bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5031: R-spondin potentiates Wnt/β-catenin signaling in Ewing sarcoma cells.
Authors: Pedersen Elisabeth A. , Scannell Christopher A. , Lawlor Elizabeth R. 
Published: August 2014

Abstract
Proper regulation of Wnt signaling is highly context-dependent and critical for growth and cellular maintenance. It is well-established that dysregulation of the canonical Wnt/β-catenin pathway contributes to the development and progression of numerous malignancies. Despite this, there has been little evidence to support a role for canonical Wnt signaling in Ewing sarcoma (ES) pathogenesis. Recent studies have shown that when members of the leucine-rich G-protein coupled receptor (LGR) family, namely LGR4 and LGR5, are bound by their ligands, R-spondins, they potentiate Wnt/β-catenin activity. Importantly, recent studies from our lab have shown that some ES tumors and cell lines express high levels of LGR5. In addition, R-spondin is highly expressed in the developing bone, which is the primary site of ES in children. Since ES cells do not exhibit canonical Wnt/β-catenin activity in standard culture conditions, we hypothesized that cues from the tumor microenvironment may be required for activation of this pathway. In particular, we hypothesized that R-spondins would potentiate Wnt activity in LGR-expressing ES cells. To test this hypothesis, ES cells were cultured in the presence of WNT3A and/or R-spondin 2 (Rspo2). Immunocytochemistry was used to determine β-catenin nuclear localization, and Wnt transcriptional activity was determined by flow cytometry and luciferase activity using cell lines transduced with lentiviral 7xTCF-GFP and 7xTCF-luciferase reporters, respectively. qRT-PCR was used to confirm transcription of the Wnt target gene AXIN2. Our results show that exogenous WNT3A induces β-catenin nuclear localization in ES cells and is associated with induction of TCF-mediated transcriptional activity and AXIN2 expression. Furthermore, Wnt/β-catenin activation is potentiated in the presence of Rspo2, and ES cells lines with the highest expression of LGRs show the strongest potentiation. Interestingly, however, exogenous WNT3A and Rspo2 did not induce Wnt activity in all cells. Together these data confirm that in the presence of exogenous ligand, Wnt/β-catenin signaling can be robustly activated in a subpopulation of ES cells and that signaling is potently enhanced in the presence of Rspo2 and LGRs. To our knowledge, this is the first evidence of R-spondins influencing Wnt activity in a non-epithelial cancer. These studies support the concept that R-spondin expression in the tumor microenvironment may contribute to ES pathogenesis by modulating Wnt/β-catenin signaling.
Citation Format: Elisabeth A. Pedersen, Christopher A. Scannell, Elizabeth R. Lawlor. R-spondin potentiates Wnt/β-catenin signaling in Ewing sarcoma cells. [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 5031. doi:10.1158/1538-7445.AM2013-5031
Abstract 5035: Stable transfection of the osteosarcoma cell line SaOs2, genetically transformed MSCs and osteoblasts with insulin-like growth factor 2 receptor.
Authors: Qu Huayi 
Published: August 2014

Abstract
Osteosarcoma (OS) is the most commonly seen primary malignant bone tumor in children and adolescents. With neo-adjuvant chemotherapy and other treatment regimens developed, event-free survival has increased to 70%, but the prognosis remains poor for those patients with unresectable or metastatic tumors or less responsive tumors. Insulin-like growth factor 2 (IGF2) and its receptor (IGF2R) are known to be over-expressed in several cancers including OS and have roles in regulating cellular growth and proliferation. Although IGF2R is thought to be a tumor suppressor, the research results in different experimental systems are variable.
Our recent studies have demonstrated that IGF2R is consistently over-expressed, relative to other known transmembrane receptors on the cell surface across, 4 standard and 10 patient-derived OS cell lines. To further elucidate the effects of IGF2R in OS, the OS cell line SaOS2 and two cell lines which were genetically transformed, as follows; mesenchymal stem cells (MSC) and osteoblasts (OB) with human telomerase reverse transcriptase and SV40Tag (MSC-TS and OB-TS) were transformed with IGF2R by lentiviral transfection. Drug resistant colonies were picked up 21 days after selection to obtain stably transfected SaOS2, MSC-TS, and OB-TS cell lines now named, SaI, MSC-TSI and OB-TSI, respectively. After cloning, the highest and the lowest IGF2R expression clones were selected from these different cell lines via real-time PCR and western blot for further analysis. The comparison between the lines and characterizations are underway. Initial results suggest introduction of IGF2R changes the phenotype by enhancing cellular motility. Further studies will be presented and include the following: 1) cell proliferation assays, 2) motility and migration assays, 3) subcutaneous tumorigenicity assays, 4) expression profiling, and 5) differentiation assays. In the context of osteosarcoma introduction of IGF2R enhances motility, which is unexpected given prior expectations based on the non-signaling nature of this receptor.
Citation Format: Huayi Qu. Stable transfection of the osteosarcoma cell line SaOs2, genetically transformed MSCs and osteoblasts with insulin-like growth factor 2 receptor. [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 5035. doi:10.1158/1538-7445.AM2013-5035
Abstract 5057: Activation of stroma-derived matrix metalloproteinases (MMPs) in a model of gastric cancer.
Authors: Kandola Sandhir , Holmberg Christopher , Ghesquiere Bart , Impens Francis , Gevaert Kris , Kumar J. Dinesh , Cash Nicole , Hegyi Peter , Vlatkovic Nikolina , Wang Timothy , Dockray Graham , Varro Andrea 
Published: August 2014

Abstract
Gastric cancer is the second commonest cause of cancer-related death worldwide. Within these tumors, it is increasingly recognised that signaling between myofibroblasts, an important stromal cell type, and cancer cells results in proliferation, invasion and metastasis. However, little is known of the range of proteins secreted by stromal cells ie their secretome, or the contribution of proteases in shaping the extracellular milieu ie the cancer degradome.
Using SILAC labeling and combined fractional diagonal chromatography (COFRADIC) we identified and quantified differential abundance and appearance of neo-N-termini (ie cleavage products) in the secretomes of gastric cancer-associated myofibroblasts (CAMs) compared with myofibroblasts from adjacent tissue (ATMs). There was decreased abundance of many proteins in CAM secretomes, but a small sub-set exhibited increased abundance including matrix metalloproteinases (MMP) -1 and -3. Cancer-restricted proteolytic cleavages included cleavage in the prodomains of MMP-1, -2 and -3 consistent with activation. Western blot, enzyme activity and cancer cell migration assays verified significantly increased abundance of active MMP-1, -2 and -3 in CAM media.
To establish the in vivo relevance of these findings we utilized a model in which primary gastric CAMs stimulate growth of MKN45 gastric cancer cells in xenografts in nude mice. MMP activity was imaged by fluorescence molecular tomography (FMT) employing MMPSense750 FAST™, which produces a fluorescent signal following MMP cleavage. CAMs enhanced xenograft growth by 6-fold (mean ± SEM, 29.6±21.1 vs 196.8±32.2 mm3 n=12, p=0.004). Using an experimental design that achieved similar volumes for tumors containing MKN45 cells alone and those containing MKN45+CAM, FMT assessment of tumor-related MMP activity in vivo indicated a two-fold increase in CAM-MKN45 xenografts compared with MKN45 cells alone (0.9±0.4 vs 1.8±0.8 fmol/mm3).
Stromal cell secretomes therefore contribute to the remodelling of the cancer cell microenvironment by activation of MMPs with consequences for cell migration and tumor growth. This system has potential for in vivo screening of anti-cancer therapies directed at the inhibition of stroma-stimulated tumor growth.
Citation Format: Sandhir Kandola, Christopher Holmberg, Bart Ghesquiere, Francis Impens, Kris Gevaert, J. Dinesh Kumar, Nicole Cash, Peter Hegyi, Nikolina Vlatkovic, Timothy Wang, Graham Dockray, Andrea Varro. Activation of stroma-derived matrix metalloproteinases (MMPs) in a model of gastric cancer. [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 5057. doi:10.1158/1538-7445.AM2013-5057
Abstract 5067: Human prostate epithelial cells secrete the tumor suppressor maspin as a soluble and exosome-associated protein .
Authors: Dean Ivory S. , Dzinic Sijana , Kaplun Alexander , Zou Yi , Mao Guangzhao , Sheng Shijie 
Published: August 2014

Abstract
Maspin, an epithelial-specific member of the serine protease inhibitor superfamily, inhibits tumor invasion and metastasis. Overall maspin expression is downregulated or lost in invasive and metastatic carcinomas. Maspin subcellular distribution among the nuclear, cytoplasmic and extracellular compartments is differentially regulated during tumor progression. Extracellular maspin has been shown to inhibit tumor cell invasion and motility in vitro. Also, recombinant maspin was shown to inhibit endothelial cell tube formation in vitro and tumor-angiogenesis in vivo. To date, the mechanisms governing maspin trafficking remain unclear. In the current study, we aim to investigate the mechanism and biological significance of maspin secretion. Normal and tumor cells known to secrete maspin, were treated with or without drugs targeting classical or non-classical secretory pathways. The CM were collected and analyzed for soluble and exosome-associated proteins. We also utilized atomic force microscopy to characterize the physical characteristics of exosomes. We found maspin as a soluble protein in the CM. Maspin secretion was not inhibited by the drug treatments. In fact, maspin secretion increased under drug treatment targeting the classical secretory pathway. Furthermore, maspin was detected in the exosomes. Under drug treatment, the overall increase in maspin secretion was accompanied by an increase in detection of maspin in the exosomes. Based on the drug treatment-induced changes in maspin's secretion patterns, we speculate that maspin is secreted via a non-classical pathway. In addition, we provide the first evidence that maspin is secreted inclusively as both a free and an exosome-encapsulated molecule in normal and tumor cells. The existence of extracellular maspin as a free and an exosome-associated molecule implies a dual role for maspin in the tumor microenvironment. These novel findings are currently under investigation since the tumor suppressive activity of exosome-associated maspin may be of particular significance in cell-cell communication, and tumor suppressive activity of free maspin may be associated with extracellular matrix remodeling.
Citation Format: Ivory S. Dean, Sijana Dzinic, Alexander Kaplun, Yi Zou, Guangzhao Mao, Shijie Sheng. Human prostate epithelial cells secrete the tumor suppressor maspin as a soluble and exosome-associated protein . [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 5067. doi:10.1158/1538-7445.AM2013-5067
Abstract 5088: Targeting the mTORC2/HIF-2alpha/p53 pathway in clear cell renal cell carcinoma.
Authors: Selvarajah Jogitha , Moumen Abdeladim , Carroll Veronica A. 
Published: August 2014

Abstract
Inactivating mutations in the von Hippel Lindau (VHL) tumour suppressor gene leads to both hereditary and sporadic clear cell renal cell carcinoma (ccRCC). Loss of VHL function results in stabilisation of hypoxia inducible factors 1 and 2 (HIF-1α and HIF-2α) with HIF-2α critical for renal tumour progression, angiogenesis and resistance to radiotherapy and chemotherapy. Previous work in VHL defective RCC cell lines that express HIF-2α has shown that loss of HIF-2α, either with siRNA or by reintroduction of VHL, increases phosphorylation and stabilisation of p53 which restored sensitivity to radiation and chemotherapy. Furthermore, as recent evidence suggests that mTORC2 is required for HIF-2α protein synthesis in VHL defective cells, we investigated the effects of pharmacological inhibition of HIF-2α with the mTORC1/2 kinase inhibitor pp242 alone or in combination with DNA damaging agents. Phosphorylated p53 and total p53 levels were significantly elevated in VHL competent RCC cells lacking HIF-2α as compared with VHL defective cells, confirming a role for HIF-2α mediated inhibition of p53. We analysed the effects of the mTORC1/2 kinase inhibitor pp242 on HIF-2α protein levels by Western blot. pp242 inhibited phosphorylation of both mTOR and its substrate p70S6K and significantly reduced protein levels of HIF-2α as compared with the mTORC1 inhibitor rapamycin, which had no effect on HIF-2α. Since the DNA damaging agent camptothecin (CPT) is a potent inhibitor of HIF-1α we also determined its effects on HIF-2α expression. CPT reduced HIF-2α protein levels in VHL defective RCC cells and increased p53 stabilisation, which was significantly augmented in VHL competent cells following complete loss of HIF-2α. Consistent with its inhibition of the p53 pathway, HIF-2α prevented both spontaneous and CPT induced apoptosis of RCC cells as demonstrated by FACS analysis of cells in sub G1 and Western blot analysis of cleaved PARP. Taken together these results confirm that HIF-2α negatively regulates p53, confers resistance to DNA damage induced apoptosis in VHL defective RCC cell lines and that further studies investigating the pharmacological inhibition of HIF-2α mediated by mTORC1/2 kinase inhibitors alone or in combination with DNA damaging agents for the treatment of ccRCC are warranted.
Citation Format: Jogitha Selvarajah, Abdeladim Moumen, Veronica A. Carroll. Targeting the mTORC2/HIF-2alpha/p53 pathway in clear cell renal cell 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 5088. doi:10.1158/1538-7445.AM2013-5088
Abstract 5101: Tanshinone II-A inhibits angiogenesis through down regulation of COX-2 in human colorectal cancer.
Authors: Li Qi 
Published: August 2014

Abstract
Tanshinone II-A Inhibits Angiogenesis through Down Regulation of COX-2 in Human Colorectal Cancer
Qi Li*, Li-Hong Zhou, Hua Sui, Yan Wang, Xuan Liu, Ning-Ning Liu, Pei-Hao Yin
Clinic Oncology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai China
*correspondence to Qi Li, Lzwf@hotmail.com
Angiogenesis plays a significant role in colorectal cancer (CRC). cyclooxygenase-2 (COX-2) is involved with multiple aspects of CRC angiogenesis. The aim was to investigate the inhibitory effects of Tan II-A(Tanshinone II-A, Tan II-A) on tumor growth in mice, as well as alteration of expression of COX-2 and VEGF in CRC. We established the mice xenograft model of C26 CRC cell line, and injected 0.5, 1, 2mg/kg of Tan II-A and 1mg/kg of 5-FU in different individuals respectively in vivo. Then, we assayed tumor weight and volume, evaluated microvascular density and expression of VEGF. COX-2 promoter and COX-2 plasmids were transfected into HCT-116 cells, followed by detection of COX-2 promoter activity by chemiluminescence, detection of COX-2 mRNA expression by fluorescence quantitative PCR. Taken together, the results showed Tan II-A could inhibit tumor growth and VEGF level in vivo. Cell experiment showed marked inhibitory effect of Tan II-A on COX-2 and VEGF of HCT-116 cell lines in a dose-dependent manner. That means Tan II-A can effectively inhibit tumor growth and angiogenesis of human colorectal cancer via inhibiting the expression level of COX-2 and VEGF.
Keywords: Tanshinone II-A - colorectal carcinoma - angiogenesis - cyclooxygenase 2 - vascular endothelial growth factor
Citation Format: Qi Li. Tanshinone II-A inhibits angiogenesis through down regulation of COX-2 in human colorectal cancer. [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 5101. doi:10.1158/1538-7445.AM2013-5101
Abstract 5109: Integrated micro nanotechnology for the sensitive isolation of circulating tumor cells.
Authors: Yoon Hyeun Joong , Kim Tae Hyun , Zhang Zhuo , Ramnath Nithya , Wicha Max S. , Hayes Daniel F. , Simeone Diane M. , Nagrath Sunitha 
Published: August 2014

Abstract
Introduction and Objective: Circulating tumor cells (CTCs) have been identified in the art in peripheral blood from cancer patients and are likely the origin of metastatic disease. Isolation and capture of CTCs represent a potential alternative to invasive biopsies during diagnosis of disease. In the recent past, vVarious microfluidic- based CTC isolation technologieschips have been developed to capture, indentify, sort, and enumerate CTCs. but, they need to be more sensitive, reliable, and specific for CTC detection in early stage cancer patients. Here we present a microfluidic CTC-chip with functionalized nanomaterials to capture and culture CTCs.
Methods: Micro-sized structuresposts functionalized with anti-epithelial-adhesion-molecule antibodies (Anti-EpCAM) are is the common approachstructures to capture CTCs. Here we present a microfluidic device with functionalized planar structure to capture CTCs. The CTC-chipmicrofluidic device has a PDMS chamber and patterned gold postatterns . functionalized with graphene oxides as nanomaterial and anti-epithelial-cell-adhesion-molecule antibodies (Anti-EpCAM). To investigate the capture efficiency before running cancer patients’ samples, a low number of cells were spiked into 1 mL of whole blood. We stained the captured cells with cytokeratin, CD45, and DAPI to identify CTCs. For clinical studies, we searched for CTCs in 10 pancreatic cancer patients, 6 breast cancer patients, 4 early lung cancer patients, and 5 healthy donors.
Results: It is much easier to scan, count and grow captured cells in comparison with previous micro-sized post-based structure. CTCs were isolated by assembling Graphene oxide on the planar structures for enhanced sensitivity. is easy to functionalize the surface and sensitive to detect biomaterial. The functionalized graphene oxide has been used to selectively capture MCF-7 breast cancer cells with a capture rate higher than 8790 %. This new CTC-chip identified CTCs in the peripheral blood of patients with pancreatic, breast, and lung cancer in 20 of 20 samples (100%).
Conclusion: We present a sensitive CTC-chip using graphene oxides as nanomaterial to isolate, capture, identify, and characterize extremely rare CTCs. This CTC-chipIt is much easier to scan, count, and grow captured cells in comparison with previous micro-sized post-based structure. We believe this CTC-chip offers great potential for clinical research on cancer.
Citation Format: Hyeun Joong Yoon, Tae Hyun Kim, Zhuo Zhang, Nithya Ramnath, Max S. Wicha, Daniel F. Hayes, Diane M. Simeone, Sunitha Nagrath. Integrated micro nanotechnology for the sensitive isolation of circulating tumor cells. [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 5109. doi:10.1158/1538-7445.AM2013-5109
Abstract 5124: 12-Lipoxygenase regulation of prostate cancer progression.
Authors: Tovar Elizabeth , Tucker Stephanie , Honn Kenneth V. 
Published: August 2014

Abstract
The underlying mechanism by which elevated omega-6 fatty acids, like arachidonic acid, lead to increased prostate cancer (PCa) risk is unclear. However, it is known that the 12-Lipoxygenase (12-LOX) enzyme that metabolizes arachidonic acid, as well as the 12(S)-HETE metabolic end product together with its cognate receptor, 12-HETER1, are strongly associated with PCa. 12(S)-HETE affects many cancer phenotypes including cell migration and metastasis. It causes the disassembly of complex cellular anchors to the substrata called hemidesmosomes that are comprised of numerous proteins, including ITGB4, which we identified as an interacting partner of 12-LOX. ITGB4 also physically interacts with c-Met at the cell membrane in multiple cell lines, in a cell line and stimulus-specific manner. PCa metastases have increased levels of both 12-LOX and c-Met. Therefore we sought to determine if ITGB4, c-MET and 12-LOX comprise a signaling axis. Our findings revealed that HGF-c-Met-induced invasion could be attenuated by enzymatic inhibitors of 12LOX, but that addition of exogenous 12(S)-HETE could not overcome the block. Furthermore, the physical interaction between β4 and c-Met remained unaffected by enzymatic inhibitors in PCa cells. Likewise, LC-MS analyses, showed that HGF stimulation did not lead to increased 12(S)-HETE production, and knockdown of 12-HETER1, by shRNA had no effect on HGF-induced invasion. Therefore, we hypothesize that 12LOX has dual functions whereby its enzymatic activity leads to hemidesmosome dissociation and cell release from the substrata, and its capacity to modulate c-MET/ITGB4 signaling may be attributed to a scaffolding function.
Citation Format: Elizabeth Tovar, Stephanie Tucker, Kenneth V. Honn. 12-Lipoxygenase regulation of prostate cancer 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 5124. doi:10.1158/1538-7445.AM2013-5124
Abstract 5130: Prostate circulating tumor cells metastasize to bone via E-selectin expressed on endothelial cells.
Authors: Gakhar Gunjan , YuLee Guang , Seandel Marco , Bander Neil , Nanus David 
Published: August 2014

Abstract
Abstract
Prostate cancer cells preferentially metastasize to bone. Studies show that the preferential adhesion is due to interactions between prostate tumor cells and E-selectin present on human bone marrow endothelial cells. In our study, we used a parallel flow chamber to identify if prostate circulating tumor cells (CTCs) metastasize using similar interactions. CTCs were investigated based on the fact that majority of prostate tumor cells exhibit prostate specific membrane antigen (PSMA) on their cell surface. Prostate cancer cells, MDA PCa 2b (MDA) were first labeled with anti-PSMA monoclonal antibody J591 conjugated with alexa fluor 488 (J591-488) that recognizes PSMA and is internalized following binding to PSMA. Anti-PSMA labeled MDA cells were then perfused over IL-β stimulated human umbilical vein endothelial cells (HUVECs) using parallel flow chamber under physiological blood flow. Both unlabeled and J591-488 labeled MDA cells showed similar rolling behavior on stimulated HUVECs. This suggests that prostate CTCs can be identified and their interactions with endothelial cells can be studied. Our study will potentially elucidate the mechanism involved in prostate cancer metastasis to bone.
Citation Format: Gunjan Gakhar, Guang YuLee, Marco Seandel, Neil Bander, David Nanus. Prostate circulating tumor cells metastasize to bone via E-selectin expressed on endothelial cells. [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 5130. doi:10.1158/1538-7445.AM2013-5130