bluuebunny/crossref_metadata_2025 huggingface dataset:
Numerical Predictions of Cavitating Flow around Model Scale Propellers by CFD and Advanced Model Calibration
Authors: Morgut Mitja , Nobile Enrico
Published: August 2012

The numerical predictions of the cavitating flow around two model scale propellers in uniform inflow are presented and discussed. The simulations are carried out using a commercial CFD solver. The homogeneous model is used and the influence of three widespread mass transfer models, on the accuracy of the numerical predictions, is evaluated. The mass transfer models in question share the common feature of employing empirical coefficients to adjust mass transfer rate from water to vapour and back, which can affect the stability and accuracy of the predictions. Thus, for a fair and congruent comparison, the empirical coefficients of the different mass transfer models are first properly calibrated using an optimization strategy. The numerical results obtained, with the three different calibrated mass transfer models, are very similar to each other for two selected model scale propellers. Nevertheless, a tendency to overestimate the cavity extension is observed, and consequently the thrust, in the most severe operational conditions, is not properly predicted.
An Analytic Study of the Reversal of Hartmann Flows by Rotating Magnetic Fields
Authors: Núñez Manuel
Published: August 2012

The effects of a background uniform rotating magnetic field acting in a conducting fluid with a parallel flow are studied analytically. The stationary version with a transversal magnetic field is well known as generating Hartmann boundary layers. The Lorentz force includes now one term depending on the rotation speed and the distance to the boundary wall. As one intuitively expects, the rotation of magnetic field lines pushes backwards or forwards the flow. One consequence is that near the wall the flow will eventually reverse its direction, provided the rate of rotation and/or the magnetic field are large enough. The configuration could also describe a fixed magnetic field and a rotating flow.
Silent Intravascular Lymphoma Initially Manifesting as a Unilateral Adrenal Incidentaloma
Authors: Takahashi Yoshiko , Iida Keiji , Hino Yasuhisa , Ohara Takeshi , Kurahashi Toshifumi , Tashiro Takashi , Chihara Kazuo
Published: August 2012

Intravascular large B-cell lymphoma (IVLBCL) is a rare subtype of malignant lymphoma. Although the involvement of adrenal glands in IVLBCL is often observed, primary adrenal IVLBCL is rare. Most reported cases of adrenal IVLBCL showed bilateral lesions resulting in rapidly progressive adrenal failure and poor prognosis. Here, we report a case of slowly progressive primary adrenal IVLBCL manifesting initially with unilateral adrenal incidentaloma. This case is a silent IVLBCL and shows that the enlargement of both adrenal glands can be followed.
Migraine-Asthma Comorbidity and Risk of Hypertensive Disorders of Pregnancy
Authors: Czerwinski Stefanie , Gollero Jolana , Qiu Chunfang , Sorensen Tanya K. , Williams Michelle A.
Published: August 2012

Background. To evaluate the association of migraine and asthma and to estimate the risk of hypertensive disorders of pregnancy in relation to maternal comorbid migraine and asthma.Methods. Reproductive age women (N=3.731) were interviewed during early pregnancy. At the time of interview, we ascertained participants’ migraine and asthma status. From medical records, we collected information to allow the diagnosis of pregnancy-induced hypertension (PIH) and preeclampsia. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using logistic regression procedures.Results. After adjusting for confounders, migraineurs had 1.38-fold increased odds of asthma as compared with nonmigraineurs (95% CI 1.09–1.38). The odds of hypertensive disorders of pregnancy were highest among women with comorbid migraine-asthma. The ORs for PIH preeclampsia and the two disorders combined were 2.53 (95% CI 1.39–4.61), 3.53 (95% CI 1.51–8.24), and 2.64 (95% CI 1.56–4.47), respectively, for women with comorbid migraine-asthma as compared with those who had neither disorder.Conclusion. These findings confirm prior reports and extend the literature by documenting particularly high odds of pregnancy-induced hypertension and preeclampsia among women with comorbid migraine-asthma. Increased knowledge about the prevalence and sequelae of comorbidities during pregnancy may lead to improved symptom management and perinatal outcomes.
The Dual of Generalized Fuzzy Subspaces
Authors: Chen Wenjuan , Guan Yanyong
Published: August 2012

We define the dual of generalized fuzzy subspaces first. These concepts generalize the dual of fuzzy subspaces. And then we investigate the double dual of generalized fuzzy subspaces.
Role ofp16INK4Ain Replicative Senescence and DNA Damage-Induced Premature Senescence in p53-Deficient Human Cells
Authors: Mirzayans Razmik , Andrais Bonnie , Hansen Gavin , Murray David
Published: August 2012

Thep16INK4A(hereafter p16) tumor suppressor is encoded by theINK4A/ARFlocus which is among the most commonly dysregulated sequences in human cancer. By inhibiting cyclin-dependent kinases, p16 activates the G1-S checkpoint, and this response is often considered to be critical for establishing a senescence-like growth arrest. Not all studies support a universal role for p16 in senescence. Single-cell analysis of noncancerous human fibroblast cultures undergoing senescence as a function of culture age (replicative senescence) has revealed that p16 is not expressed in the majority (>90%) of cells that exhibit features of senescence (e.g., flattened and enlarged morphology coupled with senescence-associatedβ-galactosidase expression), ruling out a requirement for p16 in this process. In addition, ionizing radiation triggers premature senescence in human cancer cell lines that do not express p16. These observations are made with cells that express wild-type p53, a key mediator of the DNA damage response. In this paper, we examine the growing evidence suggesting a negative regulatory relationship between p16 and p53 and discuss recent reports that implicate a role for p16 in replicative senescence and ionizing radiation-induced premature senescence in human cells that lack wild-type p53 function.
Inhibition of Corrosion of Carbon Steel in Well Water by DL‐Phenylalanine‐Zn2+ System
Authors: Raja A. Sahaya , Rajendran S. , Satyabama P.
Published: August 2012

The environmental friendly inhibitor system DL‐phenylalanine‐Zn2+ has been investigated by weight loss method. A synergistic effect exists between DL‐phenylalanine and Zn2+ system. The formulation consisting of 150 ppm of DL‐phenylalanine and 5 ppm of Zn2+ offers good inhibition efficiency of 90%. Polarization study reveals that this formulation functions as a anodic inhibitor. AC impedance spectra reveal that a protective film is formed on the metal surface. The surface morphology has been analysed by SEM and EDAX. A suitable mechanism of corrosion inhibition is proposed based on the results obtained from weight loss study and electrochemical studies.
Abstract 1113: Role of the microRNA-23∼27∼24 clusters in osteosarcoma
Authors: Francis Princy , Moon So Young , Bilke Sven , Zhu Yuelin J. , Walker Robert L. , Pineda Marbin , Li Xiao Ling , Abaan Ogan , Knight Jeffrey , Davis Sean , Turner Jaleisa , Wang Hui , Bruhn Laurakay , Lal Ashish , Meltzer Paul S.
Published: August 2012

Osteosarcomas are rare but aggressive tumors that predominantly affect adolescents. These neoplasms of mesenchymal origin are biologically heterogeneous and produce immature osteoid or bone. The pathogenesis of osteosarcomas is poorly understood although defective osteogenic differentiation seems to play a key role. In order to elucidate the underlying molecular mechanisms, 16 osteosarcoma cell lines, 2 osteoblast cell lines and 45 osteosarcoma tumors were studied using a combination of high-throughput techniques such as microRNA (miRNA) and gene expression profiling, miRNA sequencing and array comparative genomic hybridization. Overexpression of the miRNA clusters miR-23a∼27a∼24-2 (19p13.13) and miR-23b∼27b∼24-1 (9q22.32) was identified in a subset of the osteosarcoma tumors and cell lines. Furthermore, quantitative RT-PCR revealed that the miR-23a and miR-23b clusters were downregulated during osteoblast differentiation of the osteosarcoma cell line HOS and overexpression of these miRNAs (by transient transfection of miRNA mimics) delayed osteoblast differentiation and matrix mineralization evaluated by alkaline phosphatase and Alizarin Red staining. Target gene identification experiments were then performed for each of the members of the miR-23a and miR-23b clusters. Gain of function (using miRNA mimics for exogenous expression) and loss of function (using antisense miRNAs to inhibit endogenous expression) experiments followed by downstream gene expression analysis identified several putative targets in osteosarcoma cell lines HOS and HuO3N1. The gene lists were highly enriched in targets (including differentiation genes) identified by TargetScan and other target prediction software. These results suggest that deregulation of the miR-23∼27∼24 clusters may play a role in defective osteogenic differentiation and osteosarcoma.

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 1113. doi:1538-7445.AM2012-1113
Abstract 1116: Evaluation of mitochondrial DNA G10398A polymorphism in Iranian breast cancer patients
Authors: Ghaffarpour Massoud , Hosseini Zahra Mostakhdemin , Kamali Fatemeh , Fereidooni Forouzandeh , Moazami Nasrin , Houshmand Massoud
Published: August 2012

Introduction: The role of mtDNA alteration is already recognized as being in carcinogenesis, tumor progression and prognosis in all type of cancers. mtDNA alteration in complex I have been reported in different cancers. An A to G polymorphism at nucleotide position 10398 caused a nonconservative amino acid substitution from threonine to alanine within the NADH dehydrogenase (ND3) subunit of Complex I. The mitochondrial DNA 10398 polymorphism G10398A within ND3 subunit of Complex I has been suggested to alter the mitochondrial subunit of electron transfer chain and has been associated with several neurodegenerative disorders and cancers. It has been also shown that mtDNA alteration G10398A within ND3 subunit of Complex I in breast cancer patients correlated with increased breast cancer risks in the Indian and African-American populations however, the 10398A allele was not increase breast cancer risk for Caucasian women. Interestingly, G10398A in Chinese population were shown correlated with breast cancer metastasis. The aim of this study was on the association of mtDNA G10398 A polymorphism in Iranian breast cancer patients. Materials and Methods: Patients and tissue specimens: 27 paired Fresh tumor and adjacent normal samples were obtained from consecutive patients with BC (age range, 33-74years; with median age of 49.16 years) who underwent surgery for a partial and or whole mama mastroctomy between October 2007 and Nov 2009 from IRANIAN National Tumor Bank, National Cancer Institute, Imam Khomeini Hospital Complexes, Medical Tehran University, Tehran, Iran. Histopathological examinations were performed by two expert's pathologists. Mutational analysis of mtDNA G10398A in tumor samples: fresh tumors and their adjacent were extracted for genomic DNA using the QIAamp Mini Kit (Qiagen in accordance with the manufactures instructions. We searched for 10398 Polymorphism in within ND3 subunit of Complex I by means of PCR sequencing. Result and conclusion:patient had histopathological grade I, II, and III with 66.6%, 25.9% and 7.4% respectively. The frequency of mtDNA G10398A polymorphism 44.440%(12 of 27).The Iranian women with G10398A variant showed a significantly increased risk of breast cancer (p< 0.05). Derived from these findings, mtDNA G10398A polymorphism may be a potential risk marker of breast cancerk in Iranian population.

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 1116. doi:1538-7445.AM2012-1116
Abstract 113: Effect of targeting Aurora kinase A in human oral squamous cell carcinoma cells
Authors: Tanaka Hiroshi , Nakashiro Koh-ichi , Oka Ryota , Hamakawa Hiroyuki
Published: August 2012

Oncogene addiction has provided therapeutic opportunities in many human malignancies, but molecular targeted therapy for oral squamous cell carcinoma (OSCC) is not available. In this study, we identified an appropriate molecule for treatment of patients with OSCC. We determined the gene expression profiles in 10 human OSCC cell lines and a non-neoplastic keratinocyte cell line by microarray analysis, and then identified Aurora kinase A (AURKA) as a cancer-related gene which was commonly overexpressed in OSCC cells. We also confirmed the overexpression of AURKA mRNA in OSCC tissues and compared to normal oral mucosa tissues by quantitative RT-PCR. AURKA plays an essential role in centrosome duplication, mitoic entry, and spindle assembly. Overexpression of AURKA in human malignancies is associated with gene amplification, genetic instability, poor histological differentiation, and poor prognosis. To determine if the AURKA is a plausible therapeutic target, we investigated the effect of small interfering RNAs specific for AURKA(siAURKA) and an AURKA selective inhibitor MLN8237 on the growth of human OSCC cells in vitro and in vivo. Knockdown of AURKA by RNA interference (RNAi) markedly inhibited the growth of OSCC cells and observed the change of colony formation and cell size looked like cell cycle arrest. We examined the RNAi effect of siAURKA in OSCC cells by Western blotting. Compared with non-target siRNA, the protein expression level of AURKA was almost completely suppressed. MLN8237 also reduced the cell growth, but showed relatively resistant to cells expressing AURKA at high levels. Transfection of siAURKA inhibited the growth of OSCC cells more potent than treatment with MLN8237 in vitro. Furthermore, we confirmed the effect of siAURKA and MLN8237 on the growth of human OSCC tumors which had been subcutaneously xenografted in athymic nude mice. Atelocollagen-siAURKA (8 nmol) complexes were injected into the tail vein every 3 days and MLN8237 (30 mg/kg) were received orally for 14 consecutive days. We found that siAURKA and MLN8237 significantly reduced the size of subcutaneously xenografted OSSC tumors. Interestingly, the combination of siAURKA and MLN8237 induced remarkable anti-tumor activity compared with single agent groups. These results suggest that AURKA functions as a critical gene for supporting the growth of human OSCC cells and targeting AURKA may be a useful therapeutic strategy for OSCC.

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 113. doi:1538-7445.AM2012-113
Abstract 1147: Molecular characterization of PI3K pathway alterations in non-small cell lung carcinoma
Authors: Huw Ling-Yuh , Spoerke Jill , O'Brien Carol , Koeppen Hartmut , Fridlyand Jane , Amler Lukas , Hampton Garret , Shames David , Lackner Mark , Pandita Ajay
Published: August 2012

Lung cancer is the leading cause of cancer deaths worldwide. Several oncogenes and tumor suppressor genes have been implicated in lung carcinogenesis. Among them, PIK3CA pathway activation is one of the key pathways that link cell surface receptor kinases to effectors for tumor growth, metabolism and survival. To support predictive diagnostic biomarker efforts for the selective PI3K inhibitor GDC-0941, we conducted detailed molecular characterization of NSCLC tumor samples. We evaluated the key oncogenic mutations (KRAS, EGFR, PIK3CA, BRAF and NRAS), gene expression, protein expression (PTEN) and gene copy number alterations in genes in the PI3K pathway in NSCLC tumors. We found that oncogenic mutations and copy number alternations were associated with histological subtype. Of note, mutations in KRAS and EGFR were found with high frequency (30%) in adenocarcinoma, as was loss of the tumor suppressor LKB1. On the other hand, amplification of PI3K was found in the squamous subtype, as were rare mutations in PI3K. The tumor suppressor PTEN showed a high frequency of reduced expression in both squamous and adenocarcinomas, though total loss of PTEN was more common in squamous carcinomas. Our studies suggest that the PI3K pathway is an attractive therapeutic target in NSCLC, and that distinct biomarker strategies may be required for development in squamous and non-squamous patient populations.

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 1147. doi:1538-7445.AM2012-1147
Abstract 1167: The expression of urokinase plasminogen activator (uPA) system genes in prostate carcinoma is related to clinicopathological parameters
Authors: Al-Janabi Omar , Wach Sven , Nolte Elke Nolte , Wullich Bernd , Taubert Helge , Füssel Susanne , Lohse Andrea , Kotzsch Matthias , Wieland Wolf , Stoehr Robert , Hartmann Arndt
Published: August 2012

Background: Prostate cancer (PCa) is a leading cause of tumor mortality. Members of the urokinase plasminogen activator (uPA) system play an important role in tumorigenesis in many human malignancies. We sought to characterize the mRNA expression of uPA members in primary PCa samples and to correlate this expression to clinicopathological parameters. Material and methods: We investigated the mRNA expression level of three members of the uPA system (uPA, PAI-1 and uPAR) using 26 paired samples of PCa and adjacent non-malignant tissue. Absolute quantification of gene expression was performed by quantitative PCR and subsequent standardization on the reference gene TBP. Gene expression measures were further correlated to clinicopathological characteristics as TNM classification, Gleason sum, and tumor grade. Results: Although we did not detect significant differences in expression of uPA system genes between tumor and matched nonmalingnant tissue in our patient cohort, we identified a positive correlation of all three uPA family members (p<0.01; bivariate correlation). When stratifying PAI-1 expression according to the median mRNA level we discovered correlations to defined tumor characteristics. Tumors with PAI-1 expression above median displayed a higher Gleason sum (8 or 9) than tumors expressing less-than-median PAI-1 (p=0.03, Fisher's exact test). Likewise, an elevated PAI-1 expression was correlated with higher tumor grade (p=0.01; Fisher exact test) and tumor stage (p=0.046; Fisher exact test). Conclusion: Our results demonstrate that mRNA expression of PAI-1 as a member of the uPA family correlates with characteristic pathological parameters of PCa.

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 1167. doi:1538-7445.AM2012-1167
Abstract 1176: Bypassing the dominant-negative effect of mutant p53 in cancer cells
Authors: Okal Abood , Mossalam Mohanad , Matissek Karina J. , Lim Carol S.
Published: August 2012

The tumor suppressor p53 is mutated in more than 50% of all cancers. The transcriptional activity of p53 depends on localization to the nucleus and formation of p53 tetramers. Use of wild-type (wt) p53 for gene therapy is limited due to the dominant-negative effect of mutant (mut) p53 over wt p53. Mut p53 will hetero-oligomerize with wt p53 via the tetramerization domain (TD) found in p53, deactivating the tumor suppressor function of wt p53. In order to use p53 for gene therapy, an alternative oligomerization domain for p53 was investigated. The coiled-coil (CC) domain from Bcr (breakpoint cluster region), similar to the native TD of p53, can form an antiparallel dimer of dimers. The TD from p53 was then substituted with this CC, and the activity of this new p53 (p53-CC) was tested. We evaluated the transcriptional activity of p53-CC compared to wt p53 in a dual luciferase reporter gene assay. To demonstrate the ability of our p53-CC to bypass the dominant-negative effect prompted by hetero-oligomers, the reporter gene assay was carried in human breast adenocarcinoma (MCF-7) and human breast ductal carcinoma (T47D) cell lines. MCF-7 cells contain wt p53 population that is mislocalized to the cytoplasm, whereas, T47D cells contain mutant p53. Indeed, data from the reporter gene experiment shows a reduction in wt p53 activity introduced to T47D cells compared to MCF-7 cells, while the activity of p53-CC remains consistent regardless of the presence of an endogenous mut p53 population. We also tested the biological activity of p53-CC and its ability to induce apoptosis using 7-AAD and Annexin-V assays (late stage apoptosis). In addition, Caspase-3/7 assay (mid-stage apoptosis) was performed to further validate a p53-dependent apoptotic pathway. Data obtained from the aforementioned experiments suggest that p53-CC has a very similar biological activity to wt p53. Furthermore, we examined the gene expression profile of p53-CC using wt p53 as a reference. The assay was performed using real-time PCR array technology to analyze the expression of 84 genes related to p53-mediated signal transduction. Two negative controls were used; the first is a p53 construct that lacks its tetramerization domain (p53ΔTDC) and should not possess any transcriptional activity, and the second is the CC by itself. Indeed, p53-CC shows comparable gene expression profile to wt p53 and has the ability to transactivate the various p53 target genes. In summary, p53-CC retains p53 activity and excludes hetero-oligomer formation, overcoming a major barrier in using p53 for cancer gene 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 1176. doi:1538-7445.AM2012-1176
Abstract 1181: P73 regulates the expression of KAI1, metastasis suppressor gene
Authors: Lee Ji Hee , Bae Woo Kyun , eul Hwang Jun , Cho Sang Hee , Chung Ik Joo
Published: August 2012

KAI1, a metastasis suppressor gene of various cancers, has been known to inhibit tumor metastasis without affecting primary tumor growth. Identification of transcription factor regulating KAI1 is important for understanding tumor metastasis mechanism. P73 binding site within the KAI1 promoter were characterized by luciferase assay using reporter plasmids containing serial deletion promoter regions of KAI1 gene and chromatin-IP assay. To confirm whether the over-expressed KAI1 gene regulated by p73 in the transcriptional level was active, cell motility was analysed by using an adhesion and invasion assay. Here, we first identified that KAI1 is transcriptionally regulated by p73, a p53 family member, in various cancer cell lines. In p53-deficient cell lines, p73 had highest promoter binding activity in the KAI1 promoter regions between −903 and −846 bp, known as a p53-binding site. Human colon cancer cells stably overexpressing p73 showed that in vitro extracellular matrix adhesion and invasive potential was decreased compared with parental cells. These data explain that p73 regulate the KAI1 gene expression in the transcription level by binding on the specific region of the KAI1 promoter. In addition, it could be suggested that decreased invasion suppressive effect of KAI1 in invasive tumor and tumor cell lines caused by weak expression of KAI1 could be restored by increasing the p73 level. P73 might contribute the metastasis suppressive effect through upregulating KAI1 gene expression in invasive tumor and tumor cell lines. Taken together, our present results suggest that p73 contributes to KAI1 promoter activity in cells and plays an important role in the repression of tumor metastasis function via regulation of KAI1 expression in invasive tumor and tumor cell lines.

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 1181. doi:1538-7445.AM2012-1181
Abstract 131: Hypomethylation of Alu repeat sequences permits activation of microRNAs in human colorectal cancer metastasis
Authors: Hur Keun , Boland Clement Richard , Goel Ajay
Published: August 2012

Background: Global hypomethylation of cytosines within CpG dinucleotides has been reported in various human cancers. Repetitive elements dispersed throughout the human genome, including Alu and LINE1 sequences, have been used as global surrogate markers for estimating global DNA methylation levels. Recent evidence indicates that certain human miRNAs are located downstream of Alu sequences, and these non-coding RNAs can be transcribed by the recruitment of RNA polymerase III to the promoter region of Alu repeats. Primary colorectal cancers (CRC) demonstrate frequent global Alu hypomethylation; however, it is not clear whether hypomethylation of these repeat elements may be associated with a metastatic phenotype. Aim: This study aimed to determine the role of Alu hypomethylation in the development of CRC metastasis. Materials and Methods: We analyzed a panel of CRC cells, as well as matched pairs of primary CRCs (PC) and corresponding liver metastasis (LM) tissues from 59 CRC patients. Alu methylation levels were measured by quantitative bisulfite pyrosequencing. In addition, we determined the expression status of miR-520c and miR-373, which are located immediately downstream of Alu repeats. Results: All CRC cell lines demonstrated Alu hypomethylation, which was further exacerbated by treatment with the DNA demethylating agent, 5-azacytidine. In the clinical specimens, the levels of Alu methylation were significantly lower in LM compared to the matched PC (mean methylation value; PC = 83% vs. LM = 80%; P < 0.001). Moreover, methylation analysis of surrounding normal liver tissues revealed higher levels of Alu methylation (mean methylation value; 85%) compared to both PC and LM. The expression of both miR-520c and miR-373, showed significantly different expression between PC and LM, which was mediated by the hypomethylation of Alu-associated promoter region. Conclusion: Our results provide novel evidence for increased Alu hypomethylation in liver metastasis tissues from patients with CRC. We speculate that hypomethylation of Alu repeat elements permits activation of metastasis-related miRNAs, which in turn may facilitate a more aggressive malignant phenotype in these advanced stage cancers.

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 131. doi:1538-7445.AM2012-131
Abstract 1326: Magnetization transfer MRI in pancreatic cancer xenograft models
Authors: Li Weiguo , Zhang Zhuoli , Nicolai Jodi , Yang Guang-Yu , Omary Reed , Larson Andrew
Published: August 2012

Tumor desmoplasia is a common hallmark of pancreatic ductal adenocarcinoma (PDAC), the 4th leading cause of cancer-related mortality. Recent studies clearly indicate that desmoplasia actively facilitates disease progression, metastasis, and drug resistance. Precise quantification of tumor fibrosis levels in PDAC may be critically important for patient staging and for the prediction or monitoring of responses to therapy. The current gold-standard for characterizing desmoplasia_histologic analysis of biopsy samples_is invasive and cannot avoid misrepresentation by reason of sampling error. Therefore, new methods to permit noninvasive quantification of fibrosis levels in PDAC would be particularly useful. Magnetization transfer (MT) MRI can be used to non-invasively probe dynamic physical processes involving the exchange of magnetization between sub-populations of free water protons and those water protons bound to tissue macromolecules. MT-MRI studies have demonstrated that MT contrast (MTC) can be highly sensitive to tissue collagen concentration, the primary component of desmoplasia. Thus, we hypothesized that desmoplasia-associated fibrosis levels in PDAC may be directly reflected in MT-MRI magnetization transfer ratio (MTR) measurements. The purpose of this study was to compare non-invasive MTR measurements to invasive histologic assessments of tumor fibrosis-levels in three mouse xenograft models of PDAC. In this study, MT MRI measurements were performed in 3 PDAC mouse xenograft models. For each of 28 PDAC xenografts, MTR were calculated and compared to histologic fibrosis levels from reference standard trichrome staining. MTR was found to be significantly higher in tumors grown using BxPC-3 cell line (39.4 ± 5.1, mean ± SD) compared to the MTR for the tumors grown from Panc-1 (32.4 ± 2.8) and Capan-1 (27.3 ± 2.9) cell lines (P < 0.05 for each comparison). Histologic measurements showed a similar trend with BxPC-3 tumors demonstrating significantly higher fibrosis levels (percentage of fibrotic tissue area, 6.48 ± 2.59) when compared to Panc-1 (3.54 ± 2.18) and Capan-1 (2.07 ± 1.60) tumors. MTR measurements were well correlated to quantitative fibrosis levels (r = 0.69, P = 0.01). Our results indicated that MTR measurements offer the potential to serve as a valuable in vivo biomarker of desmoplasia in PDAC.

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 1326. doi:1538-7445.AM2012-1326
Abstract 1359: Establishment and characterization of mouse malignant rhabdoid tumor cell lines
Authors: Kuwahara Yasumichi , Mora-Blanco E Lorena , Rogers Arlin , Fletcher Christopher DM , Roberts Charles WM , Weissman Bernard E. , Weissman Bernard E.
Published: August 2012

Malignant rhabdoid tumors (MRTs) are rare, aggressive cancers occuring in young children primarily through inactivation of the SNF5 tumor suppressor gene. Espetially, rhabdoid tumor arised from brain is usually called AT/RT. We and others have demonstrated that mice heterozygous for a Snf5 null allele develop MRTs with partial penetrance. We have also shown that Snf5+/− mice that lacked expression of the pRb family, due to TgT121 transgene expression, developed MRTs with increased penetrance and decreased latency. In this current study, we observed that the genetic background altered MRT development in Snf5+/− and TgT121;Snf5+/− mice, resulting in increased latency and the appearance of significant frequency of brain tumors. We also report the establishment of the first mouse MRT cell lines that recapitulate many features of their human counterparts including lack of genomic instability and expression of smooth muscle and neuronal differentiation markers. The cell lines also form aggressive tumors upon inoculation into immunocompromised mice that resemble the tumors of origin. Our studies provide further insight into the genetic influences on MRT development as well as provide valuable new cell culture and genetically engineered mouse models for the study of AT/RT etiology.

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 1359. doi:1538-7445.AM2012-1359
Abstract 136: Suppression of the uPA/uPAR system causes the overexpression of miR124 and suppresses stemness in glioma stem cells
Authors: Asuthkar Swapna , Gorantla Bharathi , Rao Jasti S. , Gondi Christopher S.
Published: August 2012

The invasiveness and destructiveness of malignant neoplasms in the central nervous system are of great clinical importance. Higher-grade tumors, such as glioblastomas, are associated with poor prognosis, and patients’ average survival is only 8 to 12 months after chemotherapy and/or radiotherapy. This poor prognosis reflects the resistance of tumor cells to radiation and cytotoxic agents as well as the difficulty in achieving total tumor resection. Highly infiltrative gliomas are known to overexpress both uPA and uPAR. MicroRNAs (miRNAs) are essential post-transcriptional regulators known to determine cell identity and fate and are also known to be involved not only in development and differentiation but also in glioma progression. Acquisition of EMT has also recently been linked to stem cell phenotypes which is mediated by microRNAs. However, the molecular mechanism underlying EMT regulation still remains elusive. In the present study, we used glioma initiating cells (GICs), which show stem cell-like character, and determined whether their stemness can be suppressed by targeting the uPA/uPAR system. We raised glioma GICs from U87MG and 4910 glioma xenograft cells and observed that GICs expressed 2- to 3-fold increased levels of uPA and uPAR when compared to non-GIC. Further, the simultaneous downregulation of both uPA and uPAR suppressed GIC from establishing intracranial tumors in nude mice. Further, to understand whether this suppression of GIC involved microRNAs, we profiled miR expression in U87 cells and observed that miR124 and miR200a were significantly overexpressed. Using miRanda analysis, we determined that miR124 targets the regulation of Lhx-2 and Lhx-2 is over expressed (3- to 4-fold) in GIC when compared to non-GIC. To determine whether this suppression of Lhx-2 by downregulation of uPA and uPAR is mediated via miR124, we overexpressed miR124 in U87MG and 4910 glioma xenograft GICs and observed that miR124-overexpressed cells failed to establish intracranial tumors in nude mice. Our results demonstrate that the uPA/uPAR system may be involved in GIC maintenance via suppression of miR124.

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 136. doi:1538-7445.AM2012-136
Abstract 1386: Predictive markers of efficacy for an angiopoietin-2 targeting therapeutic in xenograft models
Authors: Triana-Baltzer Gallen B. , Pavlicek Adam , Goulart Ariadne , Huang Hanhua , Woodnutt Gary , Levin Nancy
Published: August 2012

The clinical efficacy of anti-angiogenic therapies has been difficult to predict, and in this era of personalized medicine biomarkers that can predict effect are sorely needed. CVX-060 is an angiopoietin-2 (Ang2) targeting therapeutic, consisting of two peptides that bind Ang2 with high affinity and specificity, covalently fused to a scaffold antibody. The resulting therapeutic has shown promising activity in a phase 1 clinical trial, with several phase 2 clinical programs in progress/planning. In order to optimize the use of this compound in the clinic a set of predictive biomarkers are desired. Here we describe the construction of a predictive model based on the efficacy of CVX-060 in 13 conventional xenograft models, focusing on renal and ovarian cancer. Pretreatment size tumors from each of the models were profiled for the levels of ∼25 protein markers of angiogenesis, SNP haplotype in 5 angiogenesis genes, and somatic mutation status for ∼12 genes implicated in tumor growth and/or vascularization. The markers chosen for evaluation were accumulated from angiogenesis literature and a differential gene expression analysis of CVX-060 treatment-induced changes. CVX-060 efficacy was determined as tumor growth inhibition (TGI%) at termination of each study. A predictive statistical model was constructed based on the correlation of these efficacy data with the marker profiles, and the model was subsequently tested via prospective analysis in 7 additional models The results of this work reveal a range of CVX-060 efficacy in xenograft models of diverse tissue types (0-49% TGI, median = 38%). Notably, we define a subset of 5-7 proteins, the levels of which are predictive of TGI by Ang2 blockade. The majority of these proteins are proximal to the target of CVX-060 and the direction of the associations is such that better efficacy correlates with high levels of target and low levels of compensatory/antagonizing molecules. The SNPs and somatic mutations examined here did not correlate with TGI. Our study describes a systematic approach to the problem of identifying predictive markers of anti-angiogenic affect, and demonstrates the significant ground work necessary to reveal treatment-sensitive predictive markers which can be quantitatively defined. This effort has revealed a set of predictive markers for CVX-060 efficacy that will be further evaluated in ongoing clinical trials.

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 1386. doi:1538-7445.AM2012-1386
Abstract 1395: Docetaxel pretreatment is associated with increased incidence of CNS-metastases in a murine model of HER2-positive breast cancer
Authors: Mischo Axel , Harter Patrick , Mueller Klaus , Kleber Sascha , Zachskorn Cornelia , Renner Christoph , Steeg Patricia S. , Mittelbronn Michel , Pestalozzi Bernhard C.
Published: August 2012

Breast cancer is the most frequent neoplasm in females as it affects 1:10 women. In populations of early breast cancer subsequent brain metastases develop in 5%, while in metastatic breast cancer (mBC), the incidence of CNS-metastases reaches 10-16% [Ann Oncol 2006; 17:935]. The incidence is particularly high (30%-50%) in patients (pts) with HER2-positive mBC [Ann Oncol 2008; 19:1837]. Similarly, the incidence of CNS-metastases is high in pts treated with taxanes for mBC [Ann Oncol 2001; 12:353]. Therefore, we aimed to investigate the role of taxanes on CNS metastases in a murine breast cancer model using the MDA-MB-231BR cell line which has a tropism to metastasize into the CNS upon intracardiac injection [J Natl Ca Inst 2008;100:1092]. Groups of Balb/c nude mice were injected intravenously with docetaxel at a dose of 10mg/kg every 2 weeks for 5 injections (long tax, N=17) or with only 2 similar injections (short tax, N=16) or no injection (control, N=14). Two weeks later MDA-MB-231 BR tumor cells (500′000 cells in 0.1mL PBS) were injected into the left ventricle. Four weeks after that, animals were sacrificed for collection of organs and analysis by histology. The main finding was that metastatic foci were significantly increased in long-tax mice (median number of foci 52, range 0-76) compared to control mice (median number of foci 20, range 0-52; p=0.015). In short-tax animals an intermediate incidence of foci was found (median 30, range 0-79), with a trend, but not a significant difference compared to untreated animals (p=0.7) Histologically we observed a mainly perivascular intraparenchymal infiltration pattern while superficial metastases or a spread to the cerebrospinal fluid was only rarely seen. We therefore assessed changes of the blood-brain barrier (BBB) by ultrastructural approaches. However, we have not found obvious differences in the BBB configuration after taxane treatment. In summary, our results provide evidence that taxane treatment might facilitate spread of breast cancer cells into the CNS, however underlying mechanisms remain to be determined.

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 1395. doi:1538-7445.AM2012-1395
Abstract 1411: Calcium-depending formation of bone metastases in patients with renal cell carcinoma
Authors: Schneider Elke , Roos Frederik , Hampel Christian , Thüroff Joachim W. , Brenner Walburgis
Published: August 2012

Introduction: Although a number of molecules have been implicated in the process of formation of cancer metastases, the organ-selective nature of cancer cells is still poorly understood. Renal cell carcinoma (RCC) often metastasize into bones, a calcium enriched tissue. We analyzed the influence of the calcium sensing receptor (CaSR) in RCC and an enhanced extracellular calcium concentration in processes of metastatic spread. Material and Methods: In 33 matched RCC specimens, 11 non-metastasized, 11 metastasized into the lung and 11 metastasized into bones five years after nephrectomy, CaSR was quantified by RT-PCR and Western blot analyses. To investigate the influence of enhanced extracellular calcium on the formation of metastases, primary RCC cells were isolated from patients developing bone, lung and no metastasis five years after nephrectomy (each 3 cell lines). Cell proliferation after calcium treatment (2.5, 5 and 10 µM) was quantified by BrdU incorporation. Cell migration was determined in a Boyden chemotaxis assay with calcium as chemotactical agent (10 µM). Expression and activity of intracellular signal transduction mediators were analyzed by a human phospho kinase array and Western blot analyses. Results: Compared to non-metastasizing RCC, CaSR mRNA expression was 10.8-fold increased and CaSR protein level was 1.5-fold increased in tumor specimens of patients developing bone metastases. In primary RCC cells with a high bone metastastic potential CaSR expression was 3.7-fold higher compared to non-metastasizing RCC cells. Calcium treatment of bone-metastasizing RCC cells, but not of non-metastasizing cells, induced a concentration dependent enhancement of proliferation. In a Boyden chamber experiment with calcium as chemo attractant, RCC cells from patients with bone metastases showed a 9.4-fold higher migratory potential compared to cells from patients without metastases. In lung metastatic RCC cells migration was only 1.7-fold enhanced. The enhanced proliferation and migration of cells with high potential to build bone metastases seem to be caused by an activation of Akt and a highly increased integrin alpha 5 expression. Additionally, the tumor suppressor PTEN was markedly reduced in tumors metastasizing into bones and after calcium treatment, promoting tumor progression. Conclusion: Our results indicate that formation of bone metastases of RCC is caused by an enhanced expression of the CaSR. Extracellular calcium has a promoting effect on cell migration and proliferation of bone metastasizing RCC cells via Akt signalling and integrin alpha 5.

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 1411. doi:1538-7445.AM2012-1411
Abstract 1412: Progesterone and calcitriol attenuate inflammatory cytokines CXCL1 and -2 in endometrial and ovarian cancer cells
Authors: Kavandi Leyla , Collier Michael A. , Nguyen Huyen , Syed Viqar
Published: August 2012

Chronic inflammation has been shown to play a critical role in growth of cancers. NF-κB is a transcriptional factor that regulates diverse biological processes, including cell proliferation, survival and inflammation. We have previously shown inhibition of endometrial and ovarian cancer cell growth with progesterone and calcitriol. In the present study, we evaluated the effect of progesterone and calcitriol on the expression of inflammatory genes. Using RT-PCR array of inflammatory cytokine and receptor genes, we found two inflammatory cytokines, CXCL1 and -2 (GRO-A and -B), downregulated by both treatments. PCR array results were validated by Western blotting. Knockdown of NF-kB resulted in a reduced expression of CXCL1 and -2 and the inhibitory effect of progesterone and calcitriol on the expression of cytokines was abrogated in NF-kB-silenced cancer cells. Silencing of IkBα increased the expression of CXCL1 and -2 in cancer cells, which can be attributed to the increased activation of NF-kB-p65, caused by the lack of its inhibitor. Progesterone and calcitriol-induced inhibition was abolished in IkBα-knockdown cells. Our results demonstrate that suppression of IkBα phosphorylation by progesterone and calcitriol contributes to the reduced expression of CXCL1 and -2. We investigated the effect of CXCL1 and -2 downregulation on metastasis-promoting genes and found a marked inhibition of CXCR4, COX2 and matrix metalloproteinases (MMP2 and MPP-9). Overall, our results indicate that progesterone and calcitriol downregulate NF-kB in cancer cells, leading to the suppression of metastasis, thus providing the molecular basis for the treatment of endometrial and ovarian cancer patients with progesterone and calcitriol.

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 1412. doi:1538-7445.AM2012-1412
Abstract 1414: Regulation of tumor cell trafficking by chromogranin A
Authors: Dondossola Eleonora , Crippa Luca , Colombo Barbara , Ferrero Elisabetta , Corti Angelo
Published: August 2012

Chromogranin A (CgA) is an acidic glycoprotein present within secretory vesicles of many normal and neoplastic endocrine and neuroendocrine cells. CgA is abnormally released in circulation in patients with neuroendocrine and non neuroendocrine tumors, as in a subpopulation of patients with non small cell lung cancer (NSCLC), with important prognostic implications. We have previously demonstrated that CgA can preserve the barrier function of the vascular endothelium. This prompted us to investigate the role of circulating CgA on metastasis development and tumor self-seeding, the ability of tumor cells to re-infiltrate their tumor of origin. Using mouse mammary adenocarcinoma models we observed that CgA inhibits both tumor self-seeding and organ colonization. Studies of the mechanism of action showed that CgA can inhibit the shedding of cancer cells in circulation from primary tumors, the re-infiltration of tumors and the colonization of lungs by circulating tumor cells. Pharmacological modulation of CgA further affected the development of lung colonies by tumor cells. We obtained evidences to suggest that CgA can inhibit the transendothelial migration of cancer cells, by preserving the VE-cadherin-dependent endothelial barrier function. These findings point to a role of circulating CgA as a regulator of tumor cell trafficking from tumor-to-blood and from blood-to-tumor/normal tissues. Inhibition of the multidirectional trafficking of cancer cells in normal and neoplastic tissues may represent a novel strategy to reduce cancer progression.

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 1414. doi:1538-7445.AM2012-1414