bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4254: DNA methylation profiling of ovarian tumors by methylation ligation-dependant macroarray.
Authors: Chelbi Sonia T. , Losi Lorena , Saponaro Sara , Martin Patricia , Braunschweig Richard , Benhattar Jean 
Published: August 2014

Abstract
Background: Ovarian cancer is the gynecological disease with the highest death rate. Efforts towards ovarian cancer research are needed to develop early detection tools, better therapies, cures and to prevent the disease. DNA methylation profiling of a large set of putative cancer biomarkers may help to identify relevant biological aspects of ovarian cancer.
Materials and Methods: We selected 106 patients diagnosed for ovarian cancer (35 serous, 30 endometrioid, 32 mucinous and 8 germinal tumors) for which clinicopathological informations were available and 17 histologically normal ovarian tissues as control. In order to identify methylation signatures, a method called Methylation Ligation-dependant Macroarray (MLM) and developed in our laboratory was applied to DNA extracted from microdissected formalin-fixed and paraffin-embedded tissue sections. The MLM technology, derived from a multiplex ligation assay coupled with an array-based analysis of uniform size amplification products, permits the methylation status analysis of 60 promoter regions at the same time.
Results: Each of the 106 ovarian tumor samples analyzed had a unique methylation profile. Hierarchical clustering identified mainly two groups of patients with distinct methylation profiles. The first group of tumors contained low levels of methylation, whereas the second group had higher methylation levels. The group 1 included normal ovarian tissues as well as a large number of ovarian serous tumors. Endometrioid and mucinous tumors were more represented in the group 2. When looking at the gene level, the highest methylation frequency was detected in the TAC1, SPARC, ADAMTS12 and hTERT promoter genes. The promoter of the BORIS/CTCFL cancer testis-antigen gene was found hypomethylated in several tumors, mainly in serous and germinal types. Located in the X-chromosome, the DAX1 promoter gene, which is partially methylated in normal ovarian tissues, was found hypomethylated or hypermethylated in tumors. Interestingly, the difference between the hypo- and hyper-methylated groups concerned mainly genes belonging to the WNT pathway.
Conclusion: The marked differences in methylation profiles independently of histological subtypes underscore the importance of characterizing tumors at the molecular level. Further investigations are needed to evaluate the role of the WNT signaling pathway as a marker and putative target for cisplatin chemosensitivity in ovarian tumors.
Citation Format: Sonia T. Chelbi, Lorena Losi, Sara Saponaro, Patricia Martin, Richard Braunschweig, Jean Benhattar. DNA methylation profiling of ovarian tumors by methylation ligation-dependant macroarray. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4254. doi:10.1158/1538-7445.AM2013-4254
Abstract 4259: Active hedgehog (HH) signal transduction in rhabdomyosarcoma (RMS).
Authors: Yoon Joon Won , Lamm Marilyn , Leong King-Fu , Iannaccone Stephen , Iannaccone Philip , Walterhouse David 
Published: August 2014

Abstract
RMS is the most common sarcoma of childhood. The majority of embryonal RMS (ERMS) and a minority of alveolar RMS (ARMS) express HH pathway components. However, a role for the HH signal transduction pathway is incompletely defined in RMS. Co-regulation of GLI1 and Patched1 (PTCH1) in ERMS and fusion-negative ARMS have suggested that the HH pathway is functionally intact in these subtypes. However, HH pathway activity has not been directly tested in RMS. Our aim was to demonstrate that some RMS cells have intact HH signaling pathways and are responsive to HH pathway activation or inhibition. We demonstrated expression of Indian HH, Desert HH, PTCH1, SMO and GLI1 by qRT PCR in Rh18 ERMS cells and Rh41 ARMS cells; Desert HH, PTCH1, SMO, and GLI1 in Rh 30 ARMS cells; and Indian HH, Desert HH, PTCH1, and SMO in RD ERMS cells. We confirmed high GLI1 protein expression by Western blot in Rh41 and Rh18 cells. Since HH signaling is believed to occur in primary cilia, we used immunofluorescence to demonstrate primary cilia in RMS-13 cells. Based on their differing levels of GLI1 expression (high in Rh41 and Rh18 cells and low/absent in Rh28 and RD cells), these RMS cell lines provided an opportunity to up- and down-regulate the HH pathway and measure effects on gene expression and cell function. Exposure of Rh41 cells to Sonic HH-containing conditioned media caused a 12-fold increase in PTCH1 expression and exposure of Rh28 cells to Sonic HH peptide increased GLI1 expression 1.5-fold, suggesting intact HH signal transduction in some RMS cell lines. We down-regulated HH pathway activity with the GLI1 inhibitor GANT61 and assessed Rh41 and RD cell viability by MTT assays. RMS cell viability decreased following exposure to increasing concentrations of GANT61. The reduced cell viability appeared to result at least in part from down-regulation of the anti-apoptotic GLI1 target gene, BCL2. These results suggest a role for HH signaling in RMS cell survival. Understanding the role of HH signaling in RMS biology may contribute to successful introduction of HH pathway inhibitors into RMS treatment.
Citation Format: Joon Won Yoon, Marilyn Lamm, King-Fu Leong, Stephen Iannaccone, Philip Iannaccone, David Walterhouse. Active hedgehog (HH) signal transduction in rhabdomyosarcoma (RMS). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4259. doi:10.1158/1538-7445.AM2013-4259
Abstract 4262: The role of SPP1 in NPC microenvironment.
Authors: Chen Jinna 
Published: August 2014

Abstract
Nasopharyngeal Carcinoma (NPC) is highly associated with EBV infection and lymphocytes infiltration, which contribute to form the NPC microenvironment. To better understand the NPC microenvironment, we conducted PCR array using pooled cDNA mixtures from 5 NPC tissues and their counterpart non-tumor tissues to profile inflammation-associated gene expression. We found that the Th1/Th2 related cytokines were down regulated in NPC tissues. This result was verified by Real-time quantitative PCR using 13 pairs of NPC samples and non-tumor tissues. The result of IHC staining of NPC tissues with T helper cell marker CD4 also showed decreased infiltration of T helper cells. Lower level of Th1/Th2 related cytokines were also detected in NPC serum compared to paired non-tumor serum. Osteopontin (SPP1), a glycophosphoprotein, playing roles in inflammation, bone formation, and tumor progression and metastasis, was over expressed in 67% (8/12) of NPC samples compared with non-tumor tissues. IHC staining of SPP1 in NPC tissues also showed up-regulation of SPP1 in either NPC cells or lymphocytes. To study the effect of SPP1 on T helper cells population, we cultured PBMC by adding SPP1 protein at the concentration from 0 to 100 ng/ml into the culture medium, followed by Flow Cytometry analysis. We found that Low-dose SPP1 protein stimulated T helper cell proliferation, which could be inhibited by SPP1 at high dose. Our data indicated that the over-expression of SPP1 by NPC cells might contribute to tumor immunosuppression in NPC microenvironment, thus promote cancer progression.
Citation Format: Jinna Chen. The role of SPP1 in NPC microenvironment. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4262. doi:10.1158/1538-7445.AM2013-4262
Abstract 4268: Resistance to growth inhibition by transforming growth factor-beta is associated with activation of the PI3K-Akt pathway in HPV16-immortalized human keratinocytes at late stages of in vitro progression.
Authors: Velidandla Rupa , Vaswani Gehana , Kowli Sangeeta , Pirisi-Creek Lucia , Creek Kim E. 
Published: August 2014

Abstract
Transforming growth factor-beta (TGF-ß) plays a vital role in embryonic development, cell proliferation, inflammation and wound repair. The Smad proteins are the central mediators of TGF-ß signal transduction. However, recently several non-Smad pathways (ex., Erk, Jnk, p38 MAPK, PI3K-Akt) have been determined to play key roles in TGF-ß signaling. TGF-ß signaling is often disrupted in cancer, including cervical cancer. Human papillomavirus (HPV) is the main etiological agent in cervical cancer. HPV type 16 (HPV16)-immortalized human keratinocytes (HKc/HPV16) are initially very sensitive to growth inhibition by TGF-ß. However, HKc/HPV16 become increasingly resistant to TGF-ß during in vitro progression, and are completely resistant to TGF-ß-induced growth inhibition at late-stages of transformation, when they exhibit a differentiation resistant phenotype (HKc/DR). Although HKc/DR are completely resistant to the growth inhibitory effects of TGF-ß, some Smad signaling remains. A reporter construct containing tandem repeats of Smad binding elements was induced by TGF-ß in HKc/DR at about 50% of the level observed in HKc/HPV16. Furthermore, TGF-ß was capable of inducing epithelial-mesenchymal transition (EMT) in HKc/DR. To further explore TGF-ß signaling in HKc/HPV16 and HKc/DR, we used phospho specific antibody microarrays from Full Moon BioSystems. These arrays feature phospho and non-phospho pairs of specific antibodies pertinent to the TGF-ß pathway. Protein extracts from HKc/HPV16 and HKc/DR untreated or treated for 90 min with 40 pM TGF-ß were compared on the antibody microarrays. Activation of the Akt pathway by TGF-ß was observed in HKc/DR but not in HKc/HPV16. Similarly, PI3K and mTOR showed activation by TGF-ß in HKc/DR but not HKc/HPV16. We conclude that TGF-ß signals primarily through Smad-dependent pathways to inhibit cell proliferation in HKc/HPV16, but that non-canonical (non-smad) signaling through the PI3K-Akt-mTOR axis predominates in HKc/DR.
Citation Format: Rupa Velidandla, Gehana Vaswani, Sangeeta Kowli, Lucia Pirisi-Creek, Kim E. Creek. Resistance to growth inhibition by transforming growth factor-beta is associated with activation of the PI3K-Akt pathway in HPV16-immortalized human keratinocytes at late stages of in vitro progression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4268. doi:10.1158/1538-7445.AM2013-4268
Abstract 4278: Mapping deregulated pathways downstream of RTKs in cell line models.
Authors: Rikova Klarisa , Hall Benjamin , Possemato Anthony , Mroszczyk Keri , Lee Kimberly A. , Ren Jian Min , Guo Ailan , Mulhern Daniel , Wang Yi , Beausoleil Sean A. , Comb Michael J. 
Published: August 2014

Abstract
In virtually all epithelial tumors, growth factor receptor activity is deregulated by activated mutations, genomic amplification, and autocrine loops. The dependence of tumor cell survival upon the driving oncogene has been called “oncogene addiction” and demonstrates the acute sensitivity of cancer cells to inhibition of the pathways driving their proliferation, growth and survival. Major questions regarding the sensitivity of solid tumors to targeted kinase inhibitors is why some tumors respond and others do not and why ones which respond still are able to develop resistance through the activation of other pathways. Robust, but not durable, response to receptor tyrosine kinase inhibitors (RTKIs) highlights the need to inhibit pathway activity at multiple levels.
In this study, we identify extensive signaling networks downstream of receptor tyroine kinases (RTKs) across multiple spaces, including phosphorylation, acetylation, and methylation. We used the established method of TMT labeling coupled with serial peptide immunoprecipitation applied to profile drug-treated and untreated RTK pathway-addicted cancer cell lines. Quantitative analysis was performed to look at the effects of drugs on RTK-dependent cell lines: MKN45 (Met) with Crizotinib, H3122 (Alk) with Crizotinib, H1703 (PDGFRα) with Gleevec, and H3255 (EGFR) with Iressa. Phosphotyrosine, acetyl-lysine, methyl arginine, ATM/ATR substrate (s/tQ), and AGC/CAMK/STE family kinase motif were all selected for use in peptide immunoprecipitation which allowed us to characterize and quantify a broad PTM space. In this study we were able to identify and quantify hundreds of modification sites for phosphorylation, arginine methylation, and lysine acetylation. Those sites found to be drug-sensitive, including all PTM types, were used for pathway analysis of signaling downstream of each RTK disease driver.
Citation Format: Klarisa Rikova, Benjamin Hall, Anthony Possemato, Keri Mroszczyk, Kimberly A. Lee, Jian Min Ren, Ailan Guo, Daniel Mulhern, Yi Wang, Sean A. Beausoleil, Michael J. Comb. Mapping deregulated pathways downstream of RTKs in cell line models. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4278. doi:10.1158/1538-7445.AM2013-4278
Abstract 4288: RASSF1C, unlike RASSF1A, reduces TNF-α induced phosphorylation of MST1/2.
Authors: Amaar Yousef G. , Firek Matthew , Reeves Mark E. 
Published: August 2014

Abstract
Introduction: Recently, RASSF1A has been shown to mediate the apoptotic effects of TNF-α by interacting with the mammalian sterile 20-like kinase 1 and 2 (MST1/2) through the Salvador/Rassf/Hippo (SARAH) domain leading to MST1/2 phosphorylation activation of apoptosis through the Hippo pathway. In contrast, we have previously shown that RASSF1C promotes cell proliferation and attenuates apoptosis in cancer cells. Since both RASSF1A and RASSF1C contain the SARAH domain located in their identical C-termini, RASSF1C should be capable of interacting with SARAH domain-containing proteins, and could potentially attenuate MST1/2-mediated apoptosis through the Hippo pathway. Thus, in this study we have investigated the impact of RASSF1C on MST1/2 activation/phosphorylation in presence of TNF-α, including whether RASSF1C modulates MST1/2 pro-apoptotic effects through the Hippo pathway differently from RASSF1A.
Method: Breast and lung cancer cells over-expressing RASSF1A and RASSF1C were used in this study. Cells were cultured in the proper media and treated with TNF-α for 12-18 hr. Cells were collected and used for Western blot analysis utilizing antibodies that detect MST and p-MST antibodies.
Results: Our preliminary data suggest that RASSF1C over-expression decreases the phosphorylation levels of MST1/2 in TNF-α-treated breast and lung cancer cells compared to those cells over-expressing RASSF1A
Conclusion: Our findings suggest that over-expression of RASSF1C in breast and lung cancer cells may attenuate the MST1/2 apoptotic effects by sequestering of MST1/2 proteins and inhibiting their activation by phosphorylation. The findings also further support our hypothesis that RASSF1C attenuates apoptosis; and RASSF1C, unlike RASSF1A, may inhibit MST1/2 activation and hence may negatively modulate the Hippo pathway.
Citation Format: Yousef G. Amaar, Matthew Firek, Mark E. Reeves. RASSF1C, unlike RASSF1A, reduces TNF-α induced phosphorylation of MST1/2. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4288. doi:10.1158/1538-7445.AM2013-4288
Abstract 4304: PPP1R1B-STARD3 fusion gene promotes gastric tumorigenesis through activation of PI3K/Akt signaling.
Authors: Yun Sun Mi 
Published: August 2014

Abstract
Fusion genes result from genomic aberrations such as translocation, amplification and rearrangement. They have the potential to be new targets for the diagnosis and therapeutics in several different types of cancers. To identify fusion genes in gastric cancer, we conducted RNA sequencing with 18 gastric cancer cell lines and 11 pairs of primary human gastric cancer tissues and their adjacent normal tissues. We identified PPP1R1B-STARD3 fusion gene and validated the expression of this fusion gene by Real-Time PCR in the expanded panel of the human gastric tissue samples. Interestingly, PPP1R1B-STRAD3 fusion gene was detected in approximately 30% of primary human gastric cancers, but not in adjacent matched normal gastric tissues. Overexpression of PPP1R1B-STARD3 in MKN-28 significantly increased cell proliferation and colony formation. This increased proliferation was mediated by activation of PI3K/Akt signaling. To confirm if the expression of PPP1R1B-STARD3 fusion gene affects the tumorigenic capacity of gastric cancer cells in vivo, we performed the xenograft assay with MKN-28 cells that stably express PPP1R1B-STARD3 fusion gene. Overexpression of the fusion gene enhanced the MKN-28 tumor growth in nude mice. Hence, PPP1R1B-STARD3 fusion gene overexpression gives growth advantage in vitro and in vivo. Taken together, these findings indicate that PPP1R1B-STRAD3 fusion gene increased cell proliferation through the activation of PI3K/Akt signaling in gastric cancer cells.
Citation Format: Sun Mi Yun. PPP1R1B-STARD3 fusion gene promotes gastric tumorigenesis through activation of PI3K/Akt signaling. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4304. doi:10.1158/1538-7445.AM2013-4304
Abstract 4309: Fibulin-2 suppresses tumor growth and angiogenesis through the inhibition of Erk1/2 and p65 pathways in nasopharyngeal carcinoma.
Authors: Shuen Wai Ho , Lung Maria Li 
Published: August 2014

Abstract
Nasopharyngeal carcinoma (NPC) has distinctive ethnic and geographic distributions, with the highest incidence in Southern China. Infection with the human herpesvirus, Epstein-Barr virus, genetic predisposition, dietary, and environmental factors are high-risk factors for the development of NPC. Evidence indicates that loss of heterozygosity on chromosome 3 is one of the early events contributing to the cancerization of normal nasopharyngeal epithelial cells. By using a chromosome 3 NotI genomic microarray, Fibulin-2 (FBLN2) mapping to chromosome 3p25.1, was previously identified as a candidate tumor suppressor. FBLN2, which is an extracellular matrix (ECM) protein, contains four domains, regions consisting of cysteine-rich and cysteine-free, anaphylatoxin-like domain, serial EGF-like repeats, and a fibulin module. FBLN2 is silenced by promoter hypermethylation in NPC. Re-expression of FBLN2 in NPC cell lines significantly reduces colony formation, cell proliferation, migration, invasion, and in vivo tumor formation and angiogenesis. In this study, the aim is to investigate the molecular mechanisms and signaling pathways of FBLN2-mediated tumor suppressive and anti-angiogenesis effects. Full-length FBLN2 was subcloned into inducible tetracycline-regulated and lentivirus systems. Phosphorylated forms of the key signaling molecules, Erk1/2 and NFκB, were found to be significantly reduced in FBLN2 stably-expressing NPC cell lines after phosphorylation target screening using Western blot analysis. FBLN2 also induces changes in the cytoskeleton arrangement, possibly through the regulation of Rho GTPases. Thus, the activities of Rho GTPases were determined by the Rho pull-down assays and immunofluorescence staining with confocal microscopy. FBLN2 is able to reduce the tumor formation ability via inhibiting Erk1/2 and NFκB pathways and to inhibit the cell migration and invasion via regulating Rho GTPase activities. FBLN2 is a secretory protein and is generally silenced in NPC, suggesting the possibility that it may be useful as a diagnostic marker and/or new target for therapeutics of NPC. Therefore, these studies provide strategic new insights into the understanding of the mechanistic role of FBLN2 in NPC and enhance our understanding of the molecular genetic basis of NPC tumorigenesis.
Citation Format: Wai Ho Shuen, Maria Li Lung. Fibulin-2 suppresses tumor growth and angiogenesis through the inhibition of Erk1/2 and p65 pathways in nasopharyngeal carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4309. doi:10.1158/1538-7445.AM2013-4309
Abstract 4320: Agtuzumab, a humanized monoclonal antibody, blocks AGR2 function through conformational epitopes around its catalytic center.
Authors: Li Dawei , Wu Zhenghua , Zhu Qi , Guo Hao , Gao Guangwei , Mashausi Dhahiri Saidi , Chen Hao 
Published: August 2014

Abstract
AGR2 is a member of protein disulfide-isomerase (PDI) family and plays an important role in cancer development including tumorigenesis and metastasis. Unlike the classical PDI protein with a CxxC catalytic center, AGR2 contains a CxxS center that function in Mucin2 formation on the endoplasmic reticulum. However, AGR2 has also been shown to present in blood plasma and is secreted by several cell lines. As a secreted small protein, AGR2 is particularly suitable target for antibody medicine and its functional antibody is under investigation for therapeutic potentials by several major pharmaceutical firms and academic institutions but few showed AGR2 any anti-cancer efficacy. Here we report a humanized monoclonal antibody against AGR2, Agtuzumab, with promising therapeutic potentials. This antibody blocks AGR2 function in cell migration and proliferation assays and reduced tumorigenic and angiogenic potential of xenograft tumors in nude mice. Treatment of AGR2-blocking antibody reduced the expression of VEGF and VEGFR level in xenograft tumor tissues for both human tumor and its mouse stroma cells. Scanning mutagenesis assays revealed discontinuous regions around the CxxS PDI center. on AGR2 protein that are essential for efficient binding of Agtuzumab. Our results for the first time confirmed with a potential therapeutic antibody that AGR2 can be a therapeutic target and its blocking requires the blockage of a conformational epitope. This result also explains why many monoclonal antibodies developed against peptides may not be functional against AGR2 related tumors.
Citation Format: Dawei Li, Zhenghua Wu, Qi Zhu, Hao Guo, Guangwei Gao, Dhahiri Saidi Mashausi, Hao Chen. Agtuzumab, a humanized monoclonal antibody, blocks AGR2 function through conformational epitopes around its catalytic center. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4320. doi:10.1158/1538-7445.AM2013-4320