bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5463: Serum aminotransferases and alkaline phosphatases as biomarkers of hepatotoxicity in sprague-dawley rats exposed to silver nanoparticles
Authors: Patlolla Anita K. , Tchounwou Paul B.
Published: August 2012

Nanoparticles are small scale substances (<100 nm) used in biomedical applications, electronics, and energy production. Increased exposure to nanoparticles being produced in large scale industry facilities elicits concerns for the toxicity of certain classes of nanoparticles. In particular, the oral toxicity of silver nanoparticles is of concern to ensure public and consumer health. Due to the intensive commercial application of silver nanoparticles (Ag-NPs), health risk assessment of this nanoparticle is of great importance. The previous in vitro studies demonstrated that Ag-NPs caused toxicity in various cell-lines. However, toxicity of Ag NPs in vivo is largely lacking. This study evaluated the effect of Ag-NPs on the activities of specific liver enzymes such as aminotransferases (GOT/GPT), and alkaline phosphatases (ALP) which may be useful as biomarkers of hepatotoxicity. Four groups of five male rats each weighing approximately 80 + 2 g were orally administered once a day for five days with doses of 5, 25, 50 and 100 mg/kg BW of silver nanoparticles. A control group was also made of 5 rats. At the end of the experiment, serum samples were collected following standard protocols. The data obtained from the hepatotoxicity study clearly show that highest two dose 50 and 100 mg/kg of silver nanoparticles has statistically significantly increased the activity of serum aminotransferases (GOT and GPT) and alkaline phosphatases (ALP) when compared to control. The results demonstrate that Ag-NPs have the potential to induce hepatotoxicity in Sprague-Dawley rats. Our result does not imply that silver nanoparticles should be banned from use but more in vivo studies with histopathological characterization should be designed to confirm the results of this study.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5463. doi:1538-7445.AM2012-5463
Abstract 555: A role for XMRV in prostate tumor promotion in vivo
Authors: Comeaux Brennen P. , Zolochevska Olga , Figueiredo Marxa
Published: August 2012

Xenotropic murine leukemia virus-related virus (XMRV) is a retrovirusclosely related to murine leukemia viruses, which cause a variety of cancersand immunological and neurological diseases in mice. Whether XMRV plays a rolein cancer and other diseases remains controversial; some studies show apossible association, but there is currently no evidence that the virus is acausal agent for disease. Our hypothesis was that the 3′ long terminal repeat(LTR) from the XMRV genome is responsible for its oncogenic activity inprostate cells. We chose three regions from the XMRV 3′ LTR and tested thepotential of these regions in transactivating a cellular gene promoter whosehomolog is transactivated by MMLV in mouse cells. We examined thetransactivation potential in pre-cancerous and a panel of cancerous prostateepithelial cells. Our results showed that for certain 3′LTR regions inducenearly full transactivating activity, while others might comprise inhibitoryregions. Therefore, to better understand the mechanism by which the XMRV 3′ LTRregions might be inducing or inhibiting transactivation, we tested theirability to transactivate cellular genes related to transformation. We used asreporters plasmids containing binding sites for cellular transcription factors.The results showed several similarities in transactivation, suggesting commonactivation pathways for 3′LTR transactivating activity. Finally, we tested ifXMRV infection could enhance oncogenicity potential of prostate precancerousand cancerous cell lines. We examined this in vitro by assessing the rate of colonyformation in soft agar, and in vivo, using xenograft assays. The resultssuggested that cell lines infected with XMRV had enhanced in vitro colonyformation, as well as enhanced in vivo growth, however, addition of anothertumorigenic genetic alteration was required in conjunction with XMRV-infectionto induce enhanced tumor growth. Our data suggests that the XMRV 3′LTR may havea transactivating activity on cellular genes, and that XMRV stimulatepromotion/progression and not ‘initiation’ of tumorigenesis. These results arenovel since they represent for the first examinations of the role of XMRV intumorigenesis of prostate epithelial cell lines in vitro and in vivo andexaminations of the 3′LTR context of the virus upon transactivation of cellulargenes as a potential mechanism of cellular transformation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 555. doi:1538-7445.AM2012-555
Abstract 5550: Ingenol mebutate induces an early and inflammatory wound healing response in a human in vitro 3-D skin model
Authors: Litman Thomas , Høyer-Hansen Maria H. , Zibert John R.
Published: August 2012

Ingenol mebutate (IM, ingenol-3-angelate, PEP005) is a promising new agent for topical treatment of actinic keratosis (AK), a pre-malignant condition that if left untreated can develop into squamous cell carcinoma. A dual mechanism of action of IM has been suggested; this involves direct lesional cell death and PKC activation dependent, immune modulatory effects. The aim of this study was to investigate the mode of action of IM. We have previously compared IM's penetration through normal skin to a 3-D reconstructed human skin (RHS) model (EpiDermFT™, MatTek). A 10-fold higher permeability was found in RHS, which likely reflects the increased permeability of AK lesions. Therefore, we studied the effects of 0.05% IM gel in this model at different time points (0.5, 6, 24, and 96 h). The response parameters measured included cytotoxicity (MTT assay), cytokine secretion, as well as global transcriptomic profiling (Affymetrix GeneChip® Human Gene 1.0 ST arrays). Multivariate analysis, including principal component analysis, unsupervised hierarchical clustering, and gene ontology enrichment, revealed that the major effects of IM comprised: cell death, inflammation and a wound healing response. This was indicated by differential expression of e.g. CASP14, HAS2, several MMPs, SERPINs, ILs, and CCL20. We also observed regulation of many skin-specific genes, related both to epidermis development, such as keratinocyte differentiation, and to fibroblasts in dermis, reflecting the nature of the RHS. Furthermore, a transcription factor binding site (TFBS) analysis suggested ERK2 and PKC as relevant targets for IM. This is in accord with the drug's transient immune modulatory effect. Indeed, a specific, inflammatory response was observed already after 30 min treatment, and several genes, including MMP10 and IL1R2, displayed a time-dependent increase in expression, reaching a maximum at 24h returning to normal at 96h. A sustained effect was observed for more than 300 genes, most of which were down-regulated following IM treatment. In conclusion, IM displays pleiotropic effects, of which many are regulated on a transcriptional level. Our data suggest transient activation of in particular PKC and ERK2, with a link to cytokine release and wound healing response. The inflammatory reaction may subsequently contribute to the cell death observed after 24h treatment with IM.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5550. doi:1538-7445.AM2012-5550
Abstract 5551: Somatic mutations in KRAS, BRAF and PIK3CA genes in Chilean patients with sporadic colorectal cancer: Correlation with clinical and histological features
Authors: Hurtado Claudia , Zarate Alejandro , Wielandt Ana María , Carrillo Katya , Yamagiwa Ken , Kronberg Udo , Levican Jorge , Contreras Luis , Lopez-Kostner Francisco
Published: August 2012

Background: KRAS, BRAF and PIK3CA proto-oncogenes encode for proteins involved in epidermal growth factor receptor (EGFR) signaling pathways. Activating mutations in these genes may contribute to tumor development and induce resistance to biological therapies in patients with metastatic colorectal cancer (CRC). Furthermore, microsatellite instability (MSI) has been proposed as a prognostic and predictive marker in sporadic CRC. Patients and methods: In tumors of patients undergoing surgery for sporadic CRC at our institution, somatic mutations in KRAS, BRAF and PIK3CA genes were assessed by single-strand conformation polymorphism(SSCP) and direct sequencing. MSI analysis was performed using the 5 markers of the standard NIH panel. Clinical and histopathological data of the patients were obtained from our prospectively maintained database. Results: A total of 58 patients with a nearly equal gender distribution and a mean age of 62 years were included. A total of 31 mutations in 26 patients were identified. The frequency of mutations was 28% for KRAS, 9% for BRAF and 17% for PIK3CA. No difference was found in the mutation frecuency between the different tumor locations (42,1% in right colon vs. 46,2% in left colon/rectum, P= NS) or lymphnode status (37,5% in node positive vs. 50% in node negative tumors, P=0,346). Tumors with deeper invasion showed a higher frecuency of mutations, but this tendency did not reach statistical significance (32% in pT1-2 vs. 51% in pT3-4, P=0,157). In 26% of the patients, MSI-high was observed, more frecuently in tumors of the right colon (57,9% vs 10,3%, P<0,001). Patients with BRAF mutations, a higher proportion of MSI-high was found compared to BRAF wildtype (27% vs. 2,3%, P=0,013). There was no correlation between the lymphnode positivity and MSI status. Conclusion: Our results show that the frequency of mutations in patients with sporadic CRC in Chile is similar to that described in other countries. The presence of MSI-high is correlated with tumors in the right colon and with mutations in the BRAF gene. The tendency of finding more mutations in locally advanced and node-negative tumors needs to be confirmed in future studies with a larger number of patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5551. doi:1538-7445.AM2012-5551
Abstract 5552: Macrophages increase brain metastasis in a chick embryo model
Authors: Menen Rhiana , Kolostova Katarina , Bobek Vladimir , Hassanein Mohamed K. , Bouvet Michael , Hoffman Robert M.
Published: August 2012

Macrophages are known to promote tumor growth by various mechanisms. In the present study, we determined if macrophages could promote metastasis in the chick embryo models. The chorioallentoic membrane (CAM) of six chicken embryos was inoculated with 106 PC-3 human prostate cancer cells stably expressing GFP. Twelve hours later, 106 macrophages isolated from the peritoneal washings of three nude mice were added to the CAMs of three embryos. Embryo brains were removed on day 8 post-inoculation, and processed for cryosectioning. Imaging was performed using the IV-100 scanning laser microscope (Olympus Corp, Tokyo, Japan). Images were processed using Image J software in order to calculate the number, size, and total area of brain metastases in the control and macrophage groups. The macrophage group resulted in an average of 43.3 brain metastases compared to 5.5 in the control group (p=0.000001). There was also a significantly greater total metastatic area in the macrophage group of 504.8 mm2 compared to 100.5 mm2 in the control group (p= 0.000001). However, the control group showed a significantly larger size of individual metastases of 28.9 mm2 compared to 19.3 mm2 in the macrophage group (p= 0.0007). Macrophages, therefore, promote brain metastasis in the chick embryo model of human prostate cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5552. doi:1538-7445.AM2012-5552
Abstract 5559: Detection of IDH1 mutation in the plasma of glioma patients
Authors: Boisselier Blandine , Labussiere Marianne , Rossetto Marta , Larraya Jaime Gallego Perez , Marie Yannick , Delattre Jean-Yves , Sanson Marc
Published: August 2012

Background IDH1 mutation is both a diagnostic and a strong prognostic factor in gliomas. In this work, we investigated the feasibility of detecting the IDH1 mutation in plasma of glioma patients. Patients and Methods The study was conducted in 31 controls and 80 glioma patients with known IDH1 status, and confronted with MRI data. Small Size DNA (ssDNA) was extracted from 4mL plasma and the IDH1R132H mutation was detected by a combination of COLD (co-amplification at lower denaturation temperature) and digital PCR. Results ssDNA concentration (150pb to 200pb) ranged from 0.1 to 6.6 ng/mL (median 1.2 ng/mL) in controls and 0.1 to 50.3 ng/mL (median 1.2 ng/mL) in patients. ssDNA concentration did not correlate with tumor volume in low grade glioma. In contrast, ssDNA concentration in high grade gliomas patients was significantly higher than in healthy controls and correlated with the enhancing volume V (V=0: 0.9 ng/ml [0.1-3.5]; V< 10 cm3: 1.6 ng/ml [0.4-24.9]; V>10 cm3: 14.0 ng/ml [0.6-50.3]). We then analyzed 39 patients (25 with R132H IDH1 mutated glioma, 14 with IDH1 wild type glioma) for the detection of IDH1 mutation on the ssDNA extracted from plasma. The mutation was detected in plasma DNA of 15 out of 25 patients (60%) with IDH1 mutated tumor, and in none of the 14 patients with IDH1 wild type tumor. The detection rate of IDH1 mutation in plasma was higher in high grade (12/17 patients (70.6%) and was correlated with the enhancing tumor volume. Interestingly all the mutant tumors with an enhancing tumor volume higher than 3.5 cm3 (9/9) were detected by digital PCR in plasma. Conclusions Our study shows that (i) tumor DNA is released in glioma patients; (ii) ssDNA concentration is correlated with enhancing tumor volume, and (iii) IDH1 mutation is detectable in plasma DNA. This represents the first step for developing a quantitative plasma biomarker that could be used to monitor glioma status.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5559. doi:1538-7445.AM2012-5559
Abstract 5560: PROX1 expression and function in malignant gliomas
Authors: Elsir Tamador , Qu Mingqi , Berntsson Shala , Olofsson Tommie , Ribom Dan , Orrego Abiel , Hedren Anna , deimling Andreas V. , Hartmann Christian , Lindström Mikael , Smits Anja , Nister Monica
Published: August 2012

The PROX1 transcription factor, also known as Prospero in flies, plays a critical role in the development of various organs especially the lymphatic and central nervous systems. Mutations in Prospero induce brain tumor development in flies due to the lack of normal asymmetric neural stem cell division. Less is known about the expression and function of PROX1 in human brain tumors. First, we investigated expression of PROX1 using immunohistochemistry in 56 human astrocytic gliomas of different grades. PROX1 was found to be overexpressed in the majority of high grade tumors but not in grade I tumors or control tissue. Secondly, we addressed the prognostic value of PROX1 in a clinical cohort of patients with grade II gliomas (n=116). The number of PROX1 immunoreactive cells was used as a variable in survival analysis. Patients with tumors showing high PROX1 protein expression had poor outcome in the univariate analysis as well as in the multivariate analysis compared to those with low PROX1 expression. Thirdly, we found that PROX1 was expressed in glioma cell lines and we noted an apparent positive correlation between expression of PROX1 and the pluripotency factor SOX2. Depletion of SOX2 using siRNA inhibited glioma cell proliferation and reduced the levels of PROX1. Acute loss of PROX1 did not impair glioma cell proliferation in vitro. We conclude that PROX1 may constitute a useful tool for diagnosing and grading of astrocytic glioma distinguishing grade III and IV tumors from grade I and II tumors. PROX1 is also a novel predictor of survival for grade II gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5560. doi:1538-7445.AM2012-5560
Abstract 5563: Comparative studies of TIMP-1 immunohistochemistry, TIMP-1 FISH analysis and plasma TIMP-1 in glioblastoma patients
Authors: Aaberg-Jessen Charlotte , Jensen Stine S. , Nielsen Signe L. , Pedersen Christian B. , Brünner Nils , Kristensen Bjarne W.
Published: August 2012

Tissue inhibitor of metalloproteinases-1 (TIMP-1) has been associated with poor prognosis in various cancers as well as with resistance towards chemotherapy. In a previous study we found an association between a high immunohistochemical TIMP-1 protein level and a shorter survival for glioblastoma patients. The aim of the present study was to further evaluate TIMP-1 as a prognostic marker in gliomas by performing TIMP-1 fluorescence in situ hybridization (FISH) and by measuring TIMP-1 in plasma. TIMP-1 FISH analysis was performed on 33 glioblastoma biopsies in order to elucidate if the increased levels of TIMP-1 protein could be explained by gene amplification. Moreover, blood samples were collected from 43 patients (31 gliomas including 21 glioblastomas) prior to brain tumor surgery. The plasma TIMP-1 protein level was measured by ELISA. The results showed TIMP-1 FISH scores between 0.7 and 1.1, suggesting neither amplification nor loss of the TIMP-1 gene. The TIMP-1 level measured in plasma was not significantly higher than the TIMP-1 level measured in healthy matched controls. No correlation was identified between the immunohistochemical TIMP-1 levels and the FISH scores or the plasma TIMP-1 levels. In conclusion, TIMP-1 FISH scores or plasma TIMP-1 levels are without prognostic potential in gliomas. The present study suggests that the high immunohistochemical TIMP-1 protein levels in gliomas are not caused by TIMP-1 gene amplification. Moreover, TIMP-1 in plasma is low and not directly related to the tumor TIMP-1 level found by immunohistochemistry. The study suggests that TIMP-1 immunohistochemistry is the method of choice when using TIMP-1 as a biomarker in gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5563. doi:1538-7445.AM2012-5563
Abstract 5566: Elevated CXCR4 expression is clinically correlated with advanced stage neuroblastoma: examination of its role as a possible therapeutic target
Authors: Carlisle Alex J. , Pawal Bruce , Carlisle Rosalind , Jagannathan Jayanti , Martinez Daniel , Zhao Huaqing , Wang Qun , Cole Kristina , Lyttle Christopher A. , Nguyen Nhu , Maris John M.
Published: August 2012

ABSTRACT Purpose: Recent evidence suggests that expression of CXCR4 is associated with the growth and progression of neuroblastoma. The purpose of this study was to determine the value of CXCR4 as in independent predictive biomarker and evaluate its potential as a therapeutic target in neuroblastoma. Experimental Design: mRNA Expression profiling was performed on 101 diagnostic neuroblastoma primary tumor samples using Affymetrix oligonucleotide arrays. CXCR4 expression was stratified by risk and stage, and statistically significant differences in mRNA levels were determined. Protein expression profiling was performed by conducting immunohistochemical analysis of CXCR4 expression in 92 neuroblastoma tumors on a tissue microarray constructed at Children's Hospital of Philadelphia (CHOP); expression levels were stratified by stage and compared to determine significant correlations. To examine the role of CXCR4 in neuroblastoma progression, the selective CXCR4 inhibitor AMD3100 was used to observe the effect of CXCR4 inhibition on the growth of neuroblastoma cells in culture and in mice. Results: CXCR4 expression was significantly higher in stage 1 vs. ganglioneuroma tumors, as assessed at the protein level (P<0.0001), and in stage 4 tumors vs. stage 1 tumors, at both the mRNA and protein levels (P<0.01). Additionally, a comparison of CXCR4 transcriptional expression in patient tumors stratified by risk revealed greater CXCR4 expression in high-risk groups compared to low and intermediate groups (P<0.002); interestingly, CXCR4 protein expression was not associated with overall survival or progression-free survival (PFS). Treatment of neuroblastoma cells with AMD3100 inhibited growth of these cells in culture and in mouse xenografts. Conclusion: We conclude that increased levels of CXCR4 expression are associated with advanced tumor stage and a high risk population in neuroblastoma patients, demonstrating the clinical value of CXCR4 as an independent biomarker for risk classification and patient stratification; additionally, CXCR4 should be considered a novel and viable candidate for targeted therapeutic intervention in neuroblastoma patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5566. doi:1538-7445.AM2012-5566
Abstract 557: Fluorescence in situ hybridization versus qPCR as means to detect Merkel cell polyomavirus in Merkel cell carcinoma
Authors: Haugg Anke M. , Rennspiess Dorit , zur Hausen Axel , Speel Ernst-Jan M. , Cathomas Gieri , Becker Jürgen C. , Schrama David
Published: August 2012

Background: Merkel cell polyoma virus (MCPyV) is detected in 80% of Merkel cell carcinomas (MCC). The clonal integration and tumor specific mutations in the large T Antigen (LTAg) gene strongly implicate an oncogenic impact of the virus. To date the relationship between the viral presence and cancer induction, development or clinical prognosis is discussed controversial. Yet almost all studies are based on quantitative virus detection e.g. PCR or qPCR. Material and Method: In order to gain additional information about the quality of the viral presence on the single cell level we performed FISH analysis of formalin fixed and paraffin embedded (FFPE) MCCs (n= 62) on tissue micro arrays (TMA), determined the FISH pattern and correlated the results of the FISH analysis with the qPCR data. We grouped the MCCs according to different FISH evaluations and correlated them with the respective qPCR data on the basis of a determined cut-off. MCPyV FISH was established using the MKL-1 cell line which harbors integrated copies of MCPyV DNA. For MCPyV-qPCR the LT3 primer pair was used on whole tissue sections. Results: MCPyV-FISH on FFPE MKL-1 cells revealed punctate signals compatible with viral integration. The MCPyV FISH positive MCC cores (76%) mainly revealed two different signal patterns: a punctate pattern (85%) which correlated with a moderate relative viral presence and in some areas the punctate pattern was combined with a diffuse pattern (15%) indicating the episomal presence of the virus which is linked to viral replication. If a pattern mixture was seen this was associated with very high qPCR values. Comparing MCPyV FISH and qPCR data the results correlated highly with 83 % in the MCPyV positive evaluated group, whereas the negative group showed a concordance of 93 %. The mean of the qPCR values of all MCPyV positive cores differed significantly from the negative cores (p= 0.0076). The FISH signals were in some tumor areas from the same patient heterogenic in intensity, pattern and localization. Conclusion: While the qPCR using the LT3 primer pair was highly sensitive to detect MCPyV sequences in the respective MCC cases, the MCPyV FISH analysis highly concordantly revealed the quality of the viral presence, i.e. viral integration or episomal presence and morphological sublocalization in the tissues.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 557. doi:1538-7445.AM2012-557
Abstract 5573: Nuclear IGF-1R expression as a prognostic factor in cervical cancer patients
Authors: Tanioka Maki , Sudo Tamotsu , Obayashi Chiho , Matsumoto Koji , Fujiwara Kiyoshi , Negoro Shunichi , Minami Hironobu
Published: August 2012

Background: Insulin-like growth factor-1 receptor (IGF-1R) is an unusual transmembrane receptor in that both α and β subunits, regulated by IGF-1, translocate from the membrane to the nucleus and then bind to DNA, increasing gene transcription. Insulin receptor substrate-1 (IRS-1), a docking protein for IGF-1R, also traffics to the nucleus and then activates promoters of cell cycle progression genes. We studied the prognostic significance of nuclear and membranous IGF-1R and nuclear IRS-1 in cervical cancer, previously unreported. Methods: We retrospectively reviewed all patients (pts) with stage Ib2-IIb disease who received platinum-based neoadjuvant chemotherapy (NAC) at Hyogo Cancer Center between 2002 and 2009. Age, performance status, FIGO stage, histology, tumor size, pelvic lymph nodes on MRI, squamous cell carcinoma (SCC) antigen level, hemoglobin levels before NAC, chemotherapeutic agents and chemotherapy cycles were evaluated. Paraffin-embedded pretreatment tissue was stained immunohistochemically with an anti-IGF-1Rβ (Cell signaling, MA) or anti-IRS-1 (Millipore, MA) rabbit polyclonal antibody. IGF-1R and IRS-1 expressions were scored from 0 to 3 according to frequency and intensity. All slides were reviewed by a single pathologist. We analyzed prognostic factors associated with overall survival by a Cox proportional-hazards model. Results: A total of 128 pts (median age 57 years) were identified. The majority had stage II (61%) and SCC (69%). Median tumor size was 40 mm. Overall response rate to NAC was 59%, and median follow-up was 34 months. Nuclear and membranous IGF-1R and nuclear IRS-1 expressions were scored as 0/1-2/3 in 49/60/19 pts, 97/12/19 pts, and 65/41/22 pts, respectively. Multivariate analysis revealed that nuclear IGF-1R expression (HR, 2.6; P= .035 for score 1-3 vs. 0) and age <40 (HR, 2.6; P= .026) were independently associated with worse overall survival. Neither membranous IGF-1R nor IRS-1 expression had a prognostic impact on survival. Conclusions: Nuclear IGF-1R is commonly expressed in cervical cancer and is an independent prognostic factor, unlike membranous IGF-1R or nuclear IRS-1. Approaches that block the translocation of IGF-1R from the cell membrane to nucleus might hold promise for the treatment of cervical cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5573. doi:1538-7445.AM2012-5573
Abstract 5578: IGF-1R nuclear staining in tumor cells identifies sarcoma patients with a prolonged progression free survival after IGF-1R monoclonal antibody therapy
Authors: Asmane Irene , Watkin Emmanuel , Marec-Berard Perrine , Ray-Coquard Isabelle , Alberti Laurent , Cassier Philippe , Decouvelaere Anne-Valerie , Ranchere Dominique , Kurtz Jean-Emmanuel , Bergerat Jean-Pierre , Blay Jean-Yves
Published: August 2012

Rationale A small proportion of patients with sarcoma achieves prolonged progression free survival (PFS) with insulin growth factor type 1 receptor (IGF-1R) monoclonal antibody (Ab). A biomarker identifying in routine those patients beforehand would be useful to select patients. The aim of this study is to identify predictive factors for PFS in soft tissue sarcomas (STS), Ewing sarcoma (ES) and osteosarcoma patients receiving IGF-1R Ab. Methods The series of patients with unresecable or metastatic STS, ES, or osteosarcomas treated with IGF-1R Ab (R1507, IMC-A12, SCH 717454 and CP-751.871) in the Centre Leon Berard was studied. Tumor samples were analyzed by immunohistochemistry for expression of IGF-1R, IGFBP-3, Ki-67, HER1 and HER2. Results All tumor samples had a positive IGF-1R immunostaining on 60% to 100% of cells. IGFBP-3 immunostaining was observed in 12 (75%) samples with 5% to 100% of positive cells. IGF-1R immunostaining was nuclear (n=9, 56%), cytoplasmic (n=3, 19%), or nuclear + cytoplasmic (n=4, 25%). Neither IGFBP-3 expression, nor Ki67 was correlated to PFS or overall survival (OS). HER2 and HER1 stainings were positive in 0 and 2 samples respectively (both primary resistant to IGF-1R Ab therapy). Exclusive intra-nuclear immunostaining for IGF-1R was significantly associated with an increased PFS (p=0.009) and OS (p=0.04). Conclusion Exclusive nuclear localization of IGF-1R in sarcomas using routine immunohistochemistry was correlated to PFS and OS after IGF1R Ab treatment in this single center series. This observation is consistent with recent data reporting that nuclear localization of IGF-1R in tumor cells is a hallmark of pathway activation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5578. doi:1538-7445.AM2012-5578
Abstract 5579: Genomic approach to the prediction of chemotherapeutic sensitivity in osteosarcoma
Authors: Iwata Shintaro , Ohira Miki , Kamijo Takehiko , Yonemoto Tsukasa , Ishii Takeshi , Tatezaki Shin-ichiro , Nakagawara Akira
Published: August 2012

PURPOSE: Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. Despite of recent progress of combined therapy, about 30% of OS patients die because of tumor progression or relapse. It is generally accepted that sensitivity of neoadjuvant chemotherapy is the most important prognostic factor for OS, however, there have been few analyses documenting molecular markers that predicts the chemosensitivity of OS patients. The aim of this study is to identify possible genomic biomarkers for the prediction of chemosensitivity and prognosis for OS patients. METHODS: Pretherapeutic biopsy samples of 22 pediatric OS patients treated in Chiba cancer center who had homogeneous clinicopathological- and therapeutic- background were analyzed. In our study, patients were divided into 3 groups; group A includes 8 patients who demonstrated a good response for 1st regimen, group B includes 6 patients who were resistant to 1st regimen but showed good response to 2nd regimen, group C includes 8 patients who did not respond to neither 1st nor 2nd regimen. Genomic DNAs were prepared from each frozen tissue and used for aCGH analysis with Agilent 44k DNA microarray. RESULTS: According to the results of comparative analysis of CNV patterns in each patient, three chromosomal patterns strongly separated group A from B&C or C from A&B were identified. Gain of 5p12 was found in 7 out of 8 patients of group A, while only 1 out of 14 patients of group B and C (sensitivity 87.5%, specificity 92.9%). On the other hand, loss of 9q21 and amplification of 12q14 were specifically observed in group C, but not in A and B (sensitivity 100%, specificity 92.9%). The average of copy numbers within the region of 5p12 and 9q21.3 on each patient were calculated, and 2-dimensional plot of these two values clearly showed that each group formed distinct clusters. CONCLUSION: Our results indicated that copy number changes in OS biopsy samples could be good markers to determine suitable chemotherapeutic regimen for individual patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5579. doi:1538-7445.AM2012-5579
Abstract 5597: Modulating therapeutic effects of c-Src inhibitor via estrogen receptor and HER2 in breast cancer cell lines
Authors: Fan Ping , McDaniel Russell E. , Kim Helen R. , Jordan V Craig
Published: August 2012

Estrogen receptor (ER) and human epidermal growth factor receptor 2 (HER2) are two successful therapeutic targets in breast cancer. c-Src functions as an important adapter protein with ER and HER2, which validates it as an attractive target for the treatment of breast cancer. However, a very recent phase II clinical trial shows that a c-Src inhibitor, dasatinib, had limited single-agent activity in patients with HER2+ and/or hormone receptor (HR)+ advanced breast cancer (Mayer E et al. Clin Cancer Res 2011, Epub ahead of print). Therefore, a specific c-Src inhibitor, PP2, was utilized to block c-Src activity to identify targeted vulnerabilities affected by ER and HER2 in a panel of breast cancer cell lines (i.e. MCF-7, T47D, ZR-75-1, BT474, MCF-7:5C, MCF-7:2A, MDA-MB-231, Sk-Br-3, MCF-7/F, and T47D:C42). The antiproliferative effect of PP2 correlated with blocking c-Src mediated ERK/MAPK and/or PI3K/Akt growth pathways. The signaling pathways of c-Src were modulated by ER and HER2 in breast cancer cells. Inhibition of c-Src tyrosine kinase predominantly blocked ER negative breast cancer cell growth, particularly the triple (i.e. ER, PR, and HER2) negative cells. In contrast, ER negative Sk-Br-3 cells with highest HER2 phosphorylation were resistant to PP2, in which over-activated HER2 directly regulated growth pathways but not through c-Src. However, blocking c-Src recovered ER expression which made Sk-Br-3 cells regain responses to 4-hydroxytamoxifen. The majority of ER positive breast cancer cells were not sensitive to PP2 regardless of wild-type or endocrine resistant cell lines. Overall, c-Src mediates the essential role of growth pathways in ER negative breast cancer cells without HER2 over-activation. ER expression and HER2 over-activation are two important predictive biomarkers for the resistance to a c-Src inhibitor. These data provided an important therapeutic rationale for patient selection in clinical trials with c-Src inhibitors in triple negative breast cancer. Acknowledgements This work (VCJ) is supported by the SU2C (AACR) Grant (SU2C-AACR-DT0409).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5597. doi:1538-7445.AM2012-5597
Abstract 560: Development of an organotypic raft culture system to study the role of latent membrane protein 1 during Epstein-Barr virus replication
Authors: Geiser Vicki M. , Girard Julie , Godsil Mathias , Kahler Thomas , Rausch Emily , Rutledge Hayley
Published: August 2012

Epstein-Barr Virus (EBV) is a gamma herpesvirus that is prevalent worldwide and is an important cause of acute and chronic disease. Primary EBV infection which occurs after early childhood often results in acute infectious mononucleosis. Persistent latent EBV infection is a causative agent of anaplastic nasopharyngeal carcinoma, AIDS related lymphoma, posttransplant lymphoma, and Hodgkin's disease. Expression of Latent Membrane Protein 1 (LMP1) is common to most EBV related malignancies. LMP1 induced NF-κB activity is required for efficient transformation and survival of primary human B lymphocytes into lymphoblastoid cell lines. Increasing evidence suggest that LMP1 has an important role in the lytic cycle. Studies indicate that LMP1 and other latent genes are induced as part of the EBV lytic cascade in Burkitt's lymphomas. Our recent research indicates that LMP1-deleted virus replicates as efficiently as wildtype virus in HEK293 cells which suggests that the role of LMP1 during lytic replication is linked to changes in the cellular environment. LMP1 expression is associated with survival signals and alters expression of several differentiation markers, therefore a role for LMP1 during lytic replication in differentiating epithelial cells should be pursued. We have developed an organotypic raft culture system that will permit investigation of EBV replication in differentiating epithelial cells. Our lab is utilizing molecular, biochemical, and genetic approaches to elucidate the role of LMP1 during EBV replication in differentiating epithelial cells. We are investigating the function of LMP1 in viral replication, cell proliferation and differentiation, and viral-cellular signaling pathways. Based on these studies we will develop a model for the role of LMP1 during EBV replication in differentiating epithelial cells and its resulting pathogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 560. doi:1538-7445.AM2012-560
Abstract 5610: Activation of Ras/PI3K/ERK pathway induces c-Myc stabilization to upregulate argininosuccinate synthetase, leading to arginine deiminase resistance in melanoma cells
Authors: Tsai Wen-Bin
Published: August 2012

Arginine (Arg) depletion with pegylated recombinant arginine deiminase (ADI-PEG20), a novel strategy under clinical investigations for treating Arg-auxotrophic malignancies in humans, has produced promising results in several types of cancer, including malignant melanoma. These tumors are sensitive to ADI-PEG20 because they do not express argininosuccinate synthetase (AS), the rate-limiting enzyme for the de novo biosynthesis of Arg, and therefore require Arg from extracellular source for survival. ADI-PEG20 treatment, which catalyzes the conversion of Arg into citrulline and ammonia, results in Arg deprivation. Melanoma cells develop ADI-resistance because of induced AS upregulation. We previously demonstrated that induction of AS expression by ADI-PEG20 involves upregulation of transcription regulator c-Myc that positively regulates AS expression, but the underlying mechanism of c-Myc induction is not known. Here, we show that ADI-PEG20 activates Ras signaling and its downstream ERK and PI3K/Akt/GSK-3β kinase cascades. These cascades phosphorylate c-Myc, resulting in c-Myc stabilization by attenuating the ubiquitin-mediated protein degradation mechanism. Furthermore, we demonstrated that intervention of these signaling pathway using PI3K/Akt inhibitors suppressed c-Myc induction and enhance ADI-mediated cell killing. Because the Ras activation signaling is known to play important roles in tumor growth, our findings have important implications in Arg-deprivation and cancer cell metabolism and suggest that targeting the Ras signaling pathway may be beneficial in ADI-PEG20 anti-cancer therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5610. doi:1538-7445.AM2012-5610
Abstract 5611: A synthetic lethality combination with chloroquine and rapamycin on chemosensitization
Authors: Wang Yu-shan , Huang Yi-Chun , Fong Weng-Lam , Chi Mau-Shin , Chi Kwan-Hwa
Published: August 2012

Chemoresistance is the main obstacle of successful cancer therapy. Autophagy may either enhance cancer cell death or activate chemoresistance. Simultaneous pushing and blocking of autophagy may be a valid strategy for overcoming drug resistance and improving responses. We hypothesized that simultaneous forcing hepatoma cells synchronized in a pro-autophagic status by rapamycin while blocking the final autophagic pathway by chloroquine is an example of synthetic lethality in synergism. Co-administration of the chemotherapeutic agents with rapamycin and chloroquine in 3 hepatoma cell lines have all demonstrated synergistic effects including 5-fluorouracil, navelbine, taxotere or cisplatin. Increasing doses of rapamycin and chloroquine resulted in a significant decrease of ED50 of different chemodrugs. Althrough hepatoma cell lines expressed difference status of autophage during various chemodrug treatments, co-administration of rapamycin and chloroquine could synchronize them on the pro-autophagic status while preveting them ensuring to the final autophagic status in all cell line tested. The combination of rapamycin and chloroquine and chemotherapeutic agents resulted in increased apoptotic cell death compared to chemotherapeutic agents alone. All of the observed synergistic chemosensitization effect from combination of chloroquine and rapamycin is more obvious than either agent alone. In vivo experiments are underway. If these synergistic effects can be reproduced in vivo, the combination of rapamycin and chloroquine may provide a valid reversing resistance strategy in treatment refractory cancer patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5611. doi:1538-7445.AM2012-5611
Abstract 5612: The PI3K/Akt inhibitor LY294002 reverses BCRP/ABCG2-mediated drug resistance without BCRP translocation
Authors: Imai Yasuo , Yamagishi Hidetsugu , Ueda Yoshihiko
Published: August 2012

Cellular responses toward cytotoxic drugs are influenced by cross-talk between oncogenic signals and resistance mechanisms. Inhibition of the PI3K/Akt pathway is effective in sensitizing cancer cells of various organs, although the mechanisms have largely remained to be elucidated. Breast cancer resistance protein (BCRP)/ABCG2, a drug efflux pump, confers resistance to multiple anticancer agents such as SN-38 and topotecan. Previous studies reported that inhibition of the PI3K/Akt pathway, by gene knockout or PI3K inhibitors, modulated BCRP-mediated drug transport via BCRP translocation in hematopoietic stem cells, renal polarized cells, and glioma stem-like cells of mammals. In this paper, we tested reversing effects of PI3K inhibitors, LY294002 and wortmannin, on BCRP-mediated anticancer drug resistance of human cancer MCF-7 and A431 cells. LY294002 definitely reversed BCRP-mediated SN-38 and topotecan resistances but wortmannin did not. LY294002 treatment affected neither total nor cell surface BCRP levels by western blotting and flow cytometry but blocked BCRP-mediated topotecan efflux in a dose-dependent manner. Immunohistochemical analyses also demonstrated unchanged cellular BCRP distribution. BCRP overexpression in MCF-7 and A431 cells did not confer LY294002 resistance, suggesting that LY294002 would not be a transported substrate of BCRP. LY294002 is a derivative of quercetin, a member of flavonoid. Taken together, these results suggest that LY294002 inhibits BCRP-mediated drug transport not by BCRP translocation through the PI3K/Akt signal but putatively as a competitive inhibitor in a major subset of cancer cells. Due to its dual effects, LY294002 could be a lead compound for developing more effective and tolerable reagents for cancer treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5612. doi:1538-7445.AM2012-5612
Abstract 5615: Combination therapy for chronic myelogenous leukemia: Targeting the BCR-ABL coiled-coil domain and other pathways for synthetic lethality
Authors: Woessner David W. , Lim Carol S.
Published: August 2012

Single-agent tyrosine kinase inhibitor (TKI) therapy for chronic myeloid leukemia (CML) has been largely effective in keeping the disease at bay. However, resistance to therapy and persistence of a subset of leukemic cells demonstrate the necessity for multiple-agent therapy. We have developed a unique interfering peptide, CCmut2 (Dixon et al., JBC 2011) able to disrupt BCR-ABL dimers. This disruption of trans-auto phosphorylation exacts its activity via the coiled-coil domain in BCR, but leads to an overall similar effect as TKIs - induction of apoptosis and reduction in proliferation. Here we detail the results of this peptide (delivered as a gene and transcribed in vitro) in combination with small molecule or biologic agents. The secondary agents include: chloroquine which is used to inhibit lysosomal acidification and block cellular macroautophagy; zileuton, a specific inhibitor of arachidonate 5-lipoxygenase (Alox5) known to enhance the proliferative capacity of CML cells; and GO-201, an interfering peptide targeting MUC-1, a known oncogene expressed in CML cells. These agents in combination with CCmut2 were hypothesized to improve therapeutic potency and enhance leukemia-specific cytotoxicity. All assays were performed in BCR-ABL expressing K562 cells in vitro. Western blots confirming knockdown of the autophagy gene ATG7, Alox5, and MUC-1 using shRNA were carried out. The autophagy pathway was further validated by blotting for autophagic microtubule-associated protein light chain 3 (LC3) conversion from LC3-I to LC3-II following combination therapy with CCmut2 and chloroquine. Transformative ability of K562 cells was evaluated using a methylcellulose colony forming assay following co-transfection with CCmut2 and a shRNA. Next, the specific drugs targeting these pathways were also used in combination with the CCmut2 and assessed for apoptotic potential using caspase 3/7 activity and DNA segmentation. Proliferation was investigated using trypan blue exclusion and transformative ability was further assessed with multi-drug combinations. MUC-1 inhibition in combination with CCmut2 resulted in a potent combination in K562 cells. Other combinations hinted at an additive or synergistic enhancement of apoptosis and reduction in proliferation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5615. doi:1538-7445.AM2012-5615