, which is required for NIK-phosphorylation. These results suggest that TNF-α and CGN in combination act to increase NIK phosphorylation, thereby increasing activation of the non-canonical pathway of NF-κB activation. In contrast, the apoptotic effects of TNF-α, including activation of caspase-8 and poly [ADP-ribose] polymerase (PARP)-1 fragmentation, were markedly reduced in the presence of CGN. CGN exposure caused reduction in Fas by cDNA microarray and by ELISA. The study results demonstrate that exposure to CGN drives TNF-α stimulated cells toward inflammation, rather than toward apoptotic cell death. The findings suggest that CGN exposure may compromise the effectiveness of anti-TNF-α therapy. Average consumption of CGN in the typical Western diet is reported to be 250 mg/day, in contrast to therapeutic glycosides, such as digoxin that are prescribed in doses of ∼0.1 mg/day. Ingestion of CGN-containing foods has the potential to both inhibit the pro-apoptotic effect and to increase the pro-inflammatory effect of TNF-α and may contribute to malignant transformation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1022. doi:10.1158/1538-7445.AM2011-1022
Abstract 1036: ProNGF/p75NTR signaling promotes survival of merlin-deficient vestibular schwannoma cells by activating NF-kB
Authors: Yue WeiYing , Ahmad Iram , Clark James Jason , Tryggvason Geir , Woodson Erika A. , Hansen Marlan R.
Published: June 2012

Vestibular schwannomas (VSs) represent benign Schwann cell (SC) neoplasms arising from the eighth cranial nerve and result from loss of function of the tumor suppressor gene, merlin. The mechanisms by which loss of merlin function promotes tumorigenesis in SCs is not well-defined. Denervation by axotomy leads to increased merlin inactivation by phosphorylation, increased p75NTR expression, JNK activation, and eventual death of normal SCs. In contrast to SCs, VS cells survive in the absence of axons. We found that p75NTR expression is elevated in sciatic nerves of mice harboring a SC specific dominant negative merlin mutation (P0SchΔ36-121) compared to nerves from wild-type mice suggesting that merlin status regulates p75NTR expression. Consistent with this we also find that p75NTR is over-expressed in primary human VS tissue compared with normal nerves. Treatment of normal rat SCs with proNGF, a high affinity p75NTR ligand, induces apoptosis by activating JNK. Primary cultured human VS cells, by contrast, display persistent JNK activity which promotes cell survival. Treatment of human primary VS cultures with the JNK inhibitors (I-JIP and SP600125) or JNK siRNA knock down results in decreased proliferation and increased apoptosis; treatment with proNGF protects cells from apoptosis due to JNK inhibition suggesting a p75NTR-mediated pro-survival signal in primary cultured VS cells. We explored the possibility that the pro-survival effect of p75NTR signaling results from activation of NF-κB. Indeed, proNGF treatment activated NF-κB in primary cultured VS cells while inhibition of JNK with SP600125 or siRNA-mediated JNK1/2 knockdown reduced NF-κB activity. Significantly, proNGF also activated NF-κB in primary cultured VS cells treated with JNK inhibitors. Thus, persistent JNK activity appears to be required for basal levels of NF-κB activity, but not for proNGF-induced NF-κB activity. To confirm that the increase in NF-κB activity by proNGF contributes to the prosurvival effect, we infected primary human VS cultures with Ad.IκB-SermutS32/36A virus which inhibits NF-κB activation. Compared to control virus, Ad.IκB-SermutS32/36A significantly increased VS cell apoptosis in VS cultures including those treated with proNGF. These results suggest that, in contrast to normal SCs which undergo apoptosis in response to p75NTR and JNK signaling, proNGF/p75NTR signaling provides a prosurvival response by activating NF-κB independent of JNK in merlin deficient VS cells. Such differences may account for the ability of human VS cells to proliferate and survive in the absence of axons.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1036. doi:10.1158/1538-7445.AM2011-1036
Abstract 1040: Downregulation of PI3K/Akt signaling is related to DHA-induced cytotoxicity in A549 human lung cancer cells
Authors: Kim Nayeong , Song Kyoung-Sub , Jing Kaipeng , Jeong Soyeon , Shin Soyeon , Oh Hyerim , Heo Jun-Young , Park Ji-Hoon , Seo Kang-Sik , Kwon Hyun-Joo , Park Jong-Il , Park Seung-Kiel , Kweon Gi-Ryang , Yoon Wan-Hee , Hwang Byung-Doo , Lim Kyu
Published: June 2012

Lung cancer is leading cause of all cancer deaths. Although omega-3 polyunsaturated fatty acids (ω3-PUFAs) have been reported to inhibit cell growth in several cancers, the anti-cancer mechanism of ω3-PUFAs on lung cancer is still unclear. In this study, we have investigated the mechanism of anti-cancer action of DHA, one of ω3-PUFAs, in A549 human non-small cell lung cancer (NSCLC) cell line. Following treatment of DHA, the viability of A549 cells was decreased in a dose-dependent manner. DHA induced apoptotic cell death as revealed by increased cleaved PARP, TUNEL positive cells and subG1 population. The amounts of PI3K and phospho-Akt proteins were decreased after DHA treatment dose-dependent manner. In addition, DHA decreased the level of phospho-phosphatase and tensin homolog deleted on chromosome ten (p-PTEN) protein, which is an inactive form of PTEN. Moreover, transient transfection of constitutive active Akt cDNA and full length of Akt cDNA into A549 cells partially restored DHA-dependent inhibition of cell growth compared with the cells transfected with kinase dead form of Akt. Taken together, these data suggest that inhibition of PI3K/Akt signaling pathway may be related to anti-cancer action of DHA in A549 human NSCLC cells. Therefore, utilization of DHA may represent a potential effective therapy for the chemoprevention and treatment of human non-small cell lung cancer. [This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (R13-2007-020-01000-0)]

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1040. doi:10.1158/1538-7445.AM2011-1040
Abstract 1043: Role of protein kinase Cι in alveolar rhabdomyosarcoma
Authors: Kikuchi Ken , Soundararajan Anuradha , Zarzabal Lee Ann , Weems Capella R. , Nelson Laura D. , Hampton Sheila T. , Michalek Joel A. , Rubin Brian P. , Fields Alan P. , Keller Charles
Published: June 2012

Background: Alveolar rhabdomyosarcoma (ARMS) is an aggressive pediatric cancer showing skeletal muscle differentiation. New therapeutic targets are required to improve the dismal cure rates for patients with metastatic ARMS. Protein Kinase C iota (PRKCI) has been shown to play an important role in many cancers but little to date is known of this kinase's role in rhabdomyosarcoma.

Materials and Methods: Human samples were obtained from the Pediatric Cooperative Human Tissue Network (Columbus, OH). Mouse samples were obtained from a Myf6Cre, Pax3: Fkhr, p53 conditional mouse model of ARMS and a Pax7CreER, Ptch1, p53 conditional model of embryonal rhabdomyosarcoma (ERMS). Expression of PKCι in tumors was evaluated by quantitative RT-PCR, western blotting and immunohistochemistry. To elucidate the functional role of PKCι in ARMS, we performed invasion assays, anchorage-independent colony formation assays and in vitro growth inhibition assays using primary cell lines obtained from mouse ARMS tumors, treating either with the PKCι specific inhibitor Aurothiomalate (ATM) or by PKCι RNA interference.

Results: Over-expression of PKCι at the mRNA and protein level was confirmed in human and mouse ARMS and ERMS. Immunoblotting confirmed that PKCι and its downstream targets, mitogen-activated protein kinase and nuclear factor-kappa B were expressed and highly activated in primary cell cultures. Inhibition of PKCι by RNA interference resulted in a dramatic decrease in cell viability and colony formation. Treatment of primary cell culture using ATM resulted in decreased cell viability (IC50: 54.3 – 256.9 uM) and colony formation, and increased G2/M blockade. Co-treatment with ATM and vincristine, a microtubule inhibitor currently used in rhabdomyosarcoma treatment regimens, resulted in a Combination Index of 0.572 – 1.017, suggesting cooperativity between these agents.

Conclusion: These results suggest that PKCι has an important function in proliferation of ARMS and that the PKCι specific inhibitor ATM shows activity in ARMS at clinically-feasible drug concentrations. Furthermore, these results suggest that combination therapy with vincristine and ATM should be investigated further in ARMS.

Ken Kikuchi and Anuradha Soundararajan contributed this presentation equally.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1043. doi:10.1158/1538-7445.AM2011-1043
Abstract 1051: Atypical PKC and TGFβ receptors co-operate to phosphorylate Par6
Authors: Gunaratne Adrian , Guglielmo John Di
Published: June 2012

Transforming growth factor beta (TGFβ) is involved in many aspects of cellular behavior including apoptosis and differentiation. In tumor cells, TGFβ signaling alters cell plasticity through a loss in apical-basal cell polarity and induces epithelial to mesenchymal transition (EMT). Integral to this process is the phosphorylation of a polarity protein, Par6, on a conserved serine residue (S345). Recent work has shown that phosphorylation of this site not only increases cell plasticity and EMT, but also promotes an invasive phenotype in breast cancer cells. Our work suggests that a polarity partner of Par6, atypical Protein Kinase C (aPKC), can also phosphorylate Par6 on this critical site to drive distinct cellular changes in non-small cell lung cancer (NSCLC) cells. Using co-immunoprecipitation studies and immunofluorescence microscopy, we show that aPKCι interacts with TGFβ receptors through the adaptor Par6, and that these proteins localize to the leading edge of migrating cells. Furthermore, co-expression of Par6 with TGFβ receptors results in increased phospho-Par6 levels as previously reported. However, we have also found that phosphorylation of Par6 increases in the presence of aPKC. aPKC kinase activity as well as association with Par6 were found to be important for Par6 phosphorylation as evidenced by a loss in phospho-Par6 using a kinase deficient mutant of PKCζ (PKCζ -KR) as well as a mutant of Par6 that does not bind aPKC (Par6-K19A). Furthermore, gene silencing of atypical PKC reduces TGFβ induced cell morphology changes, E-cadherin loss, and stress fiber production. Finally, gene silencing of atypical PKC was observed to reduce TGFβ dependent migration of NSCLC cells. In conclusion, our results indicate that atypical PKC cooperate with TGFβ receptors to regulate phospho-Par6 levels to drive an invasive phenotype of NSCLC 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 1051. doi:10.1158/1538-7445.AM2011-1051
Abstract 1055: Effect of embryonic stem (ES) cell-conditioned microenvironment on the reprogramming of melanoma
Authors: Kim Myoung Ok , Kim Sung-Hyun , Jeong Chul-Ho , Lee Kun-Yeong , Yao Ke , Bode Ann M. , Cho Yong-Yeon , Bowden Tim G. , Dong Zigang
Published: June 2012

The cancer micro-environment affects cancer cell proliferation and growth. Embryonic stem (ES)-preconditioned 3-dimensional (3-D) culture of cancer cells induces cancer cell reprogramming and results in a change in cancer cell properties such as differentiation and migration in skin melanoma. However, the mechanism has not yet been clarified. Using the ES-preconditioned 3-D micro-environmental model, we provide evidence showing that the ES micro-environment inhibits proliferation and anchorage-independent growth of SK-MEL-5 and SK-MEL-28 human melanoma cells, which is accompanied by the down-regulation of c-jun and c-fos gene expression. We also found that the ES micro-environment suppresses telomerase activity and thereby induces senescence in SK-MEL-5 and SK-MEL-28 cells. Furthermore, we observed that gremlin, an antagonist of bone morphogenic protein 4 (BMP4), is secreted from ES cells and plays an important role in cellular senescence. Taken together, these results demonstrated that gremlin is a crucial factor responsible for abrogating cancer properties in the ES-preconditioned 3-D micro-environment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1055. doi:10.1158/1538-7445.AM2011-1055
Abstract 1063: BRAFV600E status predicts higher uPA levels in papillary thyroid cancer
Authors: Darr E. Ashlie , Nowicki Theodore S. , Kummer Nicolas , MacEwan Melanie , Dunbar Julie , Suslina Nina , Iacob Codrin , Shin Edward , Schantz Stimson , Tiwari Raj , Geliebter Jan
Published: June 2012

Of the 44,670 new cases of thyroid cancer predicted for this year in the US by the American Cancer Society, 80% will be papillary thyroid carcinoma (PTC). PTC typically arises from a gain-of-function mutation in the RET, RAS, or BRAF genes which make up a linear signaling cascade for ERK activation in over 90% of PTC cases. PTCs possessing the BRAFV600E mutation have been associated with greater local invasion and regional metastatic potential. Urokinase plasminogen activator (uPA) has been shown to be an important mediator of invasion and metastasis in several cancers. To determine the extent to which BRAFV600E mutation status predicts up-regulation of the uPA in PTC, a study using patient thyroid tissue samples obtained over a 6-year period at a single tertiary care center was undertaken. DNA and RNA were obtained from patient PTC and matched, normal thyroid tissue samples using the Trizol method. BRAFV600E mutational status of the DNA was determined using the TaqMan SNP genotyping assay, while RNA was analyzed for differences in uPA transcription levels (relative to matched, normal thyroid tissue) by qRT-PCR. Fifty-four percent of the PTC samples possessed the BRAFV600E mutation. PTC samples bearing the BRAFV600E mutation displayed significantly higher uPA RNA levels (relative to matched control tissue) than those samples with wild-type BRAF (5.883 vs. 1.27-fold, p<0.05, Wilcoxon signed-rank test). Additionally, uPA RNA levels were significantly higher in patients with nodal metastasis (p<0.05). These data provide new evidence of the roles of BRAFV600E and uPA in PTC invasive pathology and demonstrate for the first time that BRAFV600E status is able to predict higher uPA levels in PTC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1063. doi:10.1158/1538-7445.AM2011-1063
Abstract 107: miR-200a targets SIRT1 for malignant transformation
Authors: Eades Gabriel , Yao Yuan , Zhou Qun
Published: June 2012

Evidence supports a critical role for microRNAs in regulation of tissue specific differentiation and development. Signifying a disruption of these programs, miRNA profiling has revealed extensive dysregulation in tumors compared with healthy tissue. The malignant contribution of some miRNAs is understood, however, in other instances the pathways affected by aberrant miRNA expression have yet to be characterized. The miR-200 family has been established as a key regulator of epithelial phenotype and as such, is deeply involved in EMT-like processes in cancer. While exploring a model of transformation of normal mammary epithelium we observed epigenetic silencing of the miR-200 family- as was previously identified in breast cancer cell lines. Here we show the class III histone deacetylase SIRT1 overexpression in a TGF-β driven model of transformation as well as in patient samples of early stage breast cancer. We establish SIRT1 transcript as subject to novel regulation of the miR-200 family, through miR-200a / miR-141 targeting. Moreover, we demonstrate that downregulation of miR-200 family members contributes at least in part to the overexpression of SIRT1, a proposed oncogene in breast cancer. Restoration of miR-200a or knockdown of SIRT1 results in decreased growth in soft agar and decreased invasion. These observations provide further evidence of the important tumor suppressive role of miR-200 family members in breast epithelium, as well as identifying a novel regulatory mechanism that may contribute to SIRT1 upregulation early in breast tumorigenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 107. doi:10.1158/1538-7445.AM2011-107