bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract B15: Immunochemotherapy with 5-fluorouracil and interferon-enhances innate and adoptive immune responses against pancreatic cancer in vivo
Authors: Khallouf Hadeel , Serba Susanne , Jäger Dirk , Schmidt Jan 
Published: January 2011

Abstract
Pancreatic cancer has the poorest prognosis of all cancers urging the necessity of new therapeutic approaches. Recent clinical data show encouraging results for combining Interferon-α (IFN-α) immunotherapy with 5-Fluorouacil (5-FU) chemotherapy. However, the role of IFN-α in this immunochemotherapy scheme is still elusive. Here we investigate in vivo the relevance of different immune subpopulations in the anti-cancer immune response mediated by IFN-α in combination with 5-FU.
Luciferase-transfected Panc02 cells were implanted in the pancreas of C57BL/6 mice. Five days later, mice were treated with 5-FU alone, 5-FU + IFN-α or with a vehicle control. In parallel, CD4+ T cells, CD8+ T cells, and NK cells were depleted by neutralizing antibodies. Depletion of CD11c+ dendritic cells (DCs) was performed using transgenic CD11c.DTR mice. On day 21, tumor volume was measured and NK cells were isolated and enriched from spleens of mice then used as effectors against Panc02 cells in a standard chromium release assay. Tumors were frozen for further immunohistochemistry analysis to evaluate the immune cells infiltration. In parallel, we performed flow cytometry analysis to evaluate the effects of different treatments on the expression of MHC-I and NKG2D ligands (MULT-1 and Rae-1) on Panc02 cells.
Our data show that treatment with IFN-α + 5-FU has significantly decreased tumor volume in comparison with control or 5-FU treatments. This decrease in tumor volume was remarkably abolished by depleting CD8+ T cells or NK cells. Furthermore, NK cells isolated from spleens of IFN-α + 5-FU treated mice showed enhanced cytotoxicity towards Panc02 cells. Moreover, IFN-α + 5-FU treatment enhances the expression of MHC-I and NKG2D-ligand MULT-1 on Panc02 cells which could be the possible key for the enhanced recognition and per se killing by CD8+ T cells and NK cells respectively. To conclude, this study gives more insight about the mechanism of action of INF-α in combination with 5-FU and introduces CD8+ T cells and NK cells as main players in the observed anti-cancer immune response.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B15
Abstract B16: Expression of genes involved in PI3K/Akt signaling pathway in arsenic trioxide resistant clones derived from Jurkat cell line
Authors: Arkusz Joanna , Smok-Pieniazek Anna , Komorowska Magdalena , Stepnik Maciej 
Published: January 2011

Abstract
Arsenic trioxide (ATO) is a well-known carcinogen but also an effective cancer therapeutic drug for acute promyelocytic leukemia. Despite intensive research efforts, the molecular mechanisms underlying antileukemic effect of ATO as well as resistance to arsenic therapy appearing during treatment are not well understood. There are data showing that dysregulation of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway, a central integrating point of survival signals, may be the reason of apoptosis resistance in leukemic cells.
The aim of the present study was to assess expression of 84 genes involved in PI3K/Akt signaling pathway in Jurkat cells exposed to ATO at 1, 2.5 and 5 μM for 16 hrs as well as in ATO resistant clones derived from the cells. First screening of genes was performed using the Human PI3K-AKT Signaling Pathway RT2 ProfilerTM PCR Array (Superarray) and then selected genes were verified in a standard real-time PCR.
Results of our study showed that ATO induced more intensive changes in expression of PI3K/Akt pathway genes in ATO resistant clones than in Jurkat cells exposed to arsenic. Among genes differently regulated in both cell types were these playing a key role in regulating cell growth e.g. GRB10 (growth factor receptor-bound protein 10), FASLG (Fas ligand), CCND1 (cyclin D1). Interestingly, the level of FASLG mRNA was strongly down-regulated in ATO treated Jurkat cells and up-regulated in ATO resistant clones. Expression of CCND1 and GRB10 varied between resistant clones in a concentration-dependent manner. Moreover, concentration-dependent up-regulation of JUN (jun oncogene) was the most characteristic effect induced by ATO in Jurkat cells. Comparing to untreated Jurkat cells, we identified several genes differently regulated in ATO resistant clones (but not in Jurkat ATO exposed cells) that were mainly involved in anti-apoptotic pathways (e.g. IRS1 (Insulin receptor substrate 1), MAP2K1 (Mitogen-activated protein kinase kinase 1), MAPK3 (Mitogen-activated protein kinase 3), RPS6KA1 (Ribosomal protein S6 kinase, 90kDa, polypeptide 1), PRKCB (Protein kinase C, beta 1), PRKCZ (Protein kinase C, zeta) or Akt activation and growth regulation (e.g. PDK1 (Pyruvate dehydrogenase kinase, isozyme 1), MTOR (Mechanistic target of rapamycin), MTCP1 (Mature T-cell proliferation 1)).
In conclusion, our in vitro study identified several targets of ATO that may mediate cytotoxic effects in Jurkat cells. Moreover, the data suggest that the PI3K/Akt signaling pathway plays an important role in the ATO resistance in Jurkat leukemic cells. The work was supported by the Polish State Committee for Scientific Research (grant No PB 2806/B/PO1/2007/33).
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B16
Abstract B22: Beta-arrestin2-CARMA3 signaling axis plays critical roles in lysophosphatidic acid-induced ovarian cancer migration and invasion
Authors: Sun Jiyuan , Cai Jingli , Yumei Feng 
Published: January 2011

Abstract
Ovarian cancer is ranked as the 4th most common type of cancer among women in the United States. Lysophosphatidic acid (LPA) is the diagnostic marker of ovarian cancer. It is significantly elevated in 90% of ovarian cancer patients. Moreover, LPA receptors are aberrantly up-regulated in ovarian cancer patients. LPA is a type of G protein-coupled receptor ligand. In our previous studies, we have demonstrated that a signaling axis consisting of beta-arrestin 2 and CARMA3 (CARD and MAGUK domain-containing protein 3) is required in LPA-induced nuclear factor kappa B (NF-κB) activation in mouse embryonic fibroblast cells. NF-κB is an important transcription factor, which has been proven to mediate LPA-induced ovarian cancer migration and invasion. In this study, we further investigated whether the beta-arrestin2-CARMA3 signaling axis plays a significant role in LPA-induced ovarian cancer cell migration and invasion. Using beta-arrestin 2 and CARMA3 shRNA, we knockdowned their protein expression levels in ovarian cancer cell lines. Consistent with previous reports, we found that down-regulation of beta-arrestin 2 and CARMA3 abolished LPA-induced IKK activity and NF-κB activation in ovarian cancer cells. In addition, in vitro transwell migration and matrigel invasion assays demonstrated that beta-arrestin 2 and CARMA3 shRNA significantly impaired LPA-induced ovarian cancer cell motility and invasiveness. Together, our results provide the evidence that beta-arrestin 2 and CARMA3 serve as critical regulators in LPA-induced, NF-κB-mediated ovarian cancer migration and invasion. Therefore, we speculate that beta-arrestin 2 and CARMA3 may represent attractive therapeutic targets for ovarian cancer and many other types of malignancies.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B22
Abstract B24: Establishment of human primary non-small cell lung cancer xenograft models to test anticancer drugs
Authors: Liu Changnian , Li Wenwei , Li Bin , Liu Yong , Huang Wenpeng , Lou Boliang , Yang Hua 
Published: January 2011

Abstract
Background: Human xenograft tumor models established by transplantation of human tumor cell lines into immunodifficient mice have been routinely used for preclinical test of anticancer agents. But such tumor models have a relatively low transplantability and limited correlation with clinical findings. Recently, we have developed human non-small lung cancer (NSCLC) xenograft models by transplanting human fresh lung cancer tissues into nude mice, which have been employed for test of clinically used chemotherapeutical drugs and are under way for new drug chemosensitivity in vivo screening.
Methods: The fresh lung cancer samples were collected from local hospitals. The tumor fragments of 1–2 mm3 were subcutaneously implanted in the flanks of the Balb/c nude mice by trocar needle. Sixteen tumor fragments were grafted into four mice from one patient tumor tissue (passage 1). In the first passage, tumors derived from male patients were implanted into male mice, and tumors from women were inoculated into female mice. All therapeutic efficacy experiments used female mice. The drugs tested were clinically used drugs for NSCLC including cisplatin (4 mg/kg, q3d x 4), paclitaxel (15 mg/kg, q3d x 4), vinorelbine (2.5 mg/kg, q4d x 4), gemcitabine (120 mg/kg, q4d x 4), and the epidermal growth factor receptor (EGFR) inhibitor erlotinib (50 mg/kg/daily x 14, po).
Results: A total of 42 NSCLC samples were implanted, and 13 tumor models were established (tumor taking rate 31%) for the first passage. The tumor taking rates were higher in the second and third passages (80–100%). Cisplatin, paclitaxel, vinorelbine, and gemcitatbine produced tumor growth inhibition rates of 48–55% regardless of the EGFR mutation status. While erlotinib demonstrated a significant antitumor activity only in the tumors bearing EGFR, which were consistent with their clinical findings. The tumor xenografts' architecture, the cell and histopathological morphology from the three generations mirrored the original patient cancers.
Conclusions: These results suggest that human primary tumor xenograft models provide a unique renewable source of tumor material for test of novel anticancer agents. They may predict more relevant clinical response rates and higher correlation with clinical findings than use of xenograft models established from in vitro long-term cultured cancer cell lines, especially for test of target-oriented therapeutics in new drugs development programs.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B24
Abstract B27: The differential effect of all-trans retinoic acid and fenretinide in regulating differentiation and apoptosis, respectively of liver cancer cell
Authors: Wan Yu-Jui Yvonne , Bushue Nathan , Yang Hui 
Published: January 2011

Abstract
All-trans retinoic acid (RA) and its synthetic analog fenretinide [N-(4-hydroxyphenyl) retinamide] have very different effect on cancer cells. All-trans RA is a powerful differentiation agent and fenretinide is a reactive oxygen species (ROS) inducer that causes apoptosis of various types of cancer cell. It seems that both differentiation and apoptosis effect of these retinoids is mediated through retinoic acid receptor β (RARβ). In addition, both chemicals can induce the level of RARβ mRNA. However, our data showed that all-trans RA was a more effective RARβ inducer than fenretinide. The induction occurred within 30 min upon all-trans RA treatment vs. 3 hrs after fenetinide treatment in Huh7 cells. Moreover, both chemicals induced the expression level of LC3. However, the conversion of LC3-I to LC3-II, a hallmark of autophagy, was substantially higher in all-trans RA than fenretinide treated Huh7 cells. These data suggested the all-trans RA, but not fenretinide, was an effective autophagy inducer. In addition, the level of signaling adaptor p62, which is crucial for activation of death receptors and caspase 8-mediated apoptosis, was significantly lower in all-trans RA than fenrentinide treated Huh7 cells. Consequentially, all-trans RA-mediated autophagy might reduce ROS generation and inhibit apoptosis, whereas fenretinide caused ROS generation and cell death. The role of RARβ in differential regulation of LC3 expression and conversion as well as p62 expression remains to be determined. Taken together, the differential autophagy induction and p62 degradation might account for the different effect of all-trans RA and fenretinide in controlling the fate of liver cancer cells. (This research is supported by NIH grant CA53596.)
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B27
Abstract B28: Epigenetic contribution of Wnt antagonists SFRP1 and DKK1 as prognostic markers in colorectal cancer
Authors: Bapat Bharati , Rawson James B. , Mrkonjic Miralem , Green Roger C. , Gallinger Steve , Younghusband Banfield , McLaughlin John , Knight Julia A. 
Published: January 2011

Abstract
Background: Aberrant Wnt pathway activation is a vital carcinogenic event in colorectal cancer (CRC). DKK1 and SFRP1 encode extracellular inhibitors of canonical and canonical/non-canonical Wnt signaling, respectively, that are frequently silenced by promoter hypermethylation in CRC. Despite their known tumor-suppressive roles, few studies have systematically examined the prognostic/predictive significance of methylation in these genes in tumor development. Using a population-based genetic epidemiological approach, we investigated the methylation status of DKK1 and SFRP1 in a large cohort of primary CRCs and correlations to patient clinicopathological data.
Methods: As part of a Canadian interdisciplinary initiative to study the genetic and environmental determinants of CRC, we accrued a large number of primary colorectal carcinoma cases diagnosed in the province of Ontario, representative of a heterogeneous population (n = 558), and in the province of Newfoundland, representative of a founder population (n = 650). We examined the methylation status of DKK1 and SFRP1 gene promoters in colorectal tumors and matched normal colon tissues using MethyLight assay, a semi-quantitative methylation detection technique. We examined correlations between methylation levels and frequency, and a comprehensive array of patient clinicopathological features such as: age, sex, tumor stage, grade, tumor MSI subtype, and clinical outcome. Statistical analysis was performed using 2-tailed Fisher's exact test, SPSS v16.
Results: Respective DKK1 and SFRP1 methylation frequencies were similar in Ontario (13%, 95%) and Newfoundland (14%, 94%). Methylation was highly tumor specific. DKK1 methylation was a strong predictor of the microsatellite instability (MSI) tumor subtype in Ontario (OR=13.7 [7.8, 24.2], p &lt; 0.001) and in Newfoundland (OR=3.9 [2.8, 5.6] p &lt; 0.001). SFRP1 methylation was a predictor against MSI tumors in Ontario (OR=0.25 [0.15, 0.52], p &lt; 0.001).
Conclusion: Our study highlights the prognostic role of DKK1 and SFRP1 methylation related to CRC tumor subtype and suggests a novel distinction between the relative involvements of different Wnt pathways in microsatellite stable vs. unstable CRC.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B28
Abstract B29: Expression of BARD1 predicts outcome in colon cancer
Authors: Sporn Judith C. , Jung Barbara H. 
Published: January 2011

Abstract
Background: The BARD1/BRCA1 heterodimer has been implicated in several different pathways crucial for maintaining genomic integrity, DNA repair and cell survival. Complex functions have been ascribed to BARD1: It acts as a chaperone that recruits BRCA1 to the nucleus to form an ubiquitin E3 ligase complex involved in DNA repair and chromosomal stability. On the other hand, BARD1 may also promote cell survival by inhibiting BRCA1-dependent apoptosis through nuclear retention of BRCA1. Mutations in the BARD1 gene, as well as aberrant expression of BARD1 splice isoforms, have been associated with poor prognosis in breast and ovarian cancer. Here, we assess the occurrence and role of BARD1 splice isoforms in colon cancer formation.
Methods: Total RNA from primary human colon cancer samples and matched normal colon tissue was extracted and reverse transcribed. Gene specific PCR primers were designed to analyze the occurrence and pattern of BARD1 splice variants in colon cancer. Splice variants were subcloned and sequenced. Protein expression of BRCA1 and BARD1 in paraffin-embedded cancer tissues was detected by immunohistochemistry and correlated with clinicopathologic parameters.
Results: mRNA analysis of primary colon cancers confirmed a specific pattern of BARD1 splice isoforms distinct from normal controls. Certain isoforms appear to be tumor specific. Full length BARD1 is upregulated in non-metastatic cancer samples compared to matched normal tissue, which is in line with its anti-apoptotic function. Interestingly, full length BARD1 is dramatically downregulated in metastatic cancer tissue supporting its role as a tumor suppressor. Immunohistochemical analysis of colon cancer samples confirmed the loss of full length BARD1 in metastatic tissues on the protein level and revealed a significant correlation (p&lt;0.05) between expression levels of BARD1 and outcome, in which low expression of BARD1 is associated with a worse prognosis.
Conclusion: Our results suggest critical roles for the BRCA1 chaperone BARD1 in colon cancer formation. Alternative splice variants of BARD1 commonly occur in colon cancer tissue and expression of full-length BARD1 is differentially regulated along the cancer-metastasis axis affecting cancer outcome. We hypothesize that in colon cancer tissue, BRCA1 function critically depends on BARD1 expression.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B29
Abstract B3: A nested case control study of the polymorphic locus -202 in the promoter region of the IGFBP-3 gene and breast cancer risk
Authors: Macis Debora , Maisonneuve Patrick , Johansson Hariett , Aristarco Valentina , Guerrieri-Gonzaga Aliana , Rotmensz Nicole , Santillo Barbara , Gucciardo Giacomo , D'Aiuto Giuseppe , del Turco Marco Rosselli , Costa Alberto , Veronesi Umberto , Decensi Andrea , Bonanni Bernardo 
Published: January 2011

Abstract
Background: The Insulin-like Growth Factors (IGFs) system has been shown to have an important role in breast carcinogenesis due to its involvement in the regulation of cell proliferation, differentiation and apoptosis. Several studies have found an association between serum concentration of IGF-I and IGFBP-3 - its major binding protein - and breast cancer risk.
A single nucleotide polymorphism in the promoter region of the IGFBP-3 gene (rs2854744), located at position -202 from the transcription start site, results in reduced promoter activity and decreased IGFBP-3 levels. Some evidence of an association between this polymorphism and breast cancer risk has been reported.
Methods and results: We conducted a nested case-control study within the Italian Tamoxifen Chemoprevention Trial, a phase III double-blind placebo-controlled multicentric prevention trial involving 5,408 hysterectomized women aged 35–70 years, who had received either tamoxifen 20 mg/day or placebo for 5 years. After an initial median follow-up of 81.2 months, 79 subjects were diagnosed with breast cancer. Forty-six patients (cases) consented a blood withdrawal as well as 136 controls without breast cancer, randomly selected and matched for age (±2 years), randomization date (±6 months) and participating center.
We genotyped case and control subjects for IGFBP-3 -202A&gt;C with a Taqman real time PCR method for allelic discrimination (Applied Biosystems).
The IGFBP-3 -202A&gt;C genotype frequencies for AA, AC, CC were 46%, 43% and 11% respectively in breast cancer cases and 31%, 48%, 21% in controls. We found a significant trend for a higher frequency of A allele within breast cancer cases compared to unaffected control subjects (Ptrend=0.04). We observed an odds ratio of 0.35 (95% CI 0.12–1.02) for breast cancer in subjects with CC compared to AA genotype, while no further association was found after stratifying for age and treatment group.
Conclusions: We showed that in the analyzed population within the Italian Tamoxifen Chemoprevention Trial, the genotype frequencies for the IGFBP-3 -202 A allele were higher in breast cancer cases compared to unaffected control subjects at variance with other observations. These results suggest that variation at the polymorphic locus -202 in the IGFBP-3 gene might have an influence on breast cancer risk, although further studies are mandatory to clarify the direction of this association and the possible biological meaning and clinical implications.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B3
Abstract B30: Regulation of angiogenesis by the VEGF/miR-101/EZH2 axis
Authors: Smits Michiel , Mir Shahryar E. , Nilsson Jonas , van der Stoop Petra , Niers Anneke , Marquez Victor , Cloos Jacqueline , Breakefield Xandra , Krichevsky Anna , Noske David , Tannous Bakhos , Wurdinger Thomas 
Published: January 2011

Abstract
Rationale: Angiogenesis is a balanced process controlled by pro- and anti-angiogenic molecules of which the regulation is not yet fully understood. Besides classical gene regulation, miRNAs have emerged as post-transcriptional regulators of angiogenesis. In addition, epigenetic changes caused by histone modifying enzymes have emerged as modulators of angiogenesis as well. So far, the interplay between miRNA modulation and histone-modulating enzymes during angiogenesis is unclear.
Objective: To investigate the functions of miR-101 and of its target, the histone methyltransferase EZH2 in endothelial cells and angiogenesis.
Methods and Results: We show that the pro-angiogenic effects caused by VEGF-mediated down-regulation of miR-101 are partly mediated through reduced translational repression of the histone methyltransferase EZH2. In vitro, the sprouting and migratory properties of primary endothelial cell cultures were reduced by inhibiting EZH2 through up-regulation of miR-101, siRNA-mediated knockdown of EZH2, or treatment with 3-Deazaneplanocin A (DZNep), a small molecule inhibitor of EZH2 methyltransferase activity. In addition, we studied the effects of EZH2 inhibition on tumor angiogenesis in vivo. We found that systemic DZNep administration reduced the number of blood vessels in a subcutaneous glioblastoma mouse model, without showing adverse toxicities.
Conclusion: By determining a pro-angiogenic VEGF/miR-101/EZH2 axis in endothelial cells we provide evidence that there is a functional link between growth factor-mediated signaling, post-transcriptional silencing, and histone methylation in the angiogenesis process. Inhibition of EZH2 may prove therapeutical in diseases in which aberrant vascularization plays a role.
Citation Information: Clin Cancer Res 2010;16(7 Suppl):B30