bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2672: Characterization of Hsp90/Cdc37 interaction and their critical residues using luciferase fragment complementation imaging
Authors: Jiang Yiqun , Bernard Denzil , Yu Yanke , Zhang Tao , LI Yanyan , Fu Xueqi , Wang Shaomeng , Sun Duxin 
Published: February 2011

Abstract
The purpose of this study is to investigate Hsp90/Cdc37 interaction and identify residues critical for the interaction using real-time luciferase complementation imaging and computational modeling. Molecular imaging using a split Renilla luciferase fragment complementation (SRL-FC) was established to monitor real-time Hsp90/Cdc37interactions in living cells. Computational modeling and molecular dynamics simulations were used to evaluate the interaction interface of Hsp90/Cdc37 complex and critical residues in the interaction interface were identified for mutagenesis. Western Blotting and co-immunoprecipitation were used to determine protein expression and complex formation. The molecular imaging in living cells showed that Hsp90 and Cdc37 helped the complementation of N-terminus of Renilla luciferase (fused with Hsp90; N-RL-Hsp90) and C-terminus of Renilla luciferase (fused with Cdc37, Cdc37-C-RL) with 170-fold higher luciferase activity than controls without Hsp90/Cdc37. This suggests that the SRL-FC is sensitive and specific to monitor the interaction of full length human Hsp90 and Cdc37. Molecular dynamics simulations revealed that the interface was constituted by a series of residues interacting through hydrophobic or polar interactions. Mutagenesis confirmed that mutations in Hsp90 (A121N, Q133A, F134A) and mutations in Cdc37 (M164K, R167A) led to the loss of complementation of N-RL-Hsp90 and Cdc37-C-RL (5 to 10-fold lower luciferase activity compared to wild-type), indicating these residues are critical in Hsp90/Cdc37 interactions. In comparison, mutations in Hsp90 (E47A, S113A) and mutations in Cdc37 (A204E) only decreased 50% of the complementation of N-RL-Hsp90 and Cdc37-C-RL, which suggests that these residues contribute less to Hsp90/Cdc37 interaction. In contrast, mutations of Hsp90 (R46A, S50A) and mutation in Cdc37 (L165H) did not change the complementation of N-RL-Hsp90 and Cdc37-C-RL, indicating that these residues did not contribute to Hsp90/Cdc37 interaction. The data suggests that molecular imaging using split Renilla luciferase fragment complementation (SRL-FC) can be used to identify the critical residues in Hsp90/Cdc37 interaction. Although Hsp90/Cdc37 interactions are through both hydrophobic and polar interactions, mutation in a single amino acid residue, including Ala121, Gln133, Phe134 in human Hsp90 and Met164, Arg167 in human Cdc37, is sufficient to disrupt Hsp90/Cdc37 interaction.
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 2672.
Abstract 2675: Characterization of human thioesterase 2 and its potential for breast cancer drug discovery
Authors: Ritchie Melissa K. , Johnson Lynnette C. , Watts Laura A. , Kridel Steven J. , Lowther W. Todd 
Published: February 2011

Abstract
Human thioesterase II (TE2) may limit the anti-tumour potential of fatty acid synthase (FASN) inhibitors designed to target endogenous thioesterase (TE1) domain of FASN in breast cancer. Recent studies have firmly established the anti-tumour effects of TE1-targeted FASN inhibition via the drug Orlistat in prostate tumour xenografts, and current studies are revealing similar cytotoxic effects in breast cancer cell lines. Both TE1 and TE2 act on the acyl carrier protein (ACP) domain of FASN to liberate fully saturated, long-chain or medium-chain fatty acids, respectively. Therefore, TE2 may rescue fatty acid synthesis on TE1 inhibited FASN in cancer cells, thus highlighting TE2 as a potential new target for cancer therapy. Encouraged by the developments in TE1 inhibitors and by the expression of TE2 in breast cancers, we are now investigating the biochemical and structural properties of recombinant TE2 in order to define the factors that determine substrate specificity. In addition, the expression profile of endogenous TE2 in several breast cancer cell lines is under investigation. The long-term goal of this project is to determine whether TE2 is a viable target for the development of breast cancer specific anti-tumour therapies and/or whether a combined drug discovery strategy against both thioesterases must be pursued. Here we report the kinetic profiling of human TE1 and TE2 using model substrates and select β-lactone inhibitors. As well, we report immunoblot analyses of endogenous TE2 in MCF 7, SKBr 3, and MDA 231 breast cancer cell lines. Finally, we report the current progress of determining the crystal structure of recombinant TE2. Our results show lower β-lactone inhibitor potency against TE2 compared with TE1, as well as differential expression of endogenous TE2 in breast cancer cell lines. These results suggest that TE2 may need to be specifically targeted in breast cancer in order to fully realize the anti-tumour potential of FASN inhibition.
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 2675.
Abstract 2679: Virtual screening of small molecule inhibitors towards the tumor suppressor p53 to overcome radioresistance of nasopharyngeal carcinoma cells
Authors: Eichhorn Tolga , Efferth Thomas 
Published: February 2011

Abstract
The tumor suppressor p53 belongs to the most frequently mutated genes in cancer, including nasopharyngeal carcinoma (NPC). p53 is over-expressed in NPC, but the mutation rate is lower than in other tumor types. Therefore, it can be hypothesized that p53 might be involved in repair of DNA-damage after radiotherapy and causes recrudescence. Hence, the aim of the present investigation was to identify small molecule inhibitors for p53 to inhibit DNA repair after ionizing radiation of NPC.
Approximately 260.000 2D-structures from NCI database were downloaded and different tautomers and charged stages between pH 5 and 9 were calculated for each compound. A total number of 906.587 drugs in different 3D-conformations have been generated for our virtual screening approach. By means of the Glide software and three-dimensional p53 structure based on the published crystal structure 1TUP from PDB database, 25 compounds have been identified as candidates to bind to the DNA-binding domain of p53. The best evaluated ligand (consisting 26 atoms) had the assigned binding energy of −8.41 kcal mol-1 in Glide. This outstanding binding was confirmed by using AutoDock which has result in −10.57 kcal mol-1. In comparison, the binding of PRIMA-1 as experimentally validated drug showed a binding energy of −6.30 kcal mol-1 after blind docking procedure with subsequent refinement.
As a next step, the compounds identified in silico have been experimentally verified in vitro by means of Plasmon resonance spectroscopy. To prove, whether inhibition of DNA-binding of p53 by these small molecule inhibitors affect the response of the NPC cell line CNE1 towards ionizing radiation, the cytotoxicity of 0.5 – 16 Gy has been measured with and without addition of these small molecule inhibitors by XTT assays. Indeed, the addition of some of the identified candidate compounds were able to increase the cytotoxic effects of ionizing radiation.
The present investigation indicates that the combination of ionizing radiation with novel small molecule inhibitors for p53 increased the killing rate of CNE1 cells. These data may have important implications for radiochemotherapy of NPC in the future.
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 2679.
Abstract 2688: The expression of lung resistance protein (LRP) in patient ascites predicts platinum resistance in advanced epithelial ovarian carcinoma
Authors: McNally Lacey R. , Frederick Peter J. , Bevis Kerri S. , Sellers Jeffrey C. , Della Manna Debbie L. , Straughn J Michael , Stockard Cecil R. , Grizzle William E. , Buchsbaum Donald J. 
Published: February 2011

Abstract
The human lung resistance protein (LRP) is a multidrug resistance protein that has been shown to be overexpressed in numerous chemotherapy resistance models. The precise role of LRP in platinum-resistant epithelial ovarian cancer (EOC) has yet to be fully elucidated. In this study, we sought to determine LRP expression in the tumor cells of ovarian cancer patient ascites of chemotherapy-naïve patients with advanced EOC.
After IRB approval was obtained, patient ascites was collected and evaluated for platinum sensitivity, and LRP expression using western blot and immunohistochemistry (IHC). Both patient ascites and primary tumor were treated with 50 uM carboplatin for 48h and cell viability was analyzed. LRP, activated Caspase 3, and tunnel were analyzed using IHC on samples treated with 50uM carboplatin and untreated.
A total of 27 patients with advanced-stage EOC undergoing primary cytoreductive surgery at the University of Alabama at Birmingham were consented for the study. Ten patient ascites samples were deemed adequate for analysis. Of these, 3/10 (30%) had increased LRP levels in ascites samples demonstrated by strong IHC staining with capping and confirmed by western blot. These patient samples were platinum resistant as verified by cytotoxicity analysis. Two ascites specimens (20%) lacked LRP expression by IHC and western blot and demonstrated platinum sensitivity on cytotoxicity. Five ascites specimens (50%) displayed intermediate LRP staining and platinum cytotoxicity. Tumor cells in ascites treated with carboplatin had an inverse correlation of LRP staining and activated Caspase 3.
There is a strong correlation between LRP expression in the ascites of patients with advanced EOC and platinum resistance in this ex vivo model. This protein may therefore predict poor prognosis disease unlikely to respond to adjuvant platinum therapy. Specifically targeting the LRP in a clinical model may reverse carboplatin resistance.
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 2688.
Abstract 2694: DNA copy number aberrant profiles predict complete pathologic tumor response to preoperative chemoradiation therapy in locally advanced rectal cancer
Authors: Chen Zhenbin , liu Zheng , Wu Xiwei , Li Wenyan , Garcia-Aguilar Julio 
Published: February 2011

Abstract
Purpose: The treatment of locally advanced rectal cancer includes chemoradiation (CRT) followed by surgical resection. The response of rectal cancers to the CRT is variable. The finding that up to 25% of tumor seem to be totally eradicated by the CRT has raised the question as to whether surgery can be avoided in these patients. Findings predictors or response or resistance to CRT could potentially change the treatment paradigm of rectal cancer patients. In this study we explored DNA copy number alterations as predictors of rectal cancer response to preoperative CRT.
Patients and Methods: High-density oligonucleotide-based comparative genomic hybridization (human 244k aCGH kit, Agilent Cor.) was employed to identify the DNA copy number alterations in 89 pretreatment tumor biopsy specimens and paired normal surgical specimens collected from stage II and III rectal cancer patients treated in a prospective multi-institutional study. Data analysis was performed using Nexus Copy Number and Ingenuity Pathway Analysis. A pathologic complete response (pCR), defined as absence of cancer cells in the surgical specimen, was used as clinical end-point.
Results: A total of 25 (28%) patients had a pCR. The pCR rate was higher among tumors with low fractional genomic alterations (FGA, ≤21%) (12/26 or 46.1%) compared to tumors with high FGA (&gt;21%) (13/63 or 20.6%) (p&lt;0.02). Gain or losses of individual chromosome segments containing individual candidate genes and miRNAs in tumors with or without pCR are presented in table 1.Chrom. LocationCandidate GeneChangepCRNon- pCRp-valuechr18q21.2SMAD4loss52%82.8%0.0059chr.8p12EIF4EBP1loss8%35.9%0.0085chr.17q12ERBB2gain4%29.7%0.0098chr.16p12.1PRKCBgain4%29.7%0.0098chr.7p15.2hsa-mir-148again32%58.7%0.0352
Conclusion: This study shows that DNA copy number aberrations, measured as a FGA and individual chromosome regions gain and loses, may be predictors of rectal cancer resistance to CRT. This information could be used to select patients that may not benefit from a non-surgical treatment.
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 2694.
Abstract 270: Modulation of EGFR localization and transcriptional activity
Authors: Bitler Benjamin G. , Goverdhan Aarthi , Schroeder Joyce 
Published: February 2011

Abstract
The triple negative (ER-, PR-, Her2-) or basal-like breast cancers are the most difficult forms of breast cancer to treat, because the cancer is often chemoresistant and highly metastatic. As these cancers progress from early to late stage the 5-year survival rate drops dramatically. Many proteins are responsible for this progression, including MUC1 and the Epidermal Growth Factor Receptor (EGFR). MUC1 is a heavily O-glycosylated transmembrane protein that is over expressed in 90% of all breast and ovarian cancers. In normal cells, MUC1 is only found on the apical surface of epithelium, but in cancer, MUC1 is found to be mislocalized. As a result, MUC1 plays a role in promoting tumorigenesis and metastases, by interacting with other oncogenes such as EGFR. EGFR is overexpressed in about 60% of the most aggressive form of breast cancer (triple negative). In an EGFR-dependent mouse model of breast cancer, the loss of Muc1 directly correlates to the loss of cyclin D1. Additionally, blocking MUC1 and EGFR interactions significantly inhibited tumor progression in a xenograft and s spontaneous mouse model of breast cancer. Recently, a novel trafficking pathway of EGFR has been discovered, which was described as EGFR trafficking to the nuclear compartment. Following entry into the nucleus EGFR can act in an oncogenic manner by aberrantly activating transcription of genes such as, cyclin D1 and v-myb. In this study we discovered a novel regulator function of MUC1 in EGFR trafficking and transcriptional activity. We found that MUC1 expression in basal-like breast cancer promotes the accumulation of nuclear EGFR and mediates EGFR's interaction with chromatin. Additionally, we determined that EGFR's interaction with the cyclin D1 promoter and upregulation of the cyclin D1 protein was MUC1-dependent. The main focus of anti-EGFR therapeutics have been its tyrosine kinase activity, however this study reveals that EGFR's subcellular localization should also be considered in the development of future cancer therapeutics.
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 270.
Abstract 2720: Glycosylation of Mucins Expressed in Pancreatic Adenocarcinoma
Authors: Remmers Neeley , Rothermund-Franklin Christy , Caffrey Thomas , Hollingsworth Michael A. 
Published: February 2011

Abstract
Glycosylation of mucins expressed by pancreatic adenocarcinoma
There were an estimated 42,470 new cases and 35,240 deaths due to pancreatic adenocarcinoma (PA) for 2009, ranking PA as the third leading cause of cancer related death. Virtually all long term survivors are diagnosed early but only 7% are detected in the early stages of disease thus highlighting the need for an early diagnostic test. CA19-9, the only current clinical serum-based assay used to monitor PA, detects an oligosaccharide blood group antigen Sialyl Lewisa (sLeA) with the structure (Neu5Acα2-3Galβ1-3[Fucα1-4]GlcNAc), which can be present on glycolipids or as an O-linked oligosaccharide on glycoproteins, and is expressed at relatively low levels on the surfaces of normal epithelial cells. The full spectrum of proteins on which the CA19-9 epitope is expressed has never been defined. CA19-9 is differentially expressed on mucins, which are extensively glycosylated. The CA19-9 test lacks high specificity for PA, as CA19-9 is commonly elevated in patients with benign chronic pancreatitis and other malignant diseases. Different epithelial tissues and cell types produce distinct combinations of mucin-type core proteins, and the expression patterns of these are generally altered during different disease processes including malignant transformation. Mucin expression is deregulated in cancer, which results in aberrant expression and glycosylation of mucin core proteins. We employed immunohistochemistry and immunofluorescence to show the unique mucin and glycosylation patterns seen in tissue specimens from patients diagnosed with PA. Deglycosylation of samples immunoprecipitated from cancer cell lines and pancreatic cancer patient tissue specimens was used to enable detection of mucin core proteins. Immunoaffinity chromatography was used to isolate CA19-9 from patient ascites and serum samples and mass spectrometry was used to analyze the core proteins. Results show that pancreatic cancers expressing the mucin core proteins MUC1, MUC4 and MUC5AC carry the CA19-9 epitope. Preliminary results suggest that a strategy to detect the CA19-9 tumor antigen and its cognate mucin core protein(s) will improve the performance characteristics of this assay.
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 2720.
Abstract 2721: Real-time and label-free aptasensor of VEGF for cancer diagnostics
Authors: Cho Hansang , Sinha Raghu , Lee Luke P. 
Published: February 2011

Abstract
Early detection of cancer markers is a critical demand for clinical research, particularly in preventing cancer cells from establishing themselves in a metastatic stage. We have developed a stable, sensitive, and selective detection method utilizing a nanoplasmonic ‘aptasensor’ (aptamer-based sensor) of vascular endothelial growth factor (VEGF) for breast cancer diagnostics. Nanoplasmonic aptasensors are prepared by fixing gold nanoplasmonic particles (GNPs) on an amino-silane coated glass slide and coating poly-L-lysine (PLL) on top of GNPs, which provides the GNP with a strong positive-charged surface and a spacing layer avoiding quenching. To accomplish high selectivity, VEGF-specific aptamers are immobilized on the PLL-GNP surface by an electrostatic force. To monitor the event of target binding, the aptamer is conjugated with a fluorescent molecule, Cy-3, which has an absorption peak wavelength (λ = 552 nm) that matches with a localized surface plasmon resonance wavelength of the 80 nm in diameter GNP (λ ≈ 550 nm). In the absence of target VEGF molecules, Cy-3 is spaced in a relevant coupling distance from the GNP surface for the surface plasmon resonance to amplify an incident light and enhance the fluorescent signal of Cy-3. However, once VEGF molecules bind to the aptamer-Cy-3, the conformational change of aptamer occurs, leading to its subsequent detachment from the GNP surface, which results in no signal enhancement and, thus, a loss in the fluorescence intensity. Therefore, the concentration of VEGF molecules can be estimated by measuring the amount of signal decrease in the fluorescence intensity.
With high affinity and specificity, the aptasensor of VEGF requires a small amount of sample (10 μL) for a single detection and incubation time of 1 hour at a room temperature. Also, the aptasensor achieves VEGF detection from 100 nM (2 μg/mL) to 100 pM (2 ng/mL) with a signal decrease up to 50%. In contrast, there is no discernible signal change in a non-specific binding protein solution (100 nM bovine serum albumin) or a strong ionic solution (1.4 M NaCl). Furthermore, the aptasensor is applicable to VEGF detection in human fluids, owing to the biological stability of a fully 2′-O-methyl nucleotide-based aptamer. We can detect VEGF molecules present in saliva and serum of breast cancer patients by observing signal decrease corresponding to the concentration of 100 nM (2 μg/mL) VEGF and a consistent decrease with concentration variation from 1 to 100 % in saline. The aptamer-based nanoplasmonic sensor may be useful for not only basic studies on angiogenesis but also clinical research in evaluating the effectiveness of drug candidates that inhibit VEGF secretion.
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 2721.
Abstract 2726: Salivary biomarkers for breast cancer detection
Authors: Zhang Lei , Xiao Hua , Karlan Scott , Zhou Hui , Feng Ziding , Elashoff David , Akin David , Yan Xinmin , Chia David , Gross Jenny , Karlan Beth , Wong David T. 
Published: February 2011

Abstract
Objectives:
The high burden of breast cancer in women worldwide underscores the unmet potential of biomarker for early detection. New strategies and biomarkers for early detection of breast cancer are sorely needed. In this study, we have conducted a prospective sample collection and retrospective blinded validation (PRoBE design) to evaluate the performance and translational utilities of salivary biomarkers for the non-invasive detection of breast cancer.
Methods:
The Affymetrix HG U133 Plus 2.0 Array and 2D-DIGE were used to profile transcriptomes and proteomes in saliva supernatant from patients with invasive ductal carinoma and matched controls respectively. The discovered transcriptomic and proteomic candidate biomarkers were subjected to a clinical validation using an independent sample set of 31 breast cancer (invasive ductal carcinoma, stages I and II) and 62 healthy controls. Transcriptomic biomarkers were validated by qPCR and proteomic biomarkers were validated by quantitative immuno-blot.
Results:
Significant variations of salivary transcriptomic and proteomic profiles were observed between breast cancer patients and healthy controls. Eleven mRNA biomarker candidates and two protein biomarker candidates were selected for independent validation. Eight mRNA biomarkers and one protein biomarker have been validated for breast cancer detection, yielding ROC-plot AUC values between 0.665 and 0.959. Six biomarkers have known associations with breast cancer.
Conclusion:
Salivary biomarkers possess discriminatory power for the detection of breast cancer, with high specificity and sensitivity. This report provides proof of concept of salivary transcriptomic and proteomic biomarkers for the non-invasive detection of breast cancer. The salivary biomarkers' discriminatory power paves the way for a pivotal clinical validation.
Supported by NIH grant U01DE016275, U01DE017790 and R21CA126733 (DTW).
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 2726.