bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4579: Effect of pre-adipocyte and differentiated adipocyte conditioned media on galectin-3 expression in mouse intestinal epithelial cells
Authors: Kim Jong-Sang , Lim Ji-Sun , Garcia Annalene , Seo Ji-Yeon , Park Jia , Nam Dae Hwan , Sung Mi Kyung , Seo Young-Rok 
Published: February 2011

Abstract
We hypothesized that obesity cause inflammation which, in turn, promotes colon carcinogenesis. To test the hypothesis mouse intestinal IEC-6 cells were incubated in the presence of pre-conditioned media collected from culturing LPS-stimulated macrophage, followed by comparing protein expression profile with control cells. Our proteomic study revealed 45 differentially regulated proteins in mouse intestinal IEC-6 cells under state of inflammation. Among others, galectin-3, a beta-galactoside-binding protein, was up-regulated. Galectin-3 has been reported to play anti-apoptotic role in tumor progression and metastasis, and its over-expression has been observed in several types of cancer such as colon cancer. In order to determine relationship between obesity-induced inflammation and colon carcinogenesis, conditioned media (CM) from LPS-stimulated 3T3-L1 pre-adipocytes or differentiated adipocytes were utilized to culture IEC-6 cells. The CM was analyzed for levels of nitric oxide (NO), TNF-α and IL-6, and IEC lysates were analyzed for galectin-3 expression by western blot. Results showed that enhanced generation of NO, TNF-α and IL-6 resulted in upregulation of galectin-3 with respect to incubation time. Also, cytokine concentration was correlated to level of galectin-3, as verified by incubating IEC with increasing concentration of TNF-α or IL-6. These results indicate involvement of pro-inflammatory mediators from pre-adipocytes and adipocytes in colon carcinogenesis, in which galectin-3 may play a crucial role.
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 4579.
Abstract 4580: Detection of early-stage pancreatic cancer by mass spectrometry: a multi-institutional validation study
Authors: Honda Kazufumi , Ono Masaya , Okusaka Takuji , Kosuge Tomoo , Felix Klaus , Nakamori Shoji , Sata Naohiro , Nagai Hideo , Ioka Tatsuya , Tsuchida Akihiko , Shimahara Masashi , Yasunami Yoichi , Hirohashi Setsuo , Büchler Markus W. , Yamada Tesshi 
Published: February 2011

Abstract
Background Invasive ductal carcinoma of the pancreas is one of the worst prognoses among common human malignancies. The poor outcome of pancreatic cancer patients seems to be attributable to difficulty in early detection.
Methods Mass spectra of peptide were obtained from a total of 1314 plasma/serum samples, consisting 4 cohorts (namely Cohorts 1-4) obtained from 7 medical institutions participating in the “Third-Term Comprehensive Control Research for Cancer” conducted by the Ministry of Health, Labor and Welfare of Japan and one institute in Germany using orthogonal matrix assisted laser/desorption ionization time of flight (oMALDI-qQTOF-MS). The amino acid sequences and posttranslational modifications of marker proteins were identified by tandem mass spectrometry (MS/MS).
Results Plasma apolipoprotein AII heterodimer (ApoAII-2; 17252 m/z), unglycosylated apolipoprotein CIII (ApoCIII-0; 8766 m/z), and their summated value differed between 112 pancreatic cancer patients and 103 healthy controls (Cohort 1) with high statistical significance [P = 1.36 × 10−21, P = 4.35 × 10−14, and P = 1.83 × 10−24 (Mann-Whitney U-test)] and area-under-curve (AUC) values of 0.877, 0.798, and 0.903, respectively. The significance was further verified in 2 retrospective cohorts [Cohort 2 (n = 103) and Cohort 3 (n = 163)] and a hospital-based prospective cohort [Cohort 4 (n = 833)]. Patients with invasive ductal adenocarcinoma of the pancreas, including those with stage-I disease, could be discriminated from healthy controls with the sensitivity of 97.6% (243/249) and specificity of 95.3% (122/128) by the combination of DUPAN-2, CA19-9, ApoAII-2, and ApoCIII-0.
Interpretation Although the reduction of the 2 modified forms of plasma/serum apolipoproteins was not pancreatic cancer-specific, the accurate measurement of plasma/serum proteins had a high capability in detecting pancreatic cancer. This test is essentially safe and might be applicable to the primary mass screening of pancreatic cancer in asymptomatic populations.
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 4580.
Abstract 4583: Multiplex mass spectrometric immunoassays
Authors: Nedelkov Dobrin 
Published: February 2011

Abstract
Targeted proteomics approaches can facilitate the study of the human protein variations and their association with cancer. In Mass Spectrometric Immunoassays, affinity interactions are utilized to selectively retrieve specific proteins from complex biological fluids, and are followed by mass spectrometric analysis of the targeted proteins. These assays are similar to traditional enzyme immunoassays in that they use antibodies as reagents for affinity retrieval of specific proteins, but instead of enzymatic reaction for indirect protein detection, mass spectrometric analysis is used for direct identification of the proteins and their modifications. This rather straightforward concept is realized through affinity pipette devices that enable high-throughput integration and assaying of thousands of samples per day.
Described here is the development of several multiplex mass spectrometric immunoassays toward cancer-related proteins. The project is part of the NCI's Clinical Proteomic Technologies for Cancer initiative to assess and apply proteomic technologies and data resources to solve critical problems in cancer research. Antibodies to several proteins (including one that serves as an internal reference standard - IRS) were immobilized in various ratios in the affinity pipettes, and following analysis of human plasma spiked with constant amount of the IRS, the ratio of the peaks representing the proteins were monitored to determine the optimal ratio of antibodies in the affinity pipettes. Detailed assaying protocols were developed and optimized, including pre-and post-sample rinses, and elution matrix composition. Increasingly-diluted plasma samples were analyzed and a decrease of the protein signals in reference to the IRS signal was observed, indicating non-saturating conditions for the assays. The assays were tested with numerous human plasma samples, and the intra- and inter-assay CVs were determined to be &lt;10%. The results obtained with the MSIA assay were also compared to commercially available ELISA assays.
The introduction of mass spectrometric-based assays and platforms into clinical laboratories will facilitate the ongoing biomarker discovery, evaluation, and validation efforts. The multiplex mass spectrometric immunoassays are idealy suited for analysis of protein biomarkers exhibiting post-translational modifications because this protein microheterogeneity can only be detected via mass spectrometry. It is important to note that the assays target intact/native proteins (in contrast to MRM-based peptide assays), and in such are capable of detecting any and all PTMs, without a priori knowledge of their existence.
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 4583.
Abstract 4587: Quantitative analysis of IGF-1R signaling pathway activation in FFPE tissue
Authors: Krizman David , Hembrough Todd , Hiedbrink-Thompson Jenny , Thyparambil Sheeno , Burrows Jon , Darfler Marlene , Prakash Amol , Krastins Bryan , Sarracino David , Rezai Taha , Athanas Michael , Lopez Mary 
Published: February 2011

Abstract
Multiple Ewing's sarcoma clinical trials targeting insulin-like growth factor receptor (IGF-1R) with IGF-1R small molecules and antibody inhibitors are currently underway. The IGF-1R signal pathway is commonly activated in many cancer types so this targeted therapeutic approach holds great promise for not only treating Ewing's sarcoma but other types of cancers as well. There is currently no good method to quantitatively measure the IGF-1R protein, its phosphorylation status, or the immediate downstream signal pathway target IRS-1. Being able to quantify these proteins and determine their phosphorylation status directly in patient tissue biopsies could help identify and select patients most likely to benefit from anti-IGF-1R therapies. We have developed an approach to determine both absolute IGF-1R and IRS-1 levels and the phosphorylation status of both proteins directly in formalin-fixed paraffin-embedded (FFPE) patient tissue. This approach is based on the Liquid Tissue®-SRM technology platform, a combination of tissue microdissection, Liquid Tissue® processing which turns dissected tissue to a complete solubilized tryptic digest, and mass spectrometry-based selected reaction monitoring (SRM). This approach enables relative and absolute quantification of proteins and their phosphorylation status directly in formalin fixed tissue. This approach was used to measure the IGF-1R protein and its phosphorylation sites pY1131, pY1135, and pY1136 as well as the IRS-1 protein and its phosphorylation sites pY612 and pY896 in formalin fixed tissue culture cells and xenograft tumors. Results indicate the potential to detect and quantify the IGF-1R and IRS-1 proteins and to monitor their phosphorylation status directly in FFPE tumor tissue which could be used to identify and stratify patients most likely to benefit from anti-IGF-1R therapies.
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 4587.
Abstract 459: Caveolin-1 mediated signaling and proliferation is regulated through the oxygen sensing pathway
Authors: Xu Chao Ying , Wang Yi , Roche Olga , Ohh Michael 
Published: February 2011

Abstract
Caveolin-1 (Cav-1) is a multifunctional scaffolding protein with multiple binding partners that is associated with cell surface caveolae and the regulation of lipid raft domains. Cav-1 regulates multiple cancer-associated processes including cellular transformation, tumour growth, cell death and survival, and multidrug resistance. A high expression of Cav-1 is found in various cancer , including clear cell renal cell carcinoma (CCRCC), correlating with large, high grade tumors and poor prognosis. However, the role of Cav-1 in the oncogenesis of CCRCC remains unclear, and its role as either an oncogene or tumour suppressor is still under debate. Here, we investigated the mechanism of Cav-1 in controlling cell proliferation using in vitro and in vivo assays. We report that Cav-1 is highly expressed in tumour compared to normal tissue in patients with CCRCC, thus Cav-1 likely functions as an oncogene. Upon knock-down of Cav-1, tumour cells (786-VHL-/-) showed a decrease in proliferation. We showed that Cav-1 is associating with EGFR, a receptor tyrosine kinase, to activate the Ras-RAF-MEK-ERK kinase pathway and induce cell proliferation. We found that Cav-1 binds to EGFR in immunoprecipitation. The Ras pathway is upregulated in cells overexpressing Cav-1 after EGF stimulation. Upon knockdown of Cav-1, the level of p-ERK, p-MEK, and p-c-Raf decreased, as well as the rate of cell proliferation. Upstream of the Ras pathway, we report that Cav-1 appears to be under the control of hypoxia-inducible factor (HIF). Under hypoxia, VHL (the principal negative regulator of HIF) is downregulated, thus leading to upregulation of HIF. We showed that under hypoxia, Cav-1 increased in 786-VHL, Hela and A431 cell lines. Similarly, cells with VHL loss (786-VHL-/-), thus high HIF level, had high levels of Cav-1. We are currently examining the role of Cav-1 in tumour growth by injecting SCID mice with either 786-VHL-/-, or Cav-1 knock-down 786-VHL-/- tumour cells. Since Cav-1 level seems to be under the control of HIF in renal, cervical and squamous carcinoma cell lines, it suggests that the oxygen sensing pathway plays a general role in controlling cell proliferation through upregulating Cav-1. Cav-1 should be considered as a target for drug design as the knock down of Cav-1 decreased cell proliferation and could potentially decrease tumour growth.
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 459.
Abstract 4591: Estrogen receptor beta controls a wide variety of functions in breast cancer cells
Authors: Cekanova Maria , Woraratphoka Jirayus , Sukhthankar Mugdha , Dharmawardhana S , Siriwardana Nalin , Moustaid-Moussa Naima , Strom Anders , Baek Seung J. , Wong Q , Zou M , Donnell Robert , Wimalasena Jay 
Published: February 2011

Abstract
The role of ER alpha as the mediator of estrogen action in breast cancer (BC) cells is well known, however, the role of ER beta is controversial. The majority of BC expressing ER alpha also express ER beta. Some clinical studies suggest that ER beta expression is a good prognosticator; however, in others, especially where expression levels of ER beta &gt; ER alpha, ER beta may be associated with more advanced BC. Previously, we showed that ER beta may regulate a number of signaling pathways (AACR, 2008). Our recent work demonstrates that ER beta has even wider actions in MCF7 and T47D cells stably overexpressing ER beta. Thus, ER beta decreased phospho (p)/ total Stat1, p Stat3, p Stat5/total Stat5. Expression of the oncogenic miR cluster 17-92 was highly reduced (p&lt;0.01) when ER beta overexpressing vs. control cells were compared. ER beta also decreased PPAR gamma, superoxide dismutase, as well as the concentration of the mRNA processing factor, RBM5, and the activated form of AMPK. All these changes were significant (p&lt;0.05) in triplicate measurements. While ER beta overexpression increased migration through Matrigel, it significantly decreased CXCR4 expression (p&lt;0.05, n=4). All of these effects of ER beta were estrogen-independent. On the other hand, estrogen increased CXCR4 expression in MCF7 cells significantly. In our study, we analyzed the expression of ER beta and BAD using tissue microarrays with normal and neoplastic breast tissue. Our immunohistochemical (IHC) data revealed a significant increase of ER beta cytoplasmic staining in neoplastic cells compared to normal cells (p&lt;0.001). In addition, the nuclear expression of ER beta was significantly decreased in neoplastic cells compared to normal cells (p&lt;0.01). From our correlation analysis of IHC data obtained from tissue microarray, we identified that cytoplasmic expression of BAD was negatively correlated with cytoplasmic expression of ER beta. In addition, the nuclear expression of BAD was significantly positively correlated with nuclear ER beta.
These results suggest that ER beta has a wide variety of actions in BC cells, as predicted by the regulation of 62 miRs (p&lt;0.01) as determined by Exiqon Inc. Taken together our results suggest that ER beta may have a multitude of actions in breast cancer and that the ratio of ER alpha/beta may determine the prognostic outcome.
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 4591.
Abstract 4612: Analysis of transcriptional regulation by progesterone receptor in ER-positive breast cancer cells by ChIP-Seq
Authors: Yang Fan , Bilke Sven , Pineda Marbin , Walker Bob , Zhu Jack , Meltzer Paul 
Published: February 2011

Abstract
Breast cancer is among the most common malignancies in women. It is well established that progesterone receptor (PR) is a major determinant in the development and progression of breast tumor, as well as in diagnosis and treatment of breast cancer. However, it is still not fully understood how PR affects downstream signaling pathways and cross talks with other intracellular signal transduction pathways. To better define the role played by PR in estrogen receptor (ER)-dependent gene regulation, we have used chromatin immunoprecipitation coupled with next generation sequencing (ChIP-Seq) to examine the in vivo DNA binding by PR. Whole-Genome ChIP-Seq analysis has identified large number of PR binding sites in T-47D breast cancer cells, which are ER-positive and PR-positive. PR binding sites were detected in proximal promoter regions as well as distal regions, with most located in non-proximal regions. The comparison of PR and ER binding sites revealed that a significant of portion of the PR binding sites overlaps with the ER binding sites, indicating a close relationship between PR and ER in ER-dependent gene regulation. The PR motifs were found to be among the most enriched sequence motifs within the PR binding sites. Further analysis using the chromosome conformation capture assay revealed that progesterone induces changes to the chromatin structures of known PR-regulated genes. The ChIP-Seq data, together with the expression profiling and other results, indicate that PR may cooperate with ER and play an important role in ER-dependent gene regulation, mammary epithelial differentiation and breast cancer development.
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 4612.
Abstract 4615: Mechanism of inhibition of MMTV-neu, -wnt1 and -Myc induced mammary oncogenesis by RARα agonist Am580
Authors: Farias Eduardo F. , Lu Yongke , Bertran Silvina , Garcia Avalon , Jones Alexis , Samuels Tracey-Ann 
Published: February 2011

Abstract
We tested whether specific activation of a single retinoic acid receptor, RAR alpha (RARα), without direct activation of RAR beta (RARβ) and gamma (RARγ), will inhibit mammary tumor oncogenesis in murine models relevant to human cancer. Fifty MMTV-neu (neu), 30 MMTV-myc (myc), and 50 MMTV-wnt1 (wnt1) transgenic mice were treated with 0.3mg/Kg/day RARα agonist, (retinobenzoic acid, Am580), added to the diet for 40 (neu), 50 (myc) and 35 weeks (wnt1), respectively. Different responses to the treatment were observed in the 3 mouse models, the wnt1 the more responsive, the neu with a mild response and a subset of the myc refractory to the treatment. Among the anti-tumor effects observed in the wnt1 and neu models was the inhibition of epithelial hyperplasia, a significant increase (p&lt;0.05) in tumor-free survival, a reduction in tumor incidence and in growth of established tumors and lung metastasis dissemination (neu model, p&lt;0.001). In both models the mechanisms responsible for these effects involved inhibition of proliferation and survival pathways, and induction of apoptosis. The stronger wnt1 response to Am580 also induced differentiation, both in vivo, and in 3D cultures. In these tumors Am580 inhibited the wnt-pathway, measured by loss of nuclear b-catenin, suggesting partial oncogene-dependence of therapy. Am580 treatment increased RARβ and lowered the level of RARγ, an isotype whose expression we linked to tumor proliferation. Interestingly, the myc mice showed a more complex response in which no significant effect was observed on tumor development but a clear reduction on tumor growth (p&lt;0.001) and lung metastasis dissemination (p&lt;0.01) in a subset of mice (63%). Immunohistological analysis showed scattered necrotic foci, induction of Cyp26A1, E-cadherin, CRBP1 and p27, known RARα response genes, suggesting that cell death, growth inhibition and differentiation are involved in the anti-tumor effect of Am580 in this model. As in the neu and wnt1 models, RARγ was overexpressed in the myc tumors, genetic and pharmacological approaches inhibiting RARγ expression and activity resulted in growth inhibition and differentiation in vitro (2D and 3D cultures). Finally, forced expression of CRBP1 in myc tumor derived cells inhibited tumor growth in xenografts (60%, p&lt;0.05) suggesting that the induction of CRBP1 expression by Am580 can be, in part, responsible for the anti-tumor effect. The anti-cancer effect of RARα, together with the newly discovered pro-proliferative role of RARγ, suggests that specific activation of RARα and inhibition of RARγ might be effective in breast cancer 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 4615.
Abstract 4616: XR5944 interacts with diverse EREs differently, which may influence ER inhibition by XR5944
Authors: Zhang Zhenjiang , Shu Feng-jue , Chen Xi , Carver Megan , Kallen Caleb , Sidell Neil , Yang Danzhou 
Published: February 2011

Abstract
Since the discovery of the estrogen receptor (ER) as the modulator of estrogen action, ER has evolved to be one of the most effective targets for breast cancer therapy. However, the current ER-targeting therapies have a main limitation related to resistance. Our previous studies have shown that XR5944, a DNA bis-intercalator with potent anticancer activity, is capable of inhibiting the ER activity by its ability to specifically block the binding of ER to its consensus DNA sequence ERE compared to other transcription factor-target DNA interactions. This novel mechanism of ER inhibition by a DNA bis-intercalator may be useful in overcoming resistance to current antiestrogens. The consensus ERE is an inverted repeat comprised of two ERE half-sites separated by three bases: AGGTCAnnnTGACCT where nnn is known as the tri-nucleotide spacer. ER can also bind to non-consensus EREs, i.e. natural EREs that differ from the consensus ERE by one or more base pairs but nevertheless serve as potent transactivation sites for estrogen target genes Therefore, to further understand the mechanism and specificity of the inhibition of ER by XR5944, we tested the specificity of XR5944 to block interactions with consensus and non-consensus ERE sequences by 1D 1H NMR titration studies. Our NMR studies of XR5944-DNA interactions suggested that the consensus ERE spacer plays a significant role in determining the binding characteristics of the complex. Of those tested, the CGG/GCC and AGG/CCT spacers showed the best binding, while the CGT/ACG and TTT/AAA spacers showed the worst binding with XR5944. The binding stoichiometry of XR5944 with ERE sequences appears to be 2:1, which explains why the spacer region can affect the drug-DNA binding. To validate our NMR results, we have also conducted functional studies by using reporter constructs containing EREs with tri-nucleotide spacers CGG and TTT. Results of luciferase reporter assays in MCF-7 cells indicated that XR5944 was significantly more potent in inhibiting the activity of reporters containing ERE-CGG than those containing ERE-TTT, consistent with our NMR results. Together with our NMR studies, these reporter gene experiments indicate that the tri-nucleotide spacer plays a significant role in the binding characteristics and functional consequence of XR5944 interactions with the consensus ERE.
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 4616.