bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 769: Effect of cucurbitacin B, a medicinal herb extract from Trichosanthes cucumerina, on Wnt signaling pathway in breast cancer cells
Authors: Dakeng Sumana , Duangmano Suwit , Jiratchariyakul Weena , Bögler Oliver , Patmasiriwat Pimpicha 
Published: February 2011

Abstract
The cucurbitacins are tetracyclic triterpenes found in plants of the family Cucurbitaceae. Cucurbitacins have been shown to have anticancer and anti-inflamatory activities. In this study, we investigated the anticancer activity of cucurbitacin B and dihydrocucurbitacin B extracted from Thai medicinal plant Trichosanthes cucumerina. A mixture compound of cucurbitacin extracts containing cucurbititacin B and dihydrocucurbitacin B in an 8:1 molar ratio was dissolved in 10% dimethylsulfoxide and diluted with DMEM to the desired concentrations prior to use. Cell viability was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Results indicated that mixture of cucurbitacin B (mixture CuB) from T. cucumerina have cytotoxic effect on breast cancer cell lines, SKBR-3, MCF-7, HBL-100 and T47D with the IC50 of 4.60 µg/ml, 88.75 µg/ml, 55.46 µg/ml and &gt;100 µg/ml respectively. Flow cytometry for cell cycle analysis was performed after treated cells with mixture CuB. We found that growth inhibition effect was attributed at G2/M phase arrest and apoptosis. Western blotting analysis showed increase PARP cleavage and decrease Wnt associated signaling molecules including β-catenin, galectin-3, cyclin D1 and c-Myc. We also found that mixture CuB inhibit nuclear-translocation of β-catenin and galectin-3 when compared with vehicle control cells. The depletion of β-catenin and galectin-3 in the nuclease was confirmed by cellular protein fractionation method. TCF promoter, a downstream signaling of Wnt was also disrupted in cucurbitacin B treated cells. TCF promoter assay system was used for assessment of TCF promoter activity. The relative luciferase activity was reduced when we treated cells with mixture CuB for 24 hours. Our data indicate that cucurbitacin B and their derivative dihydrocucurbitacin B from T. cucumerina induce apoptotic cell death and exhibit growth inhibitory effect via inhibition of Wnt signaling pathway.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 769.
Abstract 78: Role of 12-lipoxygenase in regulation of ovarian cancer cell proliferation and survival
Authors: guo austin M. , Liu Xiuli , Al-Walhab Zaid , Maddippati Rao , Scicli A. Guillermo , Munkarah Adnan 
Published: February 2011

Abstract
Enzymes involved in eicosanoid synthesis have previously been implicated in the progression of various types of cancer, including ovarian carcinomas. Most of the current data in ovarian cancer have focused on cyclooxygenase-2 (COX-2). It is not known whether the main human 12-hydroxyeicosatetrenoate (12-HETE) generating enzyme, 12-lipoxygenase (12-LOX), contributes to the regulation of growth of ovarian cancer cells. In the present study, we examined the function of the 12-lipoxygenase pathway in the regulation of proliferation and survival of three epithelial ovarian cancer (EOC) cell lines. We found that all three EOC cells express high level of the mRNA encoding for 12-LOX, using RT-PCR. These cells also contain significant amount of immuno-reactive 12-LOX protein as revealed by western blot. Liquid chromatography/Mass Spectrometry (LC/MS) analysis confirmed that all three EOC produce 12-HETE when challenged with arachidonic acid (AA). N-benzyl-n-hydroxy-5-phenyl-pentanamide (BMD-122) is a specific 12-LOX inhibitor. Exposure of the three EOC cell lines to an increasing dose of the 12-LOX inhibitor, BMD-122 (1 μM, 10 μM and 50 μM), resulted in a dose-dependent inhibition in cell proliferation (75±9%, 51±6% and 26±4% of untreated control, respectively), measured by MTT assay in vitro. Furthermore, treatment with 10 μM BMD-122 was associated with up to ∼30% increases in the apoptosis of these EOC cells within 24 hr, measured by Annexin-V flow cytometry. This apoptosis seems to be mediated through a mechanism independent of PI-3 kinase since the expression of phosphorylated Akt was not affected by BMD-122 treatment. Furthermore, we found that BMD-122 treatment had no effect on the three EOC cell lines ability to invasion using Matrigel assay. Taken together, these data suggest that 12-LOX is involved in regulation of ovarian cancer progression and survival, an important and novel finding. Inhibition of 12-LOX may have potential as an adjuvant therapy in ovarian cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 78.
Abstract 781: Bioconversion of curcumin and its analog by Beauveria bassiana ATCC 7159
Authors: Martin Glenroy , Ebert Tamara , McPhee Jennifer , Mercer Bettie , Williams Jesica 
Published: February 2011

Abstract
Curcumin (1), a chemopreventive agent from turmeric (Curcuma longa L.) was incubated with the microbial cultures of Beauveria bassiana ATCC 7159. Bioconversion of [(1E, 6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione] (1) with B. bassiana afforded five major metabolites: 1,7-bis(4-hydroxyphenyl)heptane-3,5-dione (2), 3-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)hept-1-ene (3), 1,2,6,7-diepoxy-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-3,5-diol (4), 5-hydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl)hept-1-ene (5) and 1,7-bis(4-hydroxy-3-methoxyphenyl)heptan-3,5-diol (6). The biotransformation of the synthetic analog 1,7-bis(4-hydroxy-3-methoxyphenyl)heptane-3,5-dione (7) by B. bassiana afforded the reduced diol 1,7-bis(4-hydroxy-3-methoxyphenyl)heptan-3,5-diol (6) as the only metabolite.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 781.
Abstract 782: Characterization of genetic changes in oligodendroglial tumors including glioblastomas with oligodendroglial component
Authors: Klink Barbara , Krex Dietmar , Gerlach Eva-Maria , Schlingelhof Ben , Hackmann Karl , Patt Stephan , Schrock Evelin 
Published: February 2011

Abstract
In contrast to astrocytic tumors including glioblastomas (GBM), oligodendrogliomas (O) show a better prognosis and increased responsiveness to chemotherapy. Interestingly, a small subgroup of glioblastomas contains areas with histological features of oligodendroglial differentiation. In the current WHO classification (2007) these tumors are now included as a variant of glioblastomas and are called glioblastomas with oligodendroglial component (GBMO). However, definitive diagnostic criteria still do not exist.
In this study we used a genome wide approach (chromosomal comparative genomic hybridization (CGH) and molecular karyotyping) in combination with interphase fluoreszence in situ hybridization (FISH) (regions 1p, 1q, 7q, 10q, 17p, 19q, cen18, 21q) to genetically characterize GBMO and “classical” O. We analyzed the oligodendroglial and the astrocytic glioblastoma parts of 13 GBMO separately by chromosomal CGH of microdissected paraffin embedded tumor tissue and interphase FISH on paraffin sections. Furthermore, freshly frozen material of six oligodendrogliomas (4 WHO grade II, 6 WHO-grade III) was examined genome wide by using the standard Agilent 244A chip.
We identified four distinct genetic subtypes in GBMO: an “astrocytic” subtype (9/13) characterized by +7/-10; an “oligodendroglial” subtype with −1p/-19q (1/13); an “intermediate” subtype showing +7/-1p (1/13), and an “other” subtype having none of the former aberrations typical for gliomas (2/13). In contrast, most “classical” O (5/6) showed combined 1p/19q deletion, corresponding genetically to the “oligodendroglial” subtype. Four of the O with 1p/19q codeletion showed additional aberrations (e. g. −4, −14, −9, −18). The different histological tumor parts of GBMO revealed common genetic changes in all tumors and showed additional aberrations specific for their oligodendroglial and astrocytic parts. These findings demonstrate the monoclonal origin of the tumor followed by the formation of the astrocytic and oligodendroglial components. We are currently evaluating the array CGH results of six oligodendrogliomas, which will also be presented.
The diagnostic determination of these genetic signatures may allow for a better prognostication of the patients. Our results underline the importance of molecular cytogenetic analyses to supplement the histological diagnosis of gliomas.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 782.
Abstract 784: Plasmacytoid urothelial carcinoma: Molecular and histopathological characterization of a rare variant of bladder cancer
Authors: Stoehr Robert , Keck Bastian , Rogler Anja , Hofstaedter Ferdinand , Fritsche Hans-Martin , Lehmann Jan , Montironi Rodolfo , Sibony Mathilde , Wullich Bernd , Hartmann Arndt 
Published: February 2011

Abstract
AIM: The plasmaycytoid urothelial carcinoma (PUC) is a rare variant of urothelial carcinoma. To date only few cases have been reported and molecular data are not published yet. Therefore, we analysed clinical, histopathological and molecular features in a series of 32 PUC.
METHODS: Expression of CK7, CK20, PAN-CK, p53, Ki67, CD138, E-cadherin and beta-catenin was analysed immunohistochemically on a tissue microarray. UroVysion FISH was performed to asses ploidy status. After microdissection and DNA isolation, FGFR3 and PIK3CA mutation status were determined using SNaPshot analysis. P53 mutation analysis was performed by direct sequencing of exons 5-9. Clinical data available from 16 PUC were compared to 269 patients with conventional locally advanced urothelial bladder cancer within a randomized multicenter trial (AUO-AB 05/95).
RESULTS: PUC were usually diagnosed in advanced pathologic stage (64% pT3, 23% pT4) with 60% metastatic disease. Overall survival of PUC treated with cystectomy and adjuvant chemotherapy was lower than of patients with conventional locally advanced bladder cancer (23.3 vs. 49.9 months, p=0.051). 87% of PUCs showed loss or strongly reduced expression of E-cadherin. Beta-catenin was completely absent in 22.5% and associated with loss of E-cadherin (p=0.033). 16.7% of PUC showed a nuclear staining for beta-catenin, which was more frequent in E-cadherin negative PUC. CK20 expression was abnormal in all cases. 22.6% of PUCs were negative for CK7. 97% of PUC displayed a positive PAN-CK staining. 78% coexpressed CD138. Nuclear accumulation of P53 was found in 68.4% of E-cadherin negative PUC, but only in 27.2% of E-cadherin positive ones. p53 mutations were found in 9/31 cases. 69% of PUC showed increased Ki67 labelling index (&gt;15%). FISH displayed PUC as highly aneuploid tumors. Relative deletion of 9p21 was found in 70%. No mutations were found in FGFR3 and PIK3CA.
CCONCLUSIONS: PUC showed molecular and histopathologic features of aggressive bladder cancer. Prognosis in locally advanced PUC treated with adjuvant chemotherapy seems to be unfavourable in comparison to conventional bladder cancer. Loss of E-Cadherin is present in the majority of PUC and is associated with a higher degree of p53 and beta-catenin accumulation, which is in line with the high metastatic potential of PUC. CD 138 is not suitable for differential diagnosis to plasmocytoma. PAN CK is an important diagnostic tool for clarification of the epithelial origin of PUC as there are cases negative for CK7 and CK20.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 784.
Abstract 789: Determination of MAPK and Wnt pathway activity in esophageal cancer
Authors: Shannon Nicholas B. , Peters Christopher J. , Rees Jonathan R. , Hardwick James S. , Zhang Chunsheng , Hardwick Richard H. , Fitzgerald Rebecca C. 
Published: February 2011

Abstract
Introduction:
Gastro-esophageal adenocarcinomas are rapidly increasing in incidence and have a five year survival of less than 14%. Analysis of signaling pathways in gastro-esophageal adenocarcinomas can be used as a route to identifying possible therapeutic targets.
Aims and methods:
We had already identified MAPK and Wnt pathways as important in this disease. The aim of this work was to identify biomarkers of pathway activation for future application to primary tumors for selection of targeted therapies.
Three cell lines (CP-B, CP-C, CP-D) without constitutive activation were stimulated with exogenous ligands for the Wnt or MAPK or pathways. The TGFβ pathway was also stimulated as a negative control to test biomarker specificity. The expression of potential biomarkers of transcriptional activity was then profiled using real time PCR. To determine applicability of these markers to human tissue the expression of these markers was assessed in an expression array data set consisting of 75 gastro-esophageal adenocarcinomas.
Results:
Out of 8 (MAPK) and 13 (Wnt) potential markers, three markers were selected as specific for the MAPK pathway (EGR-1, JUN and FOSL1) or the Wnt pathway (BMP4, TCF7, LEF1). All markers had upregulation of expression of at least 2 fold that were specific to activation of the relevant pathway (Table 1).Table 1.Fold changes of expression upon activation by stimulation with exogenous ligandsMarkerEGF (MAPK pathway)LiCl (Wnt pathway)TGFβ (TGFβ pathway)P-value (ANOVA)EGR-1130.50.91.3&lt; 10-4JUN3.31.21.0&lt; 10-4FOSL111.02.60.6&lt; 10-4BMP42.18.31.5&lt; 10-4LEF10.523.30.5&lt; 10-4TCF72.44.80.7&lt; 10-4
Next we assessed the expression of selected markers in 75 gastro-esophageal adenocarcinomas for which expression profiling data was available. In these tumors 27 (36%) had activation of the MAPK pathway alone, 10 (13%) had activation of the Wnt pathway alone, and a further 12 (16%) had activation of both pathways.
Conclusions:
We have determined a set of markers which can be used to assess pathway activation of the MAPK and Wnt pathways in gastro-esophageal adenocarcinomas. External validation of these pathway activation markers is ongoing using multiplex PCR as a prelude to a clinical trial of targeted therapy.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 789.
Abstract 79: The compartmentalization of folate metabolism depends on two isoforms of folylpoly-γ-glutamate synthetase and the mitochondrial folate transporter
Authors: Lawrence Scott A. , Moran Richard G. 
Published: February 2011

Abstract
The reactions of folate metabolism are distributed between the cytosolic and mitochondrial compartments of mammalian cells and both components are essential for mammalian cell survival. The enzyme folylpoly-γ-glutamate synthetase (FPGS) has been shown to be essential for the intracellular retention of folates as polyglutamate derivatives. The fpgs gene produces two transcripts, one for an isoform with a leader peptide targeting it to the mitochondrial matrix, and the other is transcribed without this sequence, so that the encoded isoform remains in the cytosol. Transfer of folates into the mitochondrial matrix is mediated by an inner mitochondrial membrane transport protein, the mitochondrial folate transporter (MFT). Either deletion of the MFT or of mitochondrial FPGS is cytotoxic and renders the cell auxotrophic for glycine; the mechanism of this effect is far from apparent. To better understand this compartmentalization, we generated cell lines that express only the cytosolic or mitochondrial isoform of FPGS under tetracycline-inducible control in FPGS-null AuxB1 cells. Western blot analysis of the FPGS protein in these cell lines indicate that both FPGS isoforms are expressed near endogenous levels in the presence of doxycycline and that each cell line solely expresses the FPGS isoform in the respective compartment. Clonogenic survival of each doxycycline-induced cell line demonstrates a cellular requirement for each FPGS isoform. Additional studies showed that cells complemented with cytosolic FPGS only accumulated and retained folates in the cytosol but not mitochondria, leading us to conclude that folate polyglutamates are not substrates for the MFT. In addition, expression of only the mitochondrial form of FPGS resulted in cells that progressively trapped folates in the mitochondria, leading us to the concept that folate polyglutamates do not escape the mitochondria. However, cytosolic folates were observed in cells expressing only mitochondrial FPGS. We are currently using these cell lines to study the transfer of folates across the mitochondrial membrane and polyglutamation in each compartment.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 79.
Abstract 796: Molecular characterization of circulating tumor cells using fluidigm biomark dynamic array
Authors: Jiaqi Huang , Brohawn Philip , Bao Haifeng , Burke Patricia , Chen Xiaoru , Zhu Wei , LaVallee Theresa , Jallal Bahija , Yao Yihong 
Published: February 2011

Abstract
Circulating tumor cells (CTCs) are cancer cells derived from a primary tumor and/or its metastases that circulate in the peripheral blood. The number of CTCs in whole blood of cancer patients has been shown to have clinical relevance with respect to patient prognosis. There is, in addition to CTC enumeration, great interest in the molecular characterization of isolated CTCs. However, there are challenges for molecular characterization of CTCs with commonly used methodologies. The number of CTCs is extremely low and despite the depletion of leukocytes during cell enrichment, the CTC-enriched fractions still include numerous leukocytes which strongly impact CTC-specific gene expression profiling. CellSearch™ CTC test is a FDA approved diagnostic test for enumeration of CTCs. The fixation reagents in the CellSave tube allow processing of samples for several days post-collection, making analysis of clinical samples more practical compared to near-same day processing required for samples collected in EDTA tubes. However, the fixation decreases the mRNA signal and assay sensitivity. We have established a protocol which overcomes these hurdles and allows the profiling of select transcripts of interest in CTCs. By applying a proteinase K treatment and Trizol LS lysis prior to RNA isolation, we were able to overcome the impact of fixation on RNA isolation. RNA was isolated from the treated lysate using a commercially available kit applicable to low cell numbers. Use of this kit allowed us to utilize all of the isolated RNA in the cDNA step. Superscript III cDNA synthesis was then carried out according to the manufacturer's protocol. Target genes of interest were then pre-amplified using the Life Technologies established methodology and reagents. The resulting volume of pre-amplified cDNA was concentrated to a set volume to maximize the amount of transcript profiled. Genes of interest were profiled on the Fluidigm Biomark 48.48 Array. mRNA expression analysis was performed on genes which are not expressed in a pure leukocyte population to ensure that any signal detected was from the CTCs, and not from the background leukocyte population. Using this methodology, we succeeded in performing quantitative gene expression analysis on genes such as PDGFRα on as few as five tumors cells (MG63) fixed in CellSave tubes and spiked into a background environment of contaminating leukocytes (typically 1000 or more) pulled down from normal blood using the CellSearch system. These methods could potentially be used not only for greater understanding of the biology and metastatic potentialof CTCs
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 796.
Abstract 800: Novel GMDS mutation and clinical estimation of fucosylation in several kinds of human cancer
Authors: Moriwaki Kenta , Tsuda Saori , Shinzaki Shinichiro , Furukawa Yoichi , Miyoshi Eiji 
Published: February 2011

Abstract
Oligosaccharides are some of the most important factors in the posttranslational modification of proteins and lipids. It is well known that oligosaccharide structures change during malignant transformation. Fucose is a type of constituent of oligosaccharides, and is notably associated with cancer and inflammation. Fucosylation is catalyzed by several kinds of fucosyltransferases, the GDP-fucose synthetic enzymes, and GDP-fucose transporter. Recently, we found a mutation of the GMDS gene, which involved in the synthesis of GDP-fucose, in a colon cancer cell line, HCT116, leading to a virtually complete deficiency of fucosylation. Down-regulation of cellular fucosylation rendered the cells resistant to tumor necrosis factor-related apotosis-inducing ligand (TRAIL) -induced apoptosis, resulting in the escape from natural killer (NK) cell-mediated tumor immune surveillance. Thus, GMDS and fucosylation are thought to be regulatory factors for TRAIL-induced apoptosis and tumor immune surveillance.
In the present study, we searched mutation of the GMDS gene in human cancer cell lines and various kinds of cancer tissues. The aberrant transcripts with deletions of exons 2-4 and 5-7 were detected in the cDNA of LS174T and NCI-H716 cells, respectively. Since these mutations were heterozygous, the deficiency of fucosylation was not observed. In a gastric choriocarcinoma cell line, SCH, only short transcript with deletion of exons 2-4 was detected. This cell line showed a marked decrease in the fucosylation level. PCR and subsequent direct sequencing analyses using 100 human colon cancer tissues identified mutations in two cases. One of the two was c.739 C&gt;T (p.R247X) in exon7, and the other was c.345+5 G&gt;A (IVS4+5) in intron 4. Furthermore, RT-PCR analysis identified aberrant transcripts in 5 of 10 ovarian cancer tissues. All mutant GMDS genes found in this study had no enzymatic activity.
Subsequently, the level of fucosylation in many colon cancer tissues was investigated by immunohistochemical analysis using two types of fucosylated oligosaccharide-specific lectin. The decrease of fucosylation was observed in certain cases. Especially, core-fucosylation was decreased in colon cancer with lymph node metastasis.
These results suggest that cancer progression in certain patients with colon cancer is dependent on de-fucosylation. Thus, fucosylation is an attractive target for cancer diagnosis and therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 800.