bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3156: Autocrine signaling through release of ATP and activation of P2X7 receptor promotes migration of human lung cancer cells
Authors: Takai Erina , Tsukimoto Mitsutoshi , Harada Hitoshi , Hiasa Miki , Moriyama Yoshinori , Kojima Shuji 
Published: May 2015

Abstract
Cell migration is essential step of cancer metastasis. TGF-β1 plays a key role in cancer progression through induction of various biological effects, including cell migration. Extracellular nucleotides, such as ATP, released from cells play a role in signaling through activation of P2 receptors. However, it remains poorly understood whether ATP-P2 receptor signaling is involved in cancer progression. We show here that release of ATP followed by activation of P2X7 receptors plays a key role in migration of lung cancer cells. Treatment with TGF-β1 facilitated migration of human lung cancer A549 cells, which was blocked by pretreatment with ecto-nucleotidase and P2 receptor antagonists in a wound healing-based assay and Transwell assay. ATP and P2 agonists facilitated cell migration. TGF-β1-induced actin remodeling, which contributes to cell migration, was also suppressed by pretreatment with ecto-nucleotidase and P2 receptor antagonists. Knockdown of P2X7 receptor suppressed TGF-β1-induced migration and actin remodeling. These results indicate the involvement of TGF-β1-induced ATP release in cancer cell migration, at least in part, through activation of P2X7 receptors. We found that TGF-β1 caused release of ATP from A549 cells within 10 minutes. Both ATP-enriched vesicles and expression of a vesicular nucleotide transporter (VNUT) SLC17A9, which is responsible for exocytosis of ATP, were found in cytosol of A549 cells. TGF-β1 failed to induce release of ATP from SLC17A9-knockdown cells. TGF-β1-induced cell migration and actin remodeling were also decreased in SLC17A9-knockdown cells. These results suggest the importance of exocytosis of ATP in cell migration. In addition, treatment with specific antagonist of P2X7 receptor (A438079) suppressed TGF-β1-induced migration of another human lung cancer H292 cells, but not of non-cancerous BEAS-2B cells. H292 cells released ATP in response to TGF-β1 stimulation, while BEAS-2B cells did not. Furthermore, in high-motile human lung cancer PC-9 cells, a large amount of ATP was released under standard cell culture condition, and P2X7 receptor was constitutively activated. Blockade of P2X7 receptor significantly suppressed motile activity of PC-9 cells. Taken together, these results suggest that autocrine signaling through release of ATP and activation of P2X7 receptor play a role in migration of human lung cancer cells.
Citation Format: Erina Takai, Mitsutoshi Tsukimoto, Hitoshi Harada, Miki Hiasa, Yoshinori Moriyama, Shuji Kojima. Autocrine signaling through release of ATP and activation of P2X7 receptor promotes migration of human lung cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3156. doi:10.1158/1538-7445.AM2014-3156
Abstract 3157: An Rgnef (p190RhoGEF/Arhgef28) signaling axis regulates ovarian cancer progression
Authors: Miller Nichol L. G. , Tancioni Isabelle , Uryu Sean , Kleinschmidt Elizabeth G. , Connolly Denise C. , Schlaepfer David D. 
Published: May 2015

Abstract
Ovarian cancer is a leading cause of cancer death in women, is usually diagnosed in late stage, and currently has insufficient effective targeted therapies. The primary mode of metastasis is unique in that dissemination occurs via tumor cell shedding into the peritoneal cavity, survival and growth within ascites, and re-adhesion and proliferation of tumor cells at sites within the abdomen. Therefore, molecules and pathways involving cell adhesion and survival in non-adherent environments are of particular interest. Rgnef (p190RhoGEF/Arhgef28) is a Rho family guanine exchange factor that associates with focal adhesion kinase (FAK) and functions both as a downstream target of FAK tyrosine kinase activity and a regulator of FAK activity following integrin stimulation. FAK regulates cell adhesion, migration, and survival. Expression of FAK is associated with poor clinical outcome, its activity (as measured by FAK Y397 phosphorylation) is frequently increased in serous ovarian carcinomas, and it is currently under investigation as a therapeutic target in ongoing clinical trials. Immunohistochemical analysis of human tumor tissue arrays with antibodies specific for Rgnef and FAK pY397 reveal a positive correlation between Rgnef expression and FAK activation and increased stage/grade of serous-type ovarian cancer. Stable knockdown of Rgnef in human ovarian carcinoma cells grown as subcutaneous or intraperitoneal xenografts in mice suggest that Rgnef may play roles in both primary tumor growth and ascites-associated cell survival and spread. Here, we use a transgenic model of spontaneous ovarian cancer (MISIIR-T-Antigen) to test the role of Rgnef in ovarian cancer progression. Female Rgnef+/+;TAg+ and Rgnef-/-;TAg+ mice were monitored by ultrasound imaging from age 12 to 17 weeks before euthanasia. Rgnef-/-;TAg+ tumors were significantly smaller (p=0.0006) and contained fewer Ki67 positive cells than Rgnef+/+;TAg+ controls at 17 weeks. Tumors and ovarian carcinoma cells isolated from Rgnef-/-;TAg+ mouse ascites have reduced FAK pY397 and increased E-cadherin expression compared to controls, suggesting that loss of Rgnef results in a less aggressive tumor phenotype. These findings identify Rgnef as a component in signaling pathways promoting FAK activation and regulating ovarian tumor progression.
Citation Format: Nichol L. G. Miller, Isabelle Tancioni, Sean Uryu, Elizabeth G. Kleinschmidt, Denise C. Connolly, David D. Schlaepfer. An Rgnef (p190RhoGEF/Arhgef28) signaling axis regulates ovarian cancer progression. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3157. doi:10.1158/1538-7445.AM2014-3157
Abstract 3158: Identifying novel genes critical to invasion in head and neck squamous cell carcinoma
Authors: Jayakar Sangeeta K. , Loudig Olivier D. , Brandwein-Gensler Margaret , Kim Ryung S. , Prystowsky Michael B. , Segall Jeffrey E. , Belbin Thomas J. 
Published: May 2015

Abstract
Oral squamous cell carcinomas (OSCC) have a poor patient prognosis, which is attributed to their invasive nature. Our goal was to identify specific genes that are important in OSCC invasion utilizing genome-wide expression data generated from HNSCC patient tumor samples of varying degrees of invasiveness. Invasion was assessed using the histological parameter pattern of invasion (POI), which describes the manner in which tumor cells infiltrate the normal tissue at the tumor-host interface. POI was used to categorize tumor resections into five types (1-5), type 5 being the most invasive with a “widely dispersed pattern of tumor infiltrate” and correlated significantly with the appearance of local recurrence and decreased overall survival of the patients after surgery. Global gene expression data was acquired from analysis of the surgical resections from 9 OSCC patient samples collected at Montefiore Medical Center between 2001 and 2008. Tumor cell-enriched cores from FFPE samples were used to generate global gene expression data using the Illumina® WG-DASL array to compare gene expression in four tumors that were WPOI type 5 and five tumors that were WPOI type 3. RNA extracted from flash-frozen tumor samples following surgical resection from the same patients was used to obtain global gene expression data using the Illumina® HumanHT-12 v4 Expression BeadChip. Candidate genes were assessed using in vitro invasion assays following siRNA knockdown of specific gene expression using siGENOME smart pools from Dharmacon. 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 initially tested based on functional criteria. In our initial screen, 16 of these genes showed a significant reduction in tumor cell invasion when expression was knocked down by siRNA in an OSCC cell line. The top gene from the screen, APOE, is now being further examined to determine a mechanistic explanation for its effect on invasion. Downstream signaling of APOE receptors and how this may interact with invasion are currently being assessed. This initial screen of global gene expression data in combination with pattern of invasion has revealed APOE as a novel gene that may play a critical role in OSCC invasion.
Citation Format: Sangeeta K. Jayakar, Olivier D. Loudig, Margaret Brandwein-Gensler, Ryung S. Kim, Michael B. Prystowsky, Jeffrey E. Segall, Thomas J. Belbin. Identifying novel genes critical to invasion in head and neck squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3158. doi:10.1158/1538-7445.AM2014-3158
Abstract 3159: ZAR2 transcriptionally represses the ATPase ATP6V0A4 to negatively regulate invasiveness of breast cancer cells
Authors: Misra Smita , Chaudhuri Gautam 
Published: May 2015

Abstract
ZAR2 is a recently characterized RNA-dependent transcriptional repressor protein that is implicated in the cell cycle-dependent regulation of BRCA2 gene expression. We have shown previously that ZAR2 mRNA is transcribed from the overlapping bi-directional promoter of the BRCA2 gene, binds to the overlapping promoter and prevents the expression of BRCA2 through chromatin remodeling. ZAR2 level is significantly low in the invasive breast cancer cells and tissues as compared to the non-invasive cells. Knockdown of ZAR2 in the non-invasive breast cancer cells increased the in vitro invasiveness of these cells whereas forced expression of ZAR2 in the invasive breast cancer cells prevented their invasiveness. To understand the possible mechanism of ZAR2-mediated regulation of invasiveness of the breast cancer cells we studied differential gene expression in the ZAR2 knocked down non-invasive breast cancer cells by RNA-seq analysis. One of the genes that are significantly elevated in the ZAR2 knocked down cells is the invasion determining enzyme ATP6V0A4. ATP6V0A4 is transcribed from an overlapping bi-directional promoter along with its partner TMEM213 in the breast cancer cells. We found that both ATP6V0A4 and TMEM213 levels are increased in the ZAR2 knocked down cells whereas cells with forced expression of ZAR2 have a significant decrease in the levels of these proteins. ZAR2 cell cycle dependently binds to the ATP6V0A4/TMEM213 gene promoter to repress the activity of this bi-directional promoter. This study thus reports a new pathway for the regulation of the invasiveness of breast cancer cells. Supported in part by DOD grants BC990678 and BC050641 to GC and NIH grant 1U54RR026140 to SM.
Citation Format: Smita Misra, Gautam Chaudhuri. ZAR2 transcriptionally represses the ATPase ATP6V0A4 to negatively regulate invasiveness of breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3159. doi:10.1158/1538-7445.AM2014-3159
Abstract 3167: Myosin-Va involvement in malignant properties of melanoma cells
Authors: Ramao Anelisa , Pontes Carmen Lucia S. , Espreafico Enilza M. 
Published: May 2015

Abstract
Evidence has emerged on the role of myosin-Va in cancer progression. Recently a role for myosin-Va in tumor cell migration, invasion and metastasis has been described for cancer cell lines from lung, colon, breast and prostate. One of the objectives of this work was to assess a possible role to myosin-Va in melanoma cells. We knocked down the expression of endogenous myosin-Va by using a lentiviral vector expressing a shRNA (shMYO5A) designed to target MYO5A mRNA in 1205Lu - a human melanoma cell line and functional studies were conductedonce efficient knockdown was attained. Myosin-Va depletion was showed to have no effect in cell proliferation as determined by crystal violet staining at a time course interval. However, cell viability was significantly reduced in MYO5A depleted cells compared to control ones. Also, a higher apoptotic index was observed in those cells suggesting that myosin-Va is involved in tumor growth. Subsequentlywe performed a cell migrationanalysis and found rates 60% lower for MYO5A depleted cells. This effect could have been caused by impairment in cell adhesion to the substrate as we could verify by a dynamic adhesion assay in different types of matrix. Adhesion ability to collagen, fibronectin, lamminin and matrigel was reduced in MYO5A depleted cells. Also, cell surface levels of integrins were lower for shMYO5A transfected cells compared to control as measured by flow cytometry approach and imply a defect in cell adhesion and migration caused by MYO5A knock-down.Next we evaluate cell survival under detachment conditions by plating cells on non-adherent surfaces and measuring cell death after 24h by annexin-V staining at cell surface. Results suggested that myosin-Va protects cells from anoikis (apoptosis caused by detachment) since MYO5A depleted cells presented higher rates of apoptosis compared to wild-type. Colony formation in soft agar showed that myosin-Va is required for anchorage-independent growth since depleted cells presented fewer and smaller colonies 30 days after culturing under those conditions. These findings indicate that myosin-Va is involved in several steps of tumor growth and progression including cell survival after detachment of the initial tumor mass and colonization of new microenvironments, suggesting that its upregulation in melanoma cells contributes to the rise of malignancy in this type of cancer.
Citation Format: Anelisa Ramao, Carmen Lucia S. Pontes, Enilza M. Espreafico. Myosin-Va involvement in malignant properties of melanoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3167. doi:10.1158/1538-7445.AM2014-3167
Abstract 3172: Detection of viral HPV DNA in sporadic colorectal cancers in relation to CpG island methylator phenotype (CIMP)
Authors: Bernabe-Dones Raul D. , Villavicencio-Torres Wesley , Munoz-Masso Cristina , Reyes -Medina Yaritza , Marcano-Bonilla Lorena , Perez-Cantalapiedra Hector , Fonseca-Williams Sharon , Lacourt-Ventura Mercedes Y. , Yamamura Yasuhiro , Rodriguez Nayra , Cruz-Correa Marcia R. 
Published: May 2015

Abstract
Background: There is evidence that insertion of viral DNA into a mammalian genome can lead to alterations of methylation patterns. To test the possible correlation between epigenetic modulations in colorectal cancer (CRC) and HPV infection, we conducted a MSP- PCR methylation analysis. The aim of the present study was to examine the presence of DNA sequences of human papillomavirus (HPV) in CRC and correlate this data with their CIMP status.
Methods: Thirty-seven colorectal adenocarcinomas were screened for HPV infection using nested-PCR (PGMY09/11-GP5+/6+). HPV typing was performed by type-specific PCR for HPV 16 and HPV 18 DNA. Eight CpG island methylator phenotype (CIMP) genes specific to sporadic colorectal cancer (MLH1, CACNA1G, NEUROG1, IGF2, SOCS1, RUNX3, CDKN2A and CRABP1) were assessed by methylation-specific PCR.
Results: Thirty-seven CRC cases (mean age at diagnosis 63.2 ± 11.7 years; 17 males) were evaluated. Tumors were mostly located in the left colon (73.0%), and moderate differentiated (61.1%). HPV's DNA was identified in 23 of 37 (62.2%) CRC studied cases. CRABP1 (59.5%), CDKN2A (37.8%), MLH1 (13.5%), and NEUROG1 (10.8%) were the most commonly methylated genes with a discrete clinic-pathological phenotype and gene methylation pattern. No statistical associations between HPV status and methylation patterns were found, after correlation with tumor staging, cell differentiation, tumor location, and life style.
Discussion: We reported that the CIMP-hypermethylation may be less frequent among Puerto Ricans Hispanics than previously reported in other racial and/or ethnic groups. However, a correlation could not be established (p&gt;0.5) between HPV infection and the aberrant methylation pattern in the genes panel. We observed the trend that HPV is more prevalent in CRC without aberrant methylation patterns. This might suggest that hypermethylation could be a protective factor for HPV infection. This inference may mean that HPV positive colorectal cancer could represent a distinctive clinical and epidemiological condition compared with HPV-negative CRC.
Citation Format: Raul D. Bernabe-Dones, Wesley Villavicencio-Torres, Cristina Munoz-Masso, Yaritza Reyes -Medina, Lorena Marcano-Bonilla, Hector Perez-Cantalapiedra, Sharon Fonseca-Williams, Mercedes Y. Lacourt-Ventura, Yasuhiro Yamamura, Nayra Rodriguez, Marcia R. Cruz-Correa. Detection of viral HPV DNA in sporadic colorectal cancers in relation to CpG island methylator phenotype (CIMP). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3172. doi:10.1158/1538-7445.AM2014-3172
Abstract 3174: The host genomic environment of the provirus determines the abundance of BLV-infected clones
Authors: Gillet Nicolas , Brogniez Alix de , Renotte Nathalie , Gutierrez Geronimo , Alvarez Irene , Trono Karina , Willems Luc 
Published: May 2015

Abstract
Human T-lymphotropic Virus 1 (HTLV-1) induces a persistent infection that remains generally asymptomatic. Nevertheless, in a small proportion of individuals and after a long latency, HTLV-1 infection leads to leukemia or lymphoma. Onset of clinical manifestations correlates with a persistently elevated number of infected cells. Because the vast majority of cells are infected at early stages, primary infection is a crucial period for HTLV-1 persistence and pathogenesis. Since HTLV-1 is transmitted through breast feeding and because systematic population screenings are rare, there is a lack of available samples at early infection. Therefore, we addressed this question in a closely related animal model by inoculating cows with Bovine Leukemia Virus (BLV). Initial BLV provirus insertion is not random but occurs nearby transcribed genomic regions (genes and CpG islands). This pattern of integration is highly similar to the observed pattern in HTLV-1 in vitro experiments. Later on, the proportion of clone carrying a provirus in transcribed regions decreases to reach levels comparable to those observed in asymptomatic HTLV-1 carriers. The negative selection against BLV clones carrying a provirus nearby transcribed regions is more efficient in animals that will subsequently present low proviral load set point. Finally, like for HTLV-1 infected people, the abundance of the BLV infected clones correlates with the integration of the provirus in transcribed regions. These observations highlight the high similarities between BLV and HTLV-1 clonality and suggest a key role exerted by the host immune system during primary infection.
Citation Format: Nicolas Gillet, Alix de Brogniez, Nathalie Renotte, Geronimo Gutierrez, Irene Alvarez, Karina Trono, Luc Willems. The host genomic environment of the provirus determines the abundance of BLV-infected clones. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3174. doi:10.1158/1538-7445.AM2014-3174
Abstract 3175: The splice variant of the human papillomavirus 16 E6 protein, E6*, displays anti-tumor properties in vivo
Authors: Evans Whitney , Filippova Maria , Aragon Robert , Filippov Valeri , Reeves Mark , Duerksen-Hughes Penelope 
Published: May 2015

Abstract
High-risk types of the human papillomavirus (HPV) are the causative agents of nearly all cases of cervical cancer, as well as a significant number of head, neck, penile, vulvar, and anal cancers. Like many other viruses with small genomes, HPV (∼8 kb) utilizes numerous mechanisms to increase the capacity of its genome to encode the proteins necessary for successful completion of its infectious life cycle, including alternative splicing. Studies over the past few decades have focused intensively on the activities and roles of E6 proteins from high-risk types of HPV during the process of cellular transformation, clearly implicating E6 as a major transforming agent. In contrast, the role of the smaller splice isoform, E6*, in the carcinogenic process (if any) has not yet been established. In the present study, we examined the behavior of the E6* protein during tumor growth in an in vivo nude mouse xenograft model. We created E6*-expressing SiHa (HPV+) and C33A (HPV-) cells, then examined the ability of both the parental and E6*-expressing cells to form tumors in nude mice. The difference in tumor size observed in the presence and absence of E6*, when expressed in both HPV+ (SiHa) and HPV- (C33A) cells, was dramatic and consistent, strongly indicating an anti-oncogenic role for E6* in this context. Interestingly, we found that tumor growth inhibition by E6* was greater in tumors derived from SiHa cells, which are HPV16-positive, than in tumors produced by C33A, which are HPV-negative. This difference implies that E6* acts by interfering with the oncogenic activity of the full-length protein as well as through one or more HPV-independent mechanisms. Consistent with this idea, we found that E6* does indeed bind to the full-sized isoform and inhibits its ability to accelerate degradation of p53 and procaspase 8. These data represent the first demonstration of biologically-relevant E6* activities distinct from those of the full-length isoform in vivo. The significance of these findings in the context of human cancers is in the possibility of mimicking or replicating the anti-oncogenic activity of E6* in such a way as to provide therapeutic benefit.
Citation Format: Whitney Evans, Maria Filippova, Robert Aragon, Valeri Filippov, Mark Reeves, Penelope Duerksen-Hughes. The splice variant of the human papillomavirus 16 E6 protein, E6*, displays anti-tumor properties in vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3175. doi:10.1158/1538-7445.AM2014-3175