bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1513: Mesenchymal stromal cells enhance the malignant potential of human colorectal cancer cells by inducing epithelial to mesenchymal transition-related phenomena
Authors: Mele Valentina , Muraro Manuele Giuseppe , Calabrese Diego , Giovenzana Chiara , Heberer Michael , Bocelli-Tyndall Chiara , Terracciano Luigi , Spagnoli Giulio Cesare , Iezzi Giandomenica
Published: June 2012

Background: Mesenchymal stromal cells (MSCs) are recruited to primary and metastatic sites of several tumour types, including colorectal cancer (CRC), and might contribute to tumour progression. The actual role played by MSCs and the mechanisms underlying MSC-tumour interactions remain to be clarified. We investigated the effects of human bone-marrow-derived MSCs (BM-MSCs) on CRC, in vitro and in vivo.

Material and methods: Human established CRC cell lines were cultured in the presence or absence of BM-MSCs, in direct contact or in transwell plates. After a five day culture, tumour cell proliferation was assessed by differential cell counts, surface molecule expression was analyzed by flow cytometry, and production of soluble factors in culture supernatants was measured by Raybio antibody array® and ELISA. Tumour cells, sorted upon co-culture by flow cytometry, were evaluated for the expression of epithelial to mesenchymal transition (EMT)-related genes by quantitative PCR and for in vitro invasiveness, by chemoinvasion assay. Furthermore, their tumorigenicity was assessed upon injection in NOD/SCID mice and developing tumours were analyzed by immunofluorescence.

Results: MSCs significantly increased tumour cell proliferation and decreased CD44 expression, independently of cell-to-cell contact. Analysis of co-culture supernatants revealed higher amounts of IL-6, MCP-1, RANTES and Angiogenin, in comparison to supernatants derived from single cultures. Moreover increased expression of several EMT-related genes, including SNAI2, TWIST, N-Cadherin, was detected on CRC cells sorted upon co-culture as compared with controls. Importantly, CRC cells co-cultured with MSCs showed higher invasive behaviour in vitro, than CRC cells cultured alone. No significant changes were observed in tumorigenicity. However, tumours originated from tumour cells co-cultured with MSCs showed a significantly higher vessel density as compared to controls.

Conclusions: MSCs reduce adhesiveness, induce expression of EMT-related genes and increase proliferation, invasiveness and angiogenic potential of CRC cells. These effects might contribute to CRC progression and spreading.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1513. doi:10.1158/1538-7445.AM2011-1513
Abstract 1522: Spheroid formation causes a decrease in ovarian cancer cell growth rate that persists through dissociation and growth as a monolayer
Authors: Pease Jillian C. , Tirnauer Jennifer S.
Published: June 2012

Ovarian cancer has the highest mortality of all cancers of the female reproductive system. It accounts for three percent of all cancers in women and is the fifth leading cause of cancer-related death among women in the United States. Ovarian cancer cells can metastasize to the uterus, fallopian tubes, and pelvis tissues as well as to other organs. Even with optimal treatment there is a high probability that the cancer will recur, suggesting that current treatments are not successful in eliminating all of the ovarian cancer cells. This may be a result of a protective mechanism of the ovarian cancer cells. Spheroids are large collections of cancer cells that adhere tightly to each other rather than spreading out across an underlying basement membrane. They have been observed in ovarian cancer both in vivo and in ovarian cancer cell lines. Spheroids have been shown to play a role in the metastasis of ovarian cancer cells and have also been shown to be drug resistant. It is our hypothesis that spheroid formation results in epigenetic changes promoting the survival of ovarian cancer cells. Our data suggests that spheroid growth causes the growth rate of ovarian cancer cells to decrease and remain decreased following the transition from spheroids back to monolayer culture. Furthermore, serial passaging of ovarian cancer spheroids results in an increase in resistance to cytotoxic drugs. Since the decrease in growth rate appears to be persistent after spheroid culture it is likely the result of epigenetic modifications that occur during spheroid formation. These epigenetic changes, currently being studied, could become potential drug targets, as new treatments are developed to not only kill ovarian cancer cells but also prevent spheroid formation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1522. doi:10.1158/1538-7445.AM2011-1522
Abstract 1527: Hepatocyte growth factor modulates adhesion of ovarian cancer cells to the peritoneum mesothelium through p70 S6 kinase
Authors: Ip Carman K.M , Yung Susan , Chan Tak-Mao , Wong Alice S.T.
Published: June 2012

Ovarian cancer has the highest mortality of all gynecological cancers. These deaths are mainly due to the highly metastatic characteristic with already widespread peritoneal dissemination and malignant ascites at the time of diagnosis. However the molecular mechanisms that regulate ovarian cancer progression are poorly understood. A key step in this process is the adhesion of ovarian cancer cells to the peritoneal mesothelium. Hepatocyte growth factor (HGF) is present at high concentrations in ovarian cancer ascitic fluid. We show here a role for HGF signaling in the regulation of the peritoneal adhesion of ovarian cancer cells. HGF increased binding of ovarian cancer cells to peritoneal extracellular matrix (ECM) proteins fibronectin and laminin and their receptors α5, α6, and β1 integrin expression. Blockade of HGF resulted in reduced attachment of ovarian cancer cells to these ECM proteins, with a concomitant reduction of α5, α6, and β1 integrin expression, confirming that the effect was HGF specific. Similarly, the peritoneal ECM adhesion could be reversed by neutralizing antibodies to α5, α6, and β1 integrin. The adhesive phenotype and integrin expression activated by HGF could be blocked by specific inhibition of p70 S6 kinase (p70S6K) using short hairpin RNA or small molecule inhibitor, suggesting that HGF transmits the signal through p70S6K. Importantly, it also inhibited adhesion to primary human peritoneal mesothelial cells. These results identify a potential novel mechanism for HGF-induced peritoneal adhesion, which is mediated through p70S6K through regulation of α5, α6, and β1 integrin (This work is supported by the Hong Kong Research Grants Council Grant HKU7599/05M to A.S.T.W.).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1527. doi:10.1158/1538-7445.AM2011-1527
Abstract 1529: Knockdown of cathepsin B and uPAR inhibits CD151 and α3β1 integrin-mediated cell adhesion in glioma
Authors: Malla Rama Rao , Gopinath Sreelatha , Alapati Kiranmai , Gondi Christopher S. , Rao Jasti S.
Published: June 2012

Glioma is a highly complex brain tumor characterized by deregulation of proteins and genes that play important roles in tumor metastasis. CD151, a member of the tetraspanin family of proteins, tightly associates with integrins and accelerates cell adhesion as a modulator of actin cytoskeletal reorganization. Cathepsin B and uPAR are both overexpressed in gliomas and postulated to play central roles in the mediation of glioma metastasis. In the present study, cathepsin B and uPAR were downregulated in U251 and 4910 glioma cells using siRNA plasmid constructs. We evaluated the involvement of CD151 and its associated signaling molecules in adhesion potential. Immunoblot analysis revealed efficient downregulation of cathepsin B and uPAR after siRNA treatments. In addition, siRNA treatments significantly inhibited glioma cell adhesion to laminin as compared to other extracellular matrix proteins. Brain glioma tissue array analysis revealed the expression of CD151 in several tumor samples when compared with normal brain tissue. Further, treatment of glioma cells with cathepsin B and uPAR siRNA led to the downregulation of CD151 and laminin-binding integrins, α3 and β1. Co-localization and immunoprecipitation experiments revealed that downregulation of cathepsin B and uPAR decreased the interaction of CD151 with uPAR/cathepsin B. Transcriptional suppression of CD151 and α3 and β1 integrins using siRNA significantly decreased glioma cell adhesion to laminin. In addition, downregulation of cathepsin B, uPAR, CD151, α3 and β1 significantly decreased the expression of N-cadherin and β-catenin. Studies on the downstream signaling cascade of uPAR/CD151 have shown that phosphorylation of SRC, FAK and Paxillin were reduced with knockdown of cathepsin B, uPAR and CD151. Treatment with the bicistronic construct reduced CD151, α3β1 integrin, N-cadherin and β-catenin expression levels in pre-established glioma tumors in nude mice as well as in tissue lysates. In conclusion, our results show that downregulation of cathespin B and uPAR alone and in combination inhibit glioma cell adhesion by downregulating CD151 and its associated signaling molecules in vitro and in vivo. This study may be useful in dissecting the pathways involved in tumor cell adhesion and for the identification of novel target proteins for therapeutic intervention of gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1529. doi:10.1158/1538-7445.AM2011-1529
Abstract 1532: Small-molecule Stat3 inhibitor sensitizes Cisplatin-resistant ovarian cancer cells to Cisplatin-induced apoptosis
Authors: Sengupta Bhaswati , Startzman A , Turkson J
Published: June 2012

Ovarian Cancer causes the highest number of gynecologic neoplasm related deaths in the United States. The disease and its progression to drug resistance remain poorly understood. While the aberrant activation of the Signal Transducer and Activator of Transcription (Stat) 3 protein is prevalent in ovarian cancer and is detected in chemotherapy resistance, its exact role in the development and maintenance of the drug resistance phenotype remains poorly understood. We sought to investigate the molecular mechanisms by which constitutive Stat3 activity may support Cisplatin resistance in ovarian cancer. Compared to the sensitive A2780S ovarian cancer line, which does not harbor aberrant Stat3 activity, the SKOV3 line harboring constitutively-active Stat3 was partially sensitive to Cisplatin and showed evidence of moderate apoptosis in response to Cisplatin treatment. Compared to the sensitive A2780S cells, the partly-resistant, SKOV3 cells over-express c-Myc, Survivin, and Bcl-xL, and the XIAP anti-apoptotic factor, as well as VEGF pro-angiogenic factor and the matrix metalloproteinases, MMP-2 and MMP-9. The inhibition of constitutively-active Stat3 in SKOV3 cells by the small-molecule inhibitor, S3I-201 suppressed the expression of the anti-apoptotic and the pro-angiogenic factors, and induced moderate degree of apoptosis. Significantly, treatment with S3I-201 enhanced the sensitivity of SKOV3 cells to Cisplatin. Furthermore, the combined treatment of SKOV3 cells with Cisplatin and S3I-201 strongly inhibited colony formation and cell migration in vitro. Our study together identifies potential mechanisms by which aberrant Stat3 activity might support Cisplatin resistance in ovarian cancer, raising the potential that the combination therapy with Stat3 inhibitors would be effective therapeutic and chemo-sensitizing modalities for ovarian cancer and the recurrent disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1532. doi:10.1158/1538-7445.AM2011-1532
Abstract 1538: Inhibition of adenosine-mediated stabilization of intercellular adhesions promotes hypoxia-stressed endometrial carcinoma cell metastasis
Authors: Bowser Jessica L. , Blackburn Michael R. , Shipley Gregory L. , Xie Susu , Broaddus Russell R.
Published: June 2012

Hypoxic microenvironments are commonly encountered in multiple epithelial malignancies. Epithelial cells classically up-regulate cellular adhesive mechanisms to counter the noxious effects of hypoxia. Inhibiting these tissue protective mechanisms is necessary for carcinoma cell invasion and metastasis. Our previous studies utilizing a transgenic model deficient for 5’nucleotidase (CD73), an enzyme responsible for adenosine biosynthesis, demonstrated that adenosine is required to circumvent hypoxia-mediated disruption of cellular adhesiveness in the normal endometrium. In endometrial carcinoma (EC), CD73 is down-regulated in poorly-differentiated, invasive, and metastatic ECs, but is expressed and functional in well-differentiated and non-metastatic disease. Based on these findings, we hypothesized that the malignant progression of EC is dependant on the loss of adenosine-mediated regulation of intercellular adhesions. Modified and unmodified Boyden chambers were used to assess migration and invasion of endometrial adenocarcinoma cells HEC-1A and HEC-1B. Hypoxia (5% or 1.5% O2) significantly reduced HEC-1A and HEC-1B migration and invasion in vitro; this effect was abrogated by CD73 RNA interference and the CD73 enzymatic inhibitor, α, β- methylenediphosphate (AoPCP). The inhibitory effect of CD73 on EC cell migration and invasion was restored by the stable, receptor-targeting adenosine analog, 5’-N-ethylcarboxamido adenosine (NECA). Quantitative RT-PCR identified two adenosine receptors, A1R and A2BR, to be expressed in HEC-1A and HEC-1B. Inhibition of the A2BR by MRS1754 significantly increased HEC-1B migration and invasion, while A1R antagonism (DPCPX) had no effect. A2BR inhibition in HEC-1A monolayers also resulted in increased paracellular permeability to FITC-labeled dextrans. Moreover, A2BR antagonism lowered cellular levels of junctional adhesion proteins claudin-1 and claudin-2, and resulted in increased accumulation of β-catenin within the cytoplasm. Intracellular pools of β-catenin re-organized to the cell membrane with NECA application. In summary, we have shown that CD73-generated adenosine is necessary to maintain normal protective epithelial barrier function in the uterus. Our results demonstrate for the first time that inhibition of adenosine-mediated stabilization of intercellular adhesions promotes cancer cell invasion and metastasis in a hypoxic microenvironment. (NIH TL1RR024147 and NIH 1P50CA098258-01)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1538. doi:10.1158/1538-7445.AM2011-1538
Abstract 1539: Impact of glioma cell gene expression due to semaphorin expression
Authors: Bode Julia , Kietz Silke , Lakomek Max
Published: June 2012

Malignant brain tumours, like the Glioblastoma multiforme, belong to the most frequent tumours encountered in childhood. Unfortunately, even with optimal therapy, consisting of surgical resection and radiotherapy, a glioblastoma multiforme is essentially incurable.

In contrast to the excellent results of well established treatment studies in pediatric patients suffering from leukaemia the cure rates of children with malignant brain tumors remained low. That is why the development of new antitumor strategies has become a major issue in experimental neurology.

Class-3 semaphorins were first identified as glycoproteins that negatively mediate neuronal guidance by binding to neuropilin. The discovery of more widespread expression of class-3 semaphorins in different tumour cell types implicates a role in tumour biology.

The overexpression of class-3 semaphorins in breast cancer cells leads to inhibition of tumour growth. We overexpressed class-3 semaphorins in the human glioma cell line U87MG and monitored a reduction of tumour growth after intracerebral implantation into mice, too. We were also able to detect changes in adhesion and migration. To determine the variation of tumour cell behaviour we analysed the gene expression profile of class-3 overexpressing cells via microarray analysis. We validated changes in transcription levels with the help of realtime-PCR. Changes in the gene expression levels of cell adhesion molecule 2 and Integrin alpha 7 confirmed the changes in adhesion and migration of class-3 semaphorin expressing glioma cells. Further investigations of changes in gene expression levels of different genes are needed to determine the effect of semaphorins on adhesion and migration and the determine the use of class-3 semaphorins as anti-tumour agent.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1539. doi:10.1158/1538-7445.AM2011-1539
Abstract 1544: Identification of microRNA which regulates peritoneal dissemination of ovarian cancer
Authors: Ohyagi Chifumi , Sawada Kenjiro , Morishige Ken-ichiro , Lengyel Ernst , Kimura Tadashi
Published: June 2012

Ovarian cancer is highly metastatic and characterized by widespred peritoneal dissemination and massive ascites, and is the leading cause of death from gynecological malignancies. We have previously identified integrin α5 as a promising target for ovarian cancer treatment by inhibiting peritoneal dissemination. Our aim is to newly identify microRNAs which regulate integrin α5 expression and analyze the therapeutic potential of targeting those microRNAs.

Based on the microRNA algorism search, we identified hsa-mira-92a as one of the candidates. Hsa-mir-92a is little expressed in ovarian cancer cells which express high level of integrin α5, whereas hsa-mir-92a is highly expressed in RMUG-S cells which hardly express integrin α5. Forced expression of mir-92a in ovarian cancer cell lines reduced integrin α5 expression in mRNA and protein level, accompanied by the inhibition of cell adhesion and invasion. On the contrary, the antagonist of mir-92a enhanced integrin α5 expression in RMUG-S cells. Luciferase assay revealed that mir-92a reduces the transcriptional activity of integrin α5 by directly binding ITGA5 3′-UTR. In vivo ovarian cancer xenografts, transducing hsa-mir-92a expressing plasmid into ovarian cancer cell lines significantly reduced peritoneal dissemination and ascites formation.

Those results suggested targeting hsa-mir-92a could be a promising therapy for a subset of ovarian cancer patients by inhibiting integirn α5 expression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1544. doi:10.1158/1538-7445.AM2011-1544
Abstract 1547: Characterization of receptor-ligand interactions between head and neck circulating tumor cells and E-selectin
Authors: Marshall Jocelyn R. , Reynolds Nathan M. , Abram Courtney E. , Henson Karissa A. , Wood S. Matthew , Nimrichter Leonardo , Burdick Monica M.
Published: June 2012

Though the exact mechanisms of cancer metastasis are not known, it is widely believed that circulating tumor cells (CTCs) that have broken off from the primary tumor flow through the blood stream until ligands on their surface recognize receptors present on the vascular endothelium. From there, CTCs loosely attach to the endothelium surface and roll until they become firmly adherent. Cells then migrate through the blood vessel walls to establish metastatic colonies in a distant organ. Therefore, identifying the molecular structures available for adhesion on the surface of the CTCs is compulsory to fully understanding the ubiquitous and unique natures of cancer metastasis. We have previously shown that the adhesion of head and neck squamous cell carcinomas (HNSCCs) to human umbilical vascular endothelium cells (HUVECs) under flow is mediated by E-selectin present on the endothelium surface. In this study, we investigated the interaction of E-selectin with ligands present on the HNSCC surface. Protease-treated cells of the JHU-011-SCC, JHU-013-SCC, and FaDu cell lines perfused through the parallel-plate flow chamber tethered to and rolled on the HUVECs at a greater incidence rate compared to untreated control cells. These binding interactions were sialidase and fucosidase sensitive. Altogether, these results indicate that sialofucosylated glycolipids on HNSCC cells predominantly contribute to the adhesion of these cells to endothelium. This observation was further confirmed by probing western blots of SDS-PAGE resolved HNSCC lysates with HECA-452 mAb and E-selectin chimera. Neither probe was reactive with protein species. We also tested if a shielding effect of hyaluronic acid (HA) prevented glycosphingolipid engagement to E-selectin. When HNSCC cells were treated with HAse and when hyaluronic acid synthase-3 was silenced by shRNA, there was an increase in cell adhesion events. These results clearly demonstrate that proteins and HA exert shielding effects on the E-selectin glycolipid ligands, thus affecting the overall metastatic ability of the HNSCCs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1547. doi:10.1158/1538-7445.AM2011-1547
Abstract 1548: The effects of synthetic triterpenoids on GSK3β and focal adhesions
Authors: To Ciric , Di Guglielmo John M.
Published: June 2012

Synthetic triterpenoids are a class of anti-cancer compounds that are efficacious in targeting multiple cellular functions including apoptosis, growth inhibition, anti-inflammation and cytoprotection in both cell culture and animal tumor models. However, its effects on cell migration, a precursor event to cancer metastasis, remain poorly understood. Previously, we have shown that the methyl ester derivative of 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO-Me) inhibits cell migration. The objective of our current study is to examine in further detail the mechanism by which CDDO-Me blocks cell migration. Using various proteomic approaches as well as affinity pull down assays, Glycogen Synthase Kinase 3 Beta (GSK3β) was identified and confirmed as a triterpenoid-binding protein. Since GSK3β has been shown to be an important regulator of cell adhesion, a process that is intricately connected to cell migration, we hypothesize that CDDO-Me may target GSK3β and affect cell adhesion dynamics to inhibit cell migration. Indeed, we have observed that triterpenoid treated cells have decreased GSK3β activity. We also observed that the size and shape of focal adhesions were altered in the presence of synthetic triterpenoids. The molecular link between synthetic triterpenoids, GSK3β and focal adhesions are currently being investigated to better describe how this promising anti-cancer compound may act via GSK3β to affect focal adhesions and cell migration.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1548. doi:10.1158/1538-7445.AM2011-1548
Abstract 1549: Regulation of inflammation and tumor growth by the novel XL313 cryptic ECM element
Authors: Ames Jacquelyn J. , Nugent Desiree P. , Contois Liangru , Caron Jennifer M. , Brooks Peter C.
Published: June 2012

Experimental evidence suggests that multiple cell types within the tumor microenvironment such as fibroblast, endothelial cells and inflammatory cells contribute to tumor growth. Recent evidence indicates that biomechanical alterations in the structural integrity of ECM proteins may result in exposure of cryptic regulatory elements that play roles in differentially controlling cell signaling pathways. Therefore, exposure of cryptic regulatory sites may function as biomechanical switches, allowing various cell populations to respond in distinct ways to localized structural alterations within the tumor microenvironment. Thus, identifying and characterizing biomechanical ECM switches may provide selective new therapeutic targets to regulate tumor progression. In this regard, triple helical collagen has numerous RGD containing cryptic integrin binding sites. However, it is not known whether all these cryptic RGD elements are functionally redundant or whether distinct flanking sequences may convey differential control of cellular behavior.

To examine potential functional differences between distinct RGD containing cryptic sites, we generated a series of Mabs directed to synthetic RGD containing peptides found within type-I collagen. One Mab termed XL313 selectively recognized a cryptic element containing the amino acid sequence RGDKGE, but failed to bind other RGD containing sites. Mab XL313 failed to recognize intact triple helical collagen, but did bind proteolytically cleaved collagen. Importantly, Mab XL313 significantly (P<0.05) inhibited B16F10 melanoma tumor growth by greater than 50%. Given the possibility that inflammation may contribute to tumor development, we examined the impact of the XL313 epitope on growth factor induced inflammation. Our studies suggest that the XL313 cryptic site may regulate inflammation as Mab XL313 significantly (P<0.05) inhibited inflammation / CAM thickening in the chick embryo model. Collectively, our studies suggest that the XL313 biomechanical ECM switch may represent an important new therapeutic target for the control of tumor growth and inflammation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1549. doi:10.1158/1538-7445.AM2011-1549
Abstract 1555: Immune suppression of regulatory T cells and M2 macrophage in breast cancer patients
Authors: Imoto Shigeru , Sakemura Noriko , Ito Hiroki , Imi Kentaro , Isaka Hirotsugu , Tazaki Eri , Kanma Hiroshi , Wada Noriaki , Nakatsura Tetsuya
Published: June 2012

Background: Recent multi-disciplinary treatments of breast cancer (BC) could decrease the mortality rate, but successful immune therapy remains uncertain. To explore new strategy of immune therapy of BC, we are investigating about host-tumor immune response in BC patients. Materials and methods: Tumor tissue specimens and peripheral blood mononuclear cells (PBMC) were obtained from early and advanced BC patients. PBMC were also collected from healthy volunteers. Regulatory T (Treg) cells were examined by counting CD4+CD25highCD127low/-cells in PBMC with flow cytometry analysis. Immunohistochemical evaluation of tumor specimens was performed with monoclonal antibodies of HLA-ABC and DR, CD56, CD68, CD83, and CD163. The number of stained cells was analyzed using a semiquantitative ordinal scale ranging from 0 to 3 (0, +/-, ++, +++). Results: HLA type I and type II were stained negative in 44% and 82% of 50 BC cases. When host-tumor immune response were compared between 38 early BC cases and 12 advanced BC cases, numbers of CD68-positive cells significantly increased in peripheral tumor tissues of advanced BC cases than in those of early BC cases (92% versus 53%). CD163-positive tumor cells were also detected more frequently in advanced BC cases than in early BC cases (75% versus 16%). There were no significant differences of distribution of CD4, CD8, CD56, and CD83-positive cells in early and advanced BC cases. Treg cells in PBMC significantly increased in percentage of the population in 46 BC patients than in 16 healthy volunteers (3.8% versus 2.1% of CD4-positive cells at mean value). Interestingly, Treg cells decreased in percentage of the population in postoperative BC patients. Conclusions: Our results suggest that Treg cells render BC patients under immune suppression and M2 macrophage plays an important role of tumor progression. Targeted therapy against M2 macrophage traits may be a promising strategy of breast cancer. Host-tumor immune response will be reported for BC patients treated with primary chemotherapy or radiofrequency ablation therapy in comparison with those who underwent breast surgery first.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1555. doi:10.1158/1538-7445.AM2011-1555
Abstract 156: MicroRNA expression profiling of human esophageal metaplastic changes
Authors: Fassan Matteo , Volinia Stefano , Palatini Jeff , Pizzi Marco , Fernandez-Cymering Cecilia , Battaglia Giorgio , Balistreri Mariangela , Croce Carlo M. , Zaninotto Giovanni , Ancona Ermanno , Rugge Massimo
Published: June 2012

For nearly the last two decades, the diagnosis of Barrett's mucosa (BM) strictly required histologically documented intestinal metaplasia (IM). More recently, a precancerous potential has been also associated to non-goblet columnar metaplasia (i.e. gastric metaplasia, [GM]). Molecular profiling can be applied to characterize the precancerous meaning of non-intestinal versus intestinal metaplastic transformation of the native squamous esophageal mucosa. MicroRNAs (miRNAs) are a class of small non-coding RNAs controlling gene expression and they are reportedly dysregulated in Barrett's carcinogenesis.

To investigate the role of miRNAs in the natural history of esophageal metaplasia, miRNA microarray analysis (OSU-CCC version 4.0; Ohio State University) was performed in a series of 30 matched biopsy samples (10 native squamous epithelium [N], 10 GM and 10 IM) from 10 long-segment BE-patients. Microarray findings were validated by qRT-PCR using another series of 20 consecutive BE-patients.

miRNA profiling consistently disclosed different metaplasia-specific miRNA signatures for N versus GM, and N versus IM. Comparing the two signatures disclosed that they had 11 significantly dysregulated miRNAs in common, 8 of which were proportionally dysregulated in the sequence N-GM-IM (2 overexpressed [hsa-miR-192 and hsa-miR-215] and 6 underexpressed [hsa-miR-18a*; hsa-miR-20a, hsa-miR-106a, hsa-miR-203, hsa-miR-205 and hsa-miR-769-5p]).

Specific miRNAs are involved in the natural history of BE. Molecular profiling of phenotypical lesions included in Barrett's carcinogenesis supports the hypothesis that squamous epithelium is converted initially into non-goblet columnar epithelium, before goblet cell metaplasia sets in.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 156. doi:10.1158/1538-7445.AM2011-156
Abstract 1562: The G protein-coupled receptor mFPR2 promotes antitumor host defense
Authors: Liu Ying , Chen Keqiang , Yao Xiaohong , Gong Wanghua , Yoshimura Teizo , Ford Jill , McVicar Daniel , Wang Ji Ming
Published: June 2012

mFPR2 is a G protein-coupled receptor interacting with chemotactic agonists produced by bacteria and the host. In vivo, mFPR2 plays a major role in innate and adaptive immune responses as shown by reduced severity of allergic airway inflammation and diminished production of Th1 and Th2 cytokines in mFPR2-deficient mice. These defects were associated with impaired trafficking of antigen presenting dendritic cells into the inflammatory sites and the draining lymph nodes. The aim of this study is to examine the role of mFPR2 in tumor progression. We found that mFPR2-deficient mice bearing subcutaneously implanted Lewis Lung Cancer cells (LLC) exhibited significantly shortened survival rate due to more rapidly growing tumors. In contrast, in mFPR2-transgenic (Tg) mice, subcutaneously implanted LLC tumors grew more slowly than those in wild type (WT) littermates. We further observed a decrease in the proportion of CD11b+ and Gr1+ myeloid cells accompanied by reduced infiltration of CD11b+Gr1low myeloid-derived suppressor cells and CD4+Foxp3+ regulatory T cells in LLC tumors in mFPR2-Tg mice. These results suggest that mFPR2 may play an important role in anti-tumor host defense by down-regulating immune suppressive cell infiltration in tumor tissues.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1562. doi:10.1158/1538-7445.AM2011-1562
Abstract 1567: Pancreatic tumor cell secreted Cyr61/CCN1 enhances abnormal angiogenesis: A key event for metastasis
Authors: Mehta Smita , Majumder Monami , Haque Inamul , Banerjee Snigdha
Published: June 2012

CYR61/CCN1, a member of the cysteine rich 61/connective tissue growth factor/nephroblastoma over expressed (CYR61/CTFG/NOV) family of growth factors, is a pro-angiogenic factor that mediates diverse roles in development, cell proliferation, and tumorigenesis. In our previous studies, we have demonstrated that Cyr61 is highly expressed in aggressive pancreatic cancer cell lines. The objective of this study is to evaluate whether tumor cell-secreted Cyr61 is able to enhance the interactions of endothelial cells and vascular smooth muscle cells for remodeling of vasculature. To do so, we determined the impact of conditioned media (CM) of different pancreatic cancer cell lines and their side populations on in vitro and in vivo angiogenesis. The in vitro angiogenesis assay demonstrates that the CM of Cyr61 positive pancreatic cancer cells drastically altered the angiogenesis. CM induces deformed capillary-like structures as compared to regular media and this effect is cell type specific. Moreover, CM also drastically altered the attachment of vascular smooth muscle cells to the capillaries. These in vitro studies were further confirmed by in vivo gel foam assay in mice. The abnormal effects of CM can be overruled by nullifying the Cyr61 expression through Cyr61-specific shRNA or neutralizing Cyr61 antibody. Collectively, these studies suggest that Cyr61 could be a target molecule to normalize the angiogenesis to prevent metastatic growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1567. doi:10.1158/1538-7445.AM2011-1567
Abstract 1576: Prostaglandin I2 signaling regulates micro-metastasis in lung cancer
Authors: Minami Yoshinori , Okumura Shunsuke , Sasaki Takaaki , Kawabe Junichi , Endo Satoshi , Satoh Kazuhiro , Kitada Masahiro , Fujita Yuka , Hasebe Naoyuki , Ohsaki Yoshinobu
Published: June 2012

Background: Bone marrow-derived cells (BMDCs) play an important role in the initiation of neo-angiogenesis, and contribute to the diseases including ischemic diseases, retinopathy, and cancer. We have reported that BMDCs express prostaglandin I2 (PGI2) specific receptor, IP and the PGI2-IP signaling system maintains the function of BMDCs. Additionally, we have reported that the PGI2-IP signaling in BMDCs play an important role for tumor growth, by using mouse bone marrow transplanted models and tumor xenograft mouse models.

Objectives: The purpose of this study was to test the hypothesis that PGI2-IP signaling system regulates micro-metastasis in lung cancer.

Methods: In vitro study, 5-bromo-2’-deoxy-uridine (BrdU) cell proliferation assay and clonogenic cell assay were used to clarify the effect on lung cancer cell (A549, PC9) proliferation and cellular form in the existence of IP receptor agonist or antagonist. Human cytokine antibody array was performed to detect the expression of angiogenic cytokines and growth factors secreted by the treated cells.

In vivo study, we employed both a mouse lung metastasis model and a mouse bone marrow transplantation model. We used GFP-labeled or DsRed-labeled lung cancer cells to distinguish cancer cells from BMDCs. Lung cancer cells were injected into the tail vein of mice (ICR nu/nu), which were transplanted with bone marrow cells from GFP-labeled wild-type or IP knockout mice. Real-time RT-PCR was used to quantify lung metastasis of human cancer cells in nude mice, and the expression levels of housekeeping genes (human and mouse GAPDH) in the lungs were measured. Immunofluorescence assay was performed to evaluate angiogenesis in pulmonary metastatic tumors.

Results: In vitro study, PGI2-IP signaling didn't impair proliferation of lung cancer cells and production of angiogenic cytokines secreted by tumors.

In the mice lung metastasis model, micro-metastasis in lung cancer was regulated by PGI2-IP signaling system. RT-PCR assay showed that PGI2-IP signaling changed the expression levels of human lung cancer cells in mice lungs. In the immunofluorescence assay, PGI2-IP signaling influenced the expression of pericyte markers around the tumors.

Conclusions: The present study demonstrated that PGI2-IP signaling system regulates micro-metastasis in lung cancer. These results suggest that PGI2-IP system may become a novel therapeutic target in lung metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1576. doi:10.1158/1538-7445.AM2011-1576
Abstract 1577: The Impact of M1/M2 Tumor Associated Macrophage (TAM) Polarization on Cancer Progression in NSCLC
Authors: Hsiao Yi-Jing , Yu Sung-Liang , Chen Huei-Wen , Chen Jeremy J.W. , Yuan Ang , Yang Pan-Chyr
Published: June 2012

Tumor microenvironment has important influences on tumor progression. One of the effectors is tumor-associated macrophages (TAMs) that are characterized as M1/M2 by their different immune responses, induced by Th1 and Th2.

In our study, we polarized THP-1 into classically (M1) and alternatively (M2) activated macrophages and evaluated some different markers to distinguish M1 and M2 subtype (M2a, M2c). M1 macrophage expressed high IL-1, IL-6, IL-12, IL23, TNF-α, and a surface maker CCR7; M2a and M2c macrophages expressed high IL-10 and a surface maker CD206, but expressed CD23 and CD163, respectively. Microarray analysis was used to evaluate the alteration of gene expression of lung cancer cell line (A549) after coculture with different subtype TAMs (M0, M1, M2a and M2c). Bioinformatics analysis indicated that TAMs mainly alter cell cycle, coagulation and cell adhesion pathways in tumor cells.

The M0, M2a and M2c promoted the invasion ability of A549 after coculture for 2days. Otherwise, M1 condition medium reduced cell viability and induced apoptosis of A549 but not H1299, the p53 null cell line. Moreover, the M2 macrophage condition medium but not M1 increased the drug resistance ability of A549 cells after long-term coculture. More importantly, in vivo tumorigenesis assay indicated that M2 can promote tumor growth and angiogenesis. Different subtypes of macrophages can modify behaviors of cancer cells and may contribute to tumor progression. Further studies in the detailed mechanisms involved in the interaction of TAMs and cancer cells will unveil the role of macrophages in tumor progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1577. doi:10.1158/1538-7445.AM2011-1577
Abstract 1582: Identification of miRNAs that regulate breast cancer stem cells and spontaneous metastases in orthotopic mouse models
Authors: Liu Huiping , Bockhorn Jessica , Dalton Rachel , Olopade Olufunmilayo F. , Clarke Michael F. , Greene Geoffrey L.
Published: June 2012

To examine the role of microRNAs (miRNAs) in breast cancer progression, we profiled miRNA expression in both clinical breast tumors and human-in-mouse breast tumor models, where breast cancer stem cells (BCSCs) contribute to spontaneous metastasis. CD44+ cells from both primary tumors and lung metastases were highly enriched for tumor initiating cells. Based on the miRNA profile analyses, we identified a limited number of miRNAs that are differentially expressed in metastatic triple-negative breast tumors and that regulate BCSCs and tumor invasion in vitro. To facilitate miRNA functional studies in vivo, we also developed tumor imaging approaches by transducing BCSCs with optical reporter fusion genes (Luc2-eGFP or -tdTomato), which enabled both bioluminescence imaging (BLI) and FACS-based analysis and sorting. With non-invasive BLI approaches, as few as 10 cells of stably labeled BCSCs can be tracked in vivo. When optical reporters are expressed along with miRNA precursors or inhibitors, the effects of introduced miRNA candidates can be evaluated by selective imaging of the transduced tumor cells, thereby eliminating the noise of un-transduced cells. Using this model system and imaging technology, we have screened and identified miRNAs that regulate BCSCs (e.g. miR-200 family) and metastatic CSCs (MCSCs). Our metastatic cancer models, combined with non-invasive imaging techniques, comprise an integrated approach that could be applied to dissect the molecular mechanisms underlying the dissemination of MCSCs, to explore therapeutic strategies targeting metastasis in general, or to evaluate individual patient tumor cells and predict response to therapy.

Supported in part by the NIH T90, DOD BC087695 and Chicago Fellows Program at The University of Chicago (H.L.), University of Chicago Cancer Research Center Pilot Research Fund, UCMC/Northshore Collaborative Research Award and Ludwig Fund (G.L.G and H.L). NIH R01 and Breast Cancer Research Foundation (M.F.C. and H.L.).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1582. doi:10.1158/1538-7445.AM2011-1582
Abstract 1594: Significant in vivo activity of ENMD-2076, a novel inhibitor of Aurora receptor tyrosine kinases, alone and in combination with lenalidomide against plasma cell myeloma
Authors: Sandusky George E. , Rodgers Matt , Zhang ShuHong , Sinn Anthony , Pollok Karen , Farag Sherif
Published: June 2012

Aurora kinases, particularly aurora A and aurora B, play a role in normal cell proliferation. Since an unusually high expression of Aurora kinases has been reported in tumor cells, they have become the target of a variety of novel anticancer drugs. One such drug, ENMD-2076, was studied in a translational research project involving a xenograft mouse model and a phase I clinical trial. The preclinical xenograft model evaluated multiple myeloma tumors in NOD/SCID mice treated with ENMD-2076, lenalidomide, and combinations of both drugs, and compared to an untreated vehicle control group. Immunohistochemistry (IHC) was used to predict the effectiveness of treatment for multiple myeloma based on caspase-3, Ki67, phospho-histone H3 (p-H3), and CD34 biomarker expression. IHC slides were quantified using both a traditional pathology hand count and an image analysis program (Image J). The Image J data closely mirrored the hand count for all four biomarkers. At the highest combined dosage, hand count data showed CD34 levels down 74.2% from the vehicle control. Ki67 and p-H3expression were down slightly, and caspase-3 was increased two fold in the combined drug treatment group. Comparing the two drugs, it was evident that the ENMD-2076 aurora kinase inhibitor had a stronger anti-angiogenic effect, while the Lenalidomide had a stronger apoptotic effect. Although the human clinical trial is still in its early stages, initial trends are consistent with results seen in this mouse model.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1594. doi:10.1158/1538-7445.AM2011-1594
Abstract 1602: Mucinous adenocarcinoma developed from human fallopian tube epithelial cells through a combination of defined genetic modifications and tumor microenvironment
Authors: Mercado-Uribe Imelda , Shan Weiwei , Liu Jinsong
Published: June 2012

Epithelial ovarian cancer includes several distinct histologic types including serous, mucinous, endometrioid, and clear cell. The cell origin of these distinct epithelial ovarian cancers has been debated for many years. Recent studies have suggested that some ovarian and pelvic serous carcinomas could originate from the fimbrial end of distal fallopian tube, although direct experimental evidence supporting this hypothesis is still lacking. To test this hypothesis, we immortalized a normal human fallopian tubal epithelial (FTE) cell line by using a retrovirus-mediated infection expressing the early region of SV40 T/t antigen and the human telomerase reverse transcriptase. These immortalized FTEs were then transformed by ectopic expression of oncogenic human HRASV12. Tumorigenicity of the immortalized and/or transformed cells was subsequently tested by anchorage-independence assay and inoculation into nude mice via subcutaneous or intraperitoneal injections. As expected, the HRASV12-transformed FTEs produced tumors through both subcutaneous and intraperitoneal injections, while no tumor growth was observed in immortalized FTEs. To our surprise, however, while histopathologic examination of resulting tumor from subcutaneous injections revealed a largely undifferentiated carcinoma with focal mucin production, tumors from intraperitoneal injections showed mucinous adenocarcinoma with rich mucin production from an undifferentiated area. The mucinous differentiation of tumors was most prominent near the peritoneal fat and other peritoneal organs including the pancreas, spleen, and intestine. Immunohistochemical staining of tumor cells showed positive staining for the epithelial cell markers cytokeratin AE1/AE3 and CA125. Our study demonstrates that FTEs can generate mucinous adenocarcinoma through genetic modifications and that such mucinous differentiation is dependent on the peritoneal microenvironment. Thus, the experiment described here provided the first experimental evidence that fimbrial epithelial cells of the fallopian tube could be a potential source of ovarian mucinous adenocarcinoma, a distinct histologic entity of epithelial ovarian cancers whose origin is largely unknown.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1602. doi:10.1158/1538-7445.AM2011-1602