bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3956: Differential miR-141 expression in primary cultured human breast cancer epithelium (PCHBCEs) and normal adjacent part epithelium (PCHBNEs) correlates with tumor suppressor protein tyrosine phosphataseγ (PTPγ)
Authors: Lin Shu-Hong , Jiang Jinmai , Kuo Chieh-Ti , Ye Weiping , Xu Pingping , Li Hong , Liu Jie-Yu , Zhong Saiyi , Schmittgen Thomas D. , Lin Young C. 
Published: August 2014

Abstract
MicroRNAs (miR) have been shown to be extensively involved in tumorigenesis by post-transcriptional inhibition of oncogenes and/or tumor-suppressor genes. The purpose of our study is to investigate the difference in miRNA expression between cancer epithelium and epithelium from normal breast adjacent tissues. The clinical samples were procured from the Tissue Procurement Program at the Ohio State University Comprehensive Cancer Hospital from breast cancer patients who underwent partial or complete mastectomy. The miRNA profiling for one pair of PCHBCEC and PCHBNEC from the same patient was carried out, and the difference of miRNA expression and potential target genes were further verified by realtime qPCR in 7 pairs of clinical samples. The miRNA profiling showed that 24 miRNAs (let-7a, let-7c, let-7f, let-7g, miR-24, miR-28, miR-29a, miR-29c, miR-30a-5p, miR-30d, miR-92, miR-125a, miR-126*, miR-132, miR-135a, miR-135b, miR-137, miR-141, miR-182, miR-200c, miR-339, miR-365, miR-425-5p, miR-391, p&lt;0.05) are statistically up-regulated in cancer while only 1 miRNA (miR-221) is down-regulated. Based on the magnitude of change and predicted target, we selected miR-141 for further validation. Compared with its normal adjacent counterpart, 4 PCHBCECs had lower miR-141 while 2 were up-regulated and 1 unaltered. Further validation on gene expression of the samples confirmed the negative correlation of miR-141 with its putative target PTPγ. Our comparison of PCHBCECs and PCHBNECs under the same genetic background demonstrated a distinct expression of miRNAs. The dysregulation of miR-141 was shown to result in modulation of the potential tumor-suppressor gene PTPγ which might have an impact on the etiological process of tumor lesion and discriminate cancer epithelial cells from their surrounding normal breast epithelial compartments. Our results implicate that miR-141 might serve as molecular biomarker for therapy of human breast cancer patients. (Supported by NIH R01 Grant ES 015212).
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3956. doi:10.1158/1538-7445.AM2011-3956
Abstract 3961: Sec23A is a regulative target of miR-200c and miR-375 in prostate cancer and influences cell growth in vitro
Authors: Nolte Elke , Szczyrba Jaroslaw , Wach Sven , Jung Volker , Kremmer Elisabeth , Graesser Friedrich , Wullich Bernd 
Published: August 2014

Abstract
Background: Prostate cancer is a leading cause of tumor mortality. In order to identify the underlying mechanisms we have analyzed microRNA (miRNA) expression profiles of primary prostate cancers and non-cancer prostate tissue. Using database analysis, we sought to identify regulatory targets of aberrantly expressed miRNAs and to analyze the physiological consequences of miRNA deregulation using cell culture models.
Material and Methods: We performed comparative miRNA expression profiling using deep sequencing of cDNA libraries and microarray analyses. Potential target genes of deregulated miRNAs were identified by database analysis and subsequently validated in vitro using reporter gene constructs. The impact of identified target genes on cell growth was examined by a standardized wound healing assay.
Results: Two of the miRNAs consistently deregulated in primary prostate cancer samples were miR-200c and miR-375 which were upregulated more than 1.5-fold in deep sequencing and 2.9-fold and 3.9-fold, respectively, in microarray experiments (p&lt;0.001, ANOVA). Independent target prediction databases (including miRanda, Targetscan and PicTar) indicated that the 3’ untranslated region (3’UTR) of the Sec23a gene is a regulative target for both miR-200c and miR-375. This prediction could be confirmed using a luciferase reporter gene construct containing the Sec23a 3’UTR. Co-expression of the reporter gene construct with either miR-200c or miR-375 resulted in a reduction of reporter gene activity while mutation of the miRNA binding sites caused a loss of responsiveness to the respective miRNA. Furthermore we could show that transient expression of miR-200c and miR-375 reduced the amount of Sec23a protein in the HEK293 cell line. A transient knock-down of Sec23a in DU145 prostate cancer cells applying siRNA technique led to acceleration of wound healing and transient overexpression of Sec23a to a deceleration over a time period of 30 hours. In Western blot analyses the majority of primary tissue samples displayed a reduced amount of Sec23a protein.
Conclusion: Our findings demonstrate that the 3’-UTR of Sec23a is one regulative target of miR-200c and miR-375, which are commonly deregulated in prostate cancer. Sec23a is reduced in the majority of tumor samples analyzed and has a vital impact on cell growth as determined by wound healing experiments.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3961. doi:10.1158/1538-7445.AM2011-3961
Abstract 3962: Knocked down miR-221/222 inhibits Akt pathway to suppress malignant glioma cell growth independent of PTEN status
Authors: Zhang Chunzhi , Zhang Anling , Hao Jianwei , Zhang Junxia , Jia Zhifan , Wang Guangxiu , Pu Peiyu , Kang Chunsheng 
Published: August 2014

Abstract
MicroRNAs (miRNAs) are a class of endogenous small noncoding RNAs that regulate gene expression after transcription. By base-pairing with the complementary sites in the 3’untranslated region (3’UTR) of the mRNA, miRNAs control mRNA stability and translation efficiency. Aberrant expression of miRNAs have been shown to be involved in tumorigenesis. MiR-221 and miR-222 were frequently overexpressed in human malignant tumor, including glioblastoma. By Northern Blot assay, we found that miR-221 and miR-222 upregulated in glioma cell lines. In addition, Akt pathway is aberrant activity in glioblastoma. Our work have proved that miR-221/222 promote malignant progression of glioma cell through activation of the Akt pathway. Once Akt activated, it promotes cellular proliferation and invasion ability and reduces cell apoptosis and radiosensitivity. By bioinformation analysis, we found that PTEN is a target of miR-221 and miR-222. By luciferase report assay, we identified that PTEN is a target of miR-221 and miR-222. It is unknown whether miR-221/222 activate Akt pathway dependent of PTEN status or not. We knocked down miR-221/222 with a specific antisense oligonucleotide and found that phenotype of glioblastoma cell changed in vitro in U251 (PTEN mutant) and LN229 (PTEN wild type), including increasing cell proliferation and invasion ability, reducing cell apoptosis and radiosensitivity. By Western Blot, knocked down miR-221/222-cells showed a decreased expression of EGFR, activated Akt, cyclin D, and Bcl-2. Furthermore, knockeddown miR-221/222 considerably decreased tumor growth in U251 cell xenograft model. By immunofluorescence assay, it was found that antisense-miR-221/222 significantly knocked down miR-221 and miR-222 in vivo. By Tunnel assay, antisense-miR-221/222 significantly increased cell apoptosis in vivo. By immunohistochemistry staining, some protein expression reduced, including expression of EGFR, activated Akt, cyclin D, and Bcl-2. Although miR-221 /222 is known to regulate PTEN and downregulation of miR-221/222 led to increased PTEN expression endogenously, the GBM suppressor effect of antisense-miR-221/222 is most likely independent of PTEN regulation because U251 has mutant PTEN. Taken together, our studies provide evidence that miR-221/222 may serve as a novel therapeutic target for malignant gliomas independent of PTEN status.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3962. doi:10.1158/1538-7445.AM2011-3962
Abstract 4056: Fluorescence labeling of different live cell populations and subsequent analysis by flow and imaging cytometry
Authors: Chan Leo L. , Alvarez Mark , Shen Dee , Lin Bo , Patton Wayne F. 
Published: August 2014

Abstract
Relative to conventional antibody-based immunohistological methods employing fixed and permeabilized cells, live cell imaging has presented challenges due to the lack of availability of compatible fluorescent probes. Additionally, instrument-dependent complications arise, such as the requirement that adherent cells be detached into a single cell suspension for flow cytometry, which can impact physiological response. A novel noncovalent fluorescence, long-term labeling technology was employed to stably incorporate green, red, or mixtures of green and red fluorescent dyes into the plasma membranes of different live cell populations. The dyes were well retained for up to 96 hours, faithfully transferred to daughter cells upon mitosis, yet displayed no tendency for nonspecific transfer to adjacent unlabeled cell populations. Since the described dyes do not covalently modify proteins within the cells, normal physiological responses are well preserved. Labeled cell populations were analyzed by conventional flow cytometry as well as with a novel imaging cytometer, the Cellometer Vision, incorporating inexpensive disposable counting chambers. Using a combination of bright-field and fluorescent images, the imaging system automates cell image acquisition and processing using novel counting algorithms for accurate and consistent measurement of cell concentration, fluorescent intensities and percentages of fluorescent positive cells. Results achieved using the imaging cytometer showed excellent correlation with those achieved by flow cytometry, but required 25-fold lower sample per analysis. As many as four cell subpopulations could be determined simultaneously. The described workflow provides a powerful, yet economic, approach applicable to long-term cell viability, cytotoxicity, cell adhesion, cell migration, lineage tracing and cell-cell fusion analysis.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4056. doi:10.1158/1538-7445.AM2011-4056
Abstract 4057: Mono-ubiquitination of Smad2/3 by Smurf2 regulates TGF-β transcriptional response
Authors: Tang Liu-Ya , Yamashita Motozo , Tang Yi , Zhang Ying E. 
Published: August 2014

Abstract
Transforming growth factor-beta (TGF-β) regulates a wide array of biological processes ranging from proliferation to apoptosis, and alterations in its signalling pathway are associated with a variety of human diseases, including cancer. Through Smad proteins, TGF-β signal can be propagated into nuclear transcription machinery. Thus, fine tuning of Smads activity is crucial for TGF-β signal transdction. Smad ubiquitination regulatory factor 2 (Smurf2) is an E3 ubiquitin ligase and previous data shows that Smurf2 negatively regulates TGF-β signaling by inducing polyubiqutination and degradation of Smad proteins. Here, we find a different type of ubiquitination – mono-ubiquitination of Smad2/3 by Smurf2. Smad2/3-Smurf2 interaction and Smad2/3's mono-ubiquitination by Smurf2 are enhanced by TGF-β induced linker phosphorylation. The ubiquitination depends on Smad2/3's PY motif and Smurf2's E3 ligase activity. Cycloheximide treatment assay revealed that this ubiquitination has no effect on Smad2/3 protein stability, but the transcriptional activity is inhibited as shown by both luciferase assay and real-time PCR analysis. Thus, our findings reveal a novel regulatory role of Smurf2 in TGF-β signaling.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4057. doi:10.1158/1538-7445.AM2011-4057
Abstract 4059: Identifying the key domain of CHK1 interaction with MEPE/OF45 and the key ubiquitination site of CHK1
Authors: Chen Guangnan , Yan Dan , Wang Ya 
Published: August 2014

Abstract
Background: Previously, we reported that MEPE/OF45 interacting with CHK1 protects CHK1 from degradation and cells from DNA damage-induced killing (Nucleic Acids Res, 37: 7447-7454 2009; Brit J Cancer 102: 862-866, 2010). To further investigate the nature of the CHK1 stabilization and its effects on cell sensitivity to DNA damage inducers, we designed this study to identify the key domain of CHK1 interaction with MEPE/OF45 and the key ubiquitination site of CHK1.
Methods: GST-MEPE/OF45 was expressed in E. coli BL 21. HA-wild type CHK1 and various CHK1 truncated mutations were expressed in 293FT. GST pull-down assay was used to identify the key domain of CHK1 interacting with MEPE/OF45. To identify the ubiquitination site of CHK1, GFP-CHK1 with different Lys mutations were co-transfected with Ub-expression vector to 293FT, and 48 hours later, whole cell lysate were prepared, Western blot was carried out using a GFP monoclonal antibody to detect ubiquitinaed CHK1. The CHK1 half-life was used to detect the stabilization of CHK1 and a clonogenic assay was used to detect the sensitivity of cells to DNA damage inducers.
Results: The key domain of CHK1 interacting with MEPE/OF45 and the key site of CHK1 ubiquitination were identified. Most importantly, the mutation of CHK1 with the main domain or key site affected CHK1 stabilization and cell sensitivity to DNA damage inducers.
Conclusion: The interaction of CHK1 with MEPE/OF45 plays an important role in preventing CHK1 ubiquitination and; therefore, stabilizes CHK1, which further affects cell sensitivity to DNA damage inducers. Because CHK1 is an essential DNA damage response protein, these results provide valuable information for understanding the role of CHK1 in genomic integrity, which could be used for improving cancer treatment.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4059. doi:10.1158/1538-7445.AM2011-4059
Abstract 4060: Enriching for rrare subpopulations of circulating microvesicles by the depletion of endothelial- and leukocyte-derived microvesicles
Authors: Spetzler David B. , Deng Ta , Tinder Teresa , Kimbrough Jeff , Ellis Phil , Holterman Daniel , Maney Todd , Kuslich Christine D. 
Published: August 2014

Abstract
Endothelial- and leukocyte-derived circulating microvesicles (cMV) represent a majority of the cMV present in the blood. The purpose of this study was to determine if depletion of these more common cMV populations is possible, allowing for the enrichment and analysis of the remaining subpopulations of cMV. Circulating microvesicles were isolated from the blood plasma of breast cancer patients. Magnetic beads conjugated to antibodies specific for either CD31 or CD45 were used to deplete the isolated endothelial- and leukocyte-derived cMV. The remaining cMV population was characterized with a multiplexed immunoassay against a panel of 20 different antigens. In addition to the expected depletion of the biomarker used to remove the targeted cMV in the remaining cMV population, a concomitant depletion of associated biomarkers was observed. In the case of the cMV population in which CD31 had been used to remove endothelial-derived cMV, for instance, there was a significant concomitant depletion of DLL4, a highly associated endothelial marker. This was not the case with general epithelial microvesicle markers, such as CD9, which were still very much present in the cMV population remaining after depletion. These findings suggest a potential method for a more complete characterization of biomarker profiles associated with cMV derived from disease-associated cells and provide the foundation for a novel cMV-based strategy for disease detection through a noninvasive blood test.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4060. doi:10.1158/1538-7445.AM2011-4060
Abstract 4134: Exploration for possible predictive biomarkers for individual response to mFOLFOX6 in colorectal cancer patients
Authors: Suzuki Okihide , Murata Kohei , Fukunaga Mutsumi , Takemoto Hiroyoshi , Ohue Masayuki , Ikeda Rikuko , Wada Satoru , Eguchi Hidetaka , Tomita Naohiro , Watanabe Masahiko , Ishida Hideyuki , Nishiyama Masahiko 
Published: August 2014

Abstract
Background: Incorporation of molecular targeting agents such as bevacizumab, cetuximub, and panitumumab, into cytotoxic chemotherapeutic regimens has increased survival in colorectal cancer patients, resulted in provision of a promising means of individualization of these therapies. Nonetheless, the clinical efficacy and toxicity of basic regimens, such as FOLFIRI and FOLFOX, remain unpredictable. Thus, identification of molecular indicators of individual response to the basic regimens is urgently required. Methods: This prospective study was conducted to determine potent predictive markers of individual response to mFOLFOX6, in terms of efficacy and toxicity. Chemonaive patients with stage IV colorectal cancer were eligible for this trial after a palliative operation, and received mFOLFOX6 treatment. Along with a search for KRAS gene somatic mutations, analyses of mRNA expression levels in cancerous portions were conducted for 10 candidate marker genes, namely DPYD, TYMS, ERCC1, ERCC2, XRCC1, GSTP1, EGFR, VEGF, TYMP and TNFRSF1B, by means of real-time RT-PCR. In addition, some acknowledged polymorphisms of DPYD, TYMS, ERCC1, ERCC2, XRCC1, GSTP1 and EGFR genes were also determined. Results: Fifty pts received a total of 424 treatment cycles. Overall response rate was 60.4% (1CR and 28 PRs), and median overall and progression survival were 930 days and 234 days, respectively. The most common grade 3/4 toxicity was neutropenia (40%), followed by anorexia (6%), anemia (4%), leucopenia (4%), etc. The therapeutic response was not related to KRAS somatic mutational status, and no genotype-toxicity association was observed, thus far tested. On the other hand, ERCC1, ERCC2, and TYMS gene expression levels were significantly higher in patients with PD than in those with CR/PR/SD (P=0.017, 0.028, and 0.032, respectively). The expression levels of TYMS were highly correlative with best tumor response rate (R=0.414, P=0.004). Overall survival was significantly better in patients with low ERCC2 expression (P=0.039). Furthermore, combined expression levels of ERCC1, ERCC2, and TYMS were found to be associated with PFS: Those patients with high expression levels of all of these 3 genes showed poorer survival than the others (P=0.0029). Conclusions: Expression levels of ERCC1, ERCC2, and TYMS genes in tumor may be predictable for individual response to mFOLFOX6. A large-scale prospective study to validate their predictive values is now under planning.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4134. doi:10.1158/1538-7445.AM2011-4134
Abstract 4156: Detection of lung cancer from plasma using the biosignature of circulating microvesicles
Authors: Tasinato Andrea , Hassaine Gherici , Ristorcelli Elodie , Chen Wang-Juh , Kuslich Christine D. 
Published: August 2014

Abstract
Circulating microvesicles (cMV) are cell-derived structures that are abundantly present in the blood. Tumor cells produce large quantities of cMV, and their abundance has been shown to correlate with tumor invasiveness and resistance to therapy. This study attempts to understand the origin, composition, and potential clinical utility of cMV, analyzing their protein composition in patients with non-small cell lung cancer (NSCLC) and identifying a cMV-based biosignature that can be used to predict the presence of tumors from a blood sample.
cMV were isolated from plasma samples that were obtained from an initial cohort of 65 patients with NSCLC and from 46 control samples. A discovery panel of 63 specific biomarkers was used to develop an assay with high specificity and sensitivity based on 4 cMV surface proteins. Using a novel cMV-based multiplexed analysis platform we optimized a threshold level for 4 significant cMV subpopulations to effectively distinguish lung cancer patients from controls.
The multiplex assay included 1 general cMV marker (CD81) and 3 lung cancer-associated biomarkers: the lung epithelial C-type lectin members SPD and SPA, plus the sialoprotein osteopontin. The decision tree analysis, based on specific cut offs, showed a sensitivity of 81% and a specificity of 90%.
These results provide initial evidence that the identification of biosignatures in distinct subpopulations of cMV may offer a powerful blood-based approach for the detection and monitoring of specific disease states, such as NSCLC.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4156. doi:10.1158/1538-7445.AM2011-4156