bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4108: NKX6.1 suppresses cancer invasion and epithelial-mesenchymal transition in cervical cancer
Authors: Li Hsin-Jung , Yu Pei-Ning , Shih Yu-Lueng , Lin Ya-Wen 
Published: August 2015

Abstract
Cervical cancer is a common women malignancy worldwide. Recent reports emphasize inactivation of tumor suppressor genes are due to promoter hypermethylation in cervical cancer. Our previous study from methylation array has shown NK6 homeobox 1 (NKX6.1) is hypermethylated and downregulated in cervical cancer cell lines and clinical samples. Nkx6.1 contains a homeobox domain and functions as a transcription factor that regulates insulin-secreting β cell differentiation and neuronal fate determination. However, the biological function of NKX6.1 in cervical cancer remains elusive. Overexpression of NKX6.1 suppressed the transformation and invasive ability of HeLa and CaSki cells in vitro as well as decreased tumor formation and metastatic property in vivo. Conversely, knockdown of NKX6.1 strongly enhanced the transformation and invasiveness of SiHa cells. Epithelial-mesenchymal transition (EMT) has been shown to play a key role in the progression of uterine cervix carcinoma in situ (CIS) to invasive squamous cell carcinoma (SCC). NKX6.1 suppresses cancer invasiveness through coordinately increasing E-cadherin and decreasing Vimentin. Moreover, we demonstrated E-cadherin and vimentin were direct targets of NKX6.1 by luciferase reporter assay, electrophoretic mobility shift assay and chromatin immunoprecipitation quantitative polymerase chain reaction (Chip-qPCR) assay. Collectively, our data demonstrate NKX6.1 acts a tumor suppressor, and inhibits tumor invasion and metastasis by suppressing Epithelial-mesenchymal transition (EMT) in cervical cancer. These data imply that modulation of NKX6.1 expression may be a novel therapeutic option for treatment of cervical cancer.
Citation Format: Hsin-Jung Li, Pei-Ning Yu, Yu-Lueng Shih, Ya-Wen Lin. NKX6.1 suppresses cancer invasion and epithelial-mesenchymal transition in cervical cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4108. doi:10.1158/1538-7445.AM2015-4108
Abstract 4113: Characterization of a novel TIMP-2 deficient mouse model of experimental lung metastasis
Authors: Kumar Sarvesh 
Published: August 2015

Abstract
It is well known that the balance between matrix metalloproteinases (MMPs) and their endogenous inhibitors, the tissue inhibitors of metalloproteases (TIMPs) impacts tumor progression, thereby providing a novel avenue for cancer therapies. We have previously shown that forced TIMP-2 expression inhibits lung tumor growth independent of MMP inhibition via direct effects on tumor cells and modulation of the tumor microenvironment.
The current study was undertaken to examine how reduced TIMP-2 expression impacts lung tumor growth using a novel TIMP-2 deficient (T2D) mouse model in a clean C57BL6 background. This is first time we fully developed and characterized TIMP2 deficient mice, validated by genotyping, histologic staining, protein and mRNA expression. The deletion of TIMP2 gene down regulated the expression TIMP2 mRNA and protein as demonstrated by quantitative RT-PCR and immunoblot experiments, respectively. Further, we investigated the impact of TIMP2 deletion on Lewis lung carcinoma model of metastasis. Lewis lung carcinoma cells transfected with luciferase (LL/2-Luc-M38, Caliper) were intratracheally instilled in the lungs into group's have heterozygous (T2D+/−), homozygous (T2D−/−), and wild type (WT) littermates. Tumor progression was monitored by IVIS imaging after 2nd week of intratracheal instillation and continued till 4th week, mice were sacrificed and lungs were perfused and harvested for further analysis. The IVIS analysis showed higher tumor burden in TIMP2 deficient and heterozygous mice as compared to wild type littermate, suggested that TIMP2 deficiency aggravates tumor development in deficient mice. The effects of recombinant TIMP2 on tumor microenvironment and microarray experiments are currently in progress. The molecular mechanism underlying the observed activities will be investigated. Collectively, these preliminary findings suggest that TIMP2 may be a novel target for lung cancer therapy.
Citation Format: Sarvesh Kumar. Characterization of a novel TIMP-2 deficient mouse model of experimental lung metastasis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4113. doi:10.1158/1538-7445.AM2015-4113
Abstract 4120: Tasquinimod inhibits local invasion and metastases in two preclinical models of renal cell carcinoma
Authors: Shen Li , Pierron Valerie , Bauchet Anne-Laure , Nakhle Jessica , Orillion Ashley , Adelaiye Remi , Ciamporcero Eric , Meyer-Losic Florence , Eriksson Helena , Lassau Nathalie , Leguerney Ingrid , Schmidlin Fabien , Olsson Anders , Pili Roberto 
Published: August 2015

Abstract
Renal clear cell carcinoma (RCC) is the most common type of kidney cancer in adults and represents 80% of cases. At the time of diagnosis, approximately 30% of patients have distant metastases. In addition, one third of patients who have surgical resection of the primary tumor eventually relapse and develop metastases. Effective agents that help to prevent the development of metastatic RCC are not available. Tasquinimod is a novel agent in clinical development for metastatic castration-resistant prostate cancer that has been shown to have immunomodulatory, anti-angiogenic, and anti-metastatic properties. In the current study, we tested whether tasquinimod effectively inhibits local invasion and spontaneous lung metastases in RCC. Two preclinical mouse models of RCC were used for this purpose: a xenograft nude model A498 and a recently established patient-derived xenograft model, RP-R-02 LM. In both models, tasquinimod had a modest effect on the growth of the primary tumors implanted subcutaneously. Interestingly, in the A498 RCC model, gene expression analysis in tumors revealed that tasquinimod increased E-cadherin and decreased vimentin expression, suggesting an effect on reversal of epithelial-mesenchymal transition. In addition, tasquinimod significantly decreased CXCR4 expression, suggesting a reduction in invasiveness and metastatic phenotype. To further test this hypothesis, we used the RP-R-02 LM model that is able to develop lung metastases. Tasquinimod induced a 66% reduction of lung metastases in comparison with the control group (p&lt;0.01). Following tasquinimod treatment CD206+/Arginase 1+/high suppressive M2-polarized infiltrating macrophages were decreased in primary tumors, but even more so in the lung parenchyma and tumor nodules. In summary, our results demonstrate that tasquinimod has immunomodulatory and anti-metastatic properties in RCC models, providing a biological rationale for the clinical assessment of tasquinimod in RCC patients.
Citation Format: Li Shen, Valerie Pierron, Anne-Laure Bauchet, Jessica Nakhle, Ashley Orillion, Remi Adelaiye, Eric Ciamporcero, Florence Meyer-Losic, Helena Eriksson, Nathalie Lassau, Ingrid Leguerney, Fabien Schmidlin, Anders Olsson, Roberto Pili. Tasquinimod inhibits local invasion and metastases in two preclinical models of renal cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4120. doi:10.1158/1538-7445.AM2015-4120
Abstract 4126: CSF-1R inhibition blocks rhabdomyoscarcoma metastasis by polarizing macrophage differentiation
Authors: Evans Justin , Giles Amber , Reid Caitlin , Kaplan Rosandra 
Published: August 2015

Abstract
Colony Stimulating Factor 1 (CSF-1) is overexpressed by many tumors and predicts poor clinical outcome. CSF-1 expression results in the recruitment of CSF-1R (Fms)-expressing cells, which have been shown to suppress tumor immunity, thereby promoting cancer progression. PLX3397 is a novel, orally active small molecule inhibitor that selectively targets CSF-1R, KIT, and oncogenic FIT3. CSF-1R and KIT regulate the tumor, tumor microenvironment, and the pre-metastatic niche. However, it is unknown if targeting the myeloid CSF-1R expressing component of the tumor microenvironment regulates metastasis of pediatric tumors, such as rhabdomyosarcoma. Using the M3-9-M murine syngeneic orthotopic rhabdomyosarcoma model, we evaluated the effects of PLX3397 on primary tumor growth and metastatic progression. This highly metastatic rhabdomyosarcoma model has high infiltration of CSF-1R-expressing bone marrow-derived cells, which includes inflammatory monocytes, M2 macrophages, and myeloid-derived suppressor cells. Therefore, we hypothesized PLX3397 would have a direct effect on CSF-1R expressing M2 macrophages and inflammatory monocytes, resulting in decreased metastasis. Using both in vitro and in vivo models, we found that PLX3397 treatment decreased tumor-infiltrating M2 macrophages and skewed the differentiation of inflammatory monocytes toward M1 macrophages and development of immunocompetent mature dendritic cells. This resulted in impairment of spontaneous M3-9-M metastasis to secondary sites. In addition, we have found that the polarization of inflammatory monocytes reversed the T cell suppressive capability of tumor-derived myeloid cells. In conclusion, CSF-1R blockage with PLX3397 inhibited metastatic progression by decreasing the infiltration of M2 macrophages in the primary tumor and pre-metastatic niche. The polarization of inflammatory monocytes to dendritic cells restored the immune recognition and decreased metastasis. This data supports our rationale for testing PLX3397 in patients with high M2 infiltration and suggests value in this agent as an adjuvant setting for metastasis prevention in high-risk patients.
Note: This abstract was not presented at the meeting.
Citation Format: Justin Evans, Amber Giles, Caitlin Reid, Rosandra Kaplan. CSF-1R inhibition blocks rhabdomyoscarcoma metastasis by polarizing macrophage differentiation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4126. doi:10.1158/1538-7445.AM2015-4126
Abstract 4142: A role for apolipoprotein E in invasion in oral squamous cell carcinoma
Authors: Jayakar Sangeeta K. , Loudig Olivier D. , Brandwein-Gensler Margaret , Kim Ryung , Ow Thomas J. , Prystowsky Michael B. , Childs Geoffrey , Segall Jeffrey E. , Belbin Thomas J. 
Published: August 2015

Abstract
Oral squamous cell carcinomas (OSCC) have a poor patient prognosis, which is attributed to their invasive nature. Our goal was to identify a novel gene that is important in OSCC invasion. We utilized genome-wide expression data generated from OSCC patient tumor samples with varying degrees of invasiveness, as measured by the histological parameter worst pattern of invasion (WPOI). RNA extracted from tumors that were classified as either WPOI 5 or WPOI 3, two categories of tumors that have been previously shown to correlate with local recurrence and poor overall survival, were evaluated utilizing two microarray platforms. When both gene expression datasets were analyzed, we identified 104 genes that were overexpressed at least 1.5-fold in the WPOI type 5 tumors compared to the less invasive WPOI type 3 tumors. Fifty genes were chosen based on functional criteria to be assessed using in vitro invasion assays following treatment with siRNA pools. In our initial screen, 16 of these genes showed significant reductions in in vitro invasion when their expression was knocked down by siRNA treatment in the UMSCC1 cell line. The gene with the greatest effect on invasion from the screen was Apolipoprotein E (APOE).
In addition to in vitro invasion, we also observed that matrix degradation and mature invadopodia numbers were decreased with APOE knockdown in the UMSCC1 cell line, suggesting that APOE affects invasion in part through regulating invadopodial activity. We observed that APOE knockdown resulted in decreased phospho-cJun and phospho-ERK1/2 in the UMSCC1 and PCI-15B cell lines. Since cJun is a component of the AP1 transcription factor, we assessed AP1 promoter activity using a luciferase reporter assay. We found that AP1 promoter activity decreased with APOE knockdown in both cell lines. Our proposed model is that APOE knockdown suppresses the cells’ ability to invade via decreased activation of ERK1/2 and cJun, thus reducing AP1 transcriptional activity. Further studies are being done to determine which downstream targets of AP1 are being altered with APOE knockdown and if those targets are known to be implicated in invasion.
Citation Format: Sangeeta K. Jayakar, Olivier D. Loudig, Margaret Brandwein-Gensler, Ryung Kim, Thomas J. Ow, Michael B. Prystowsky, Geoffrey Childs, Jeffrey E. Segall, Thomas J. Belbin. A role for apolipoprotein E in invasion in oral squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4142. doi:10.1158/1538-7445.AM2015-4142
Abstract 4149: Epidermal growth factor-induced cyclooxygenase-2 enhances head and neck squamous cell carcinoma metastasis through fibronectin up-regulation
Authors: Hsu Jinn-Yuan , Chang Kwang-Yu , Chen Shang-Hung , Lee Chung-Ta , Chang Sheng-Tsung , Cheng Hung-Chi , Chang Wen-Chang , Chen Ben-Kuen 
Published: August 2015

Abstract
Epidermal growth factor receptor (EGFR) activation is a major cause of cell metastasis in many cancers, such as head and neck squamous cell carcinoma (HNSCC). However, whether the induction of cyclooxygenase-2 (COX-2) correlates with EGF-enhanced HNSCC metastasis remains unclear. Interestingly, we found that EGF induced COX-2 expression mainly in HNSCC. The tumor cell transformation induced by EGF was repressed with COX-2 knockdown, and this repression was reversed by simultaneously treating the cells with EGF and prostaglandin E2 (PGE2). The down-regulation of COX-2 expression or inhibition of COX-2 activity significantly blocked EGF-enhanced cell migration and invasion, but the addition of PGE2 compensated for this blockage in COX-2-knockdown cells. Interestingly, COX-2 depletion inhibited EGF-induced matrix metalloproteinase-1 (MMP-1), MMP3 and fibronectin expression and Rac1/cdc42 activation; this reduction in MMPs and the fibronectin/Rac1/cdc42 axis by the depletion of COX-2 was also rescued when the cells were treated with PGE2. Furthermore, the depletion of fibronectin impeded the COX-2-enhanced binding of HNSCC cells to endothelial cells and tumor cells metastatic seeding of the lungs. These results provide new insight that EGF-induced COX-2 enhances HNSCC metastasis via activation of the fibronectin signaling pathway. The inhibition of COX-2 expression and activation suggests a potential strategy for the treatment of EGFR-mediated HNSCC metastasis.
Citation Format: Jinn-Yuan Hsu, Kwang-Yu Chang, Shang-Hung Chen, Chung-Ta Lee, Sheng-Tsung Chang, Hung-Chi Cheng, Wen-Chang Chang, Ben-Kuen Chen. Epidermal growth factor-induced cyclooxygenase-2 enhances head and neck squamous cell carcinoma metastasis through fibronectin up-regulation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4149. doi:10.1158/1538-7445.AM2015-4149
Abstract 415: Differential expression of therapeutic targets across tumor micro-environments and at infiltrative margins in glioblastoma
Authors: McCrary Myles R. , Gutman David , Dunn William , Kaluzova Milota , Bouros Alexandros , Williams Merete , Zhang Xialong , Cooper Lee AD , Hadjipanayis Constantinos G. , Brat Daniel J. 
Published: August 2015

Abstract
Glioblastoma (GBM; WHO grade IV) is the most common primary brain tumor and is associated with a poor prognosis. The Cancer Genome Atlas (TCGA) project has revealed discrete molecular classes of GBM, raising the possibility of targeted therapies. Molecular heterogeneity across tumor microenvironments presents a challenge to targeted therapy, since effective therapies will be directed at targets near resection margins rather than those expressed in bulk resected tumor. To this end, we initiated studies to delineate spatial and temporal variations of potential druggable targets in GBM, focusing on potential protein targets at the tumor margin.
By utilizing 5-ALA (5-aminolevulinic acid, an oral compound that accumulates in malignant glioma cells and is metabolized to the fluorescent agent, protoporphyrin IX) fluorescence-guided surgery of primary and recurrent GBMs in a Phase II clinical trial, we were able to precisely define perinecrotic, bulk, and marginal regions within the glioma intraoperatively in order to sample tissues for further molecular characterization. Tissues harvested from these 3 tumor regions from 5 patients were analyzed using reverse phase protein array (RPPA) analysis with 218 antibodies (Array-Pro). Relative protein levels were determined and a heatmap was generated as a hierarchical cluster using Pearson Correlation and a center metric.
Forty proteins had statistically significant altered expression at the tumor margin compared with bulk and perinecrotic tumor. Among these, STAT1, STAT2, STAT3, PDGFRA, PDGFRB, EGFR, JAK2, MET, PIK3R1, and ERBB2 were upregulated at the margins and are potential druggable targets worthy of further investigation. Also upregulated at the tumor margins were NFKBIA, JUN, ZAP70, GAPDH, and CD28, whereas STAT5A, EFNB2, NTRK1, KDR, FGFR1, PTPRA, IFNGR1, and EFNB1 were downregulated at margins.
Our data demonstrate significant variation of druggable targets and signaling networks across tumor microenvironments in GBM. Marginal tumor differs from bulk and perinecrotic tumor by the relative overexpression of STAT proteins, growth factor receptors, and receptor kinases. Directing therapy at networks and targets present at the tumor margin of GBM may be an effective strategy.
Citation Format: Myles R. McCrary, David Gutman, William Dunn, Milota Kaluzova, Alexandros Bouros, Merete Williams, Xialong Zhang, Lee AD Cooper, Constantinos G. Hadjipanayis, Daniel J. Brat. Differential expression of therapeutic targets across tumor micro-environments and at infiltrative margins in glioblastoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 415. doi:10.1158/1538-7445.AM2015-415
Abstract 4151: Interleukin 17 promotes tumor progression in oral squamous cell carcinoma
Authors: Avadhani Avadhoot , Milne Trudy , Seymour Gregory , Rich Alison 
Published: August 2015

Abstract
Background: Interleukin 17 (IL17) is a proinflammatory cytokine with increased expression in some cancers. It has been demonstrated to exhibit both pro- and anti-tumor effects. Our group has previously demonstrated expression of IL17 from multiple cells in oral squamous cell carcinoma (OSCC) such that the aim of the current study was to examine the role of IL17 in OSCC progression.
Methods: The cytoplasmic SEFIR sequence of the transmembrane IL17 receptor (IL17R) was detected in formalin Fixed Paraffin Embedded (FFPE) OSCC tissues (n = 14) using immunohistochemistry. Soluble IL17R was detected in the cell culture supernatants of three OSCC cell lines (SCC4, SCC15 and SCC25) using a sandwich ELISA. Human recombinant IL17 over a range of concentrations (0, 10, 50, 100 ng/mL) was used to stimulate the three OSCC cell lines.Proliferation at 0, 24, 48, 72 hours was then assayed by cell titer blue and invasion at 48 hours using a QCM ECMatrix cell invasion assay. The data were analyzed using unpaired Student's t test and ANOVA as appropriate using GraphPad Prism 6 software.
Results:Cytoplasmic expression of transmembrane IL17R was detected in all FFPE OSCC tissues. Soluble IL17R was detected in the cell culture supernatants of all three OSCC cell lines and its concentration increased in a time dependent manner. SCC25 had significantly higher concentration of soluble IL17R at 72 hours than SCC4 and SCC15. The rate of proliferation of all three OSCC cell lines was not affected with the addition of recombinant IL17. However, IL17 promoted the in vitro invasion of SCC25 and SCC15 in a dose dependent manner. At the highest concentration of IL17 (100ng/mL), the SCC25 cells showed significant invasion compared to control cells (p&lt;0.05), while the SCC15 cells exhibited significant invasion at both 50ng/mL and 100 ng/mL of IL17 (p&lt;0.05). In contrast, IL17 failed to stimulate invasion in the SCC4 cell line.
Conclusions: This study is the first to demonstrate in vitro association between IL17 and possible invasion of OSCC. The exact mechanism by which IL17 enhances invasion and thereby tumor progression in vivo remains to be elucidated and could involve a number of different mechanisms including cell adhesion and extracellular matrix proteins.
Citation Format: Avadhoot Avadhani, Trudy Milne, Gregory Seymour, Alison Rich. Interleukin 17 promotes tumor progression in oral squamous cell carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4151. doi:10.1158/1538-7445.AM2015-4151