bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4006: Biochemical characteristics of functional domain of KITENIN: Its implication in AP-1 signal activation and colorectal cancer cell motility
Authors: Bae Jeong A , Sun Eun Gene , Park So Yeon , Kim Hangun , Kim Kyung Keun 
Published: August 2012

Abstract
Background & aims: KITENIN (KAI1 C-terminal interacting tetraspanin) is not only a membrane-associated protein but also a functional protein promoting the invasiveness of colorectal cancer (CRC) cells. Recent studies about the mechanism of KITENIN function in the CRC cells indicate that KITENIN plays a pivotal role in tumor initiation and promotion through activating the activator protein-1 (AP-1) signaling. However, the contribution of each functional domain of KITENIN to AP-1 signaling is still unknown and the biochemical properties of C-terminal region of KITENIN were investigated. Methods: To compare the biochemical and functional properties of KITENIN deletion mutants with wild-type KITENIN, the various mutant constructs of C-terminal region (238-512 aa) were generated and being expressed in the 293T and Caco2 cells. AP-1 activity was measured via AP-1 luciferase reporter assays, and the cellular phenotypes of these cells expressing KITENIN mutants were examined via in vitro cell invasion and wound-healing assays. Results: KITENIN mutant with deletion of C-terminal 63 amino acids (DCKIT) markedly increased the AP-1 activity in contrast to other deletion mutants and wild-type KITENIN. Also, DCKIT increased the degradation of Dvl2 and stability of c-Jun, which are responsible for AP-1 activation by KITENIN. The C-terminal fragment of KITENIN (KICD, roughly 30 kDa) was generated and preferentially translocated to the nucleus after PMA treatment, but its functional significance is under investigation. A mutant construct of 408ALRA411-KITENIN was obtained by mutation of YXXΨ motifs, which directs the clathrin-dependent endocytosis of membrane-spanning proteins. This mutation did not influence the AP-1 activity, cell invasiveness, and cell migration by KITENIN but disrupted the interaction of KITENIN with Dvl2. Moreover, co-expression of this mutant with ErbB4 exhibited more attenuated AP-1 activity than that of forced expression of wild-type KITENIN and ErbB4. These results indicate that endocytosis of KITENIN and subsequent interaction with Dvl2 is essential to activating AP-1 signal and modulating cell motility by KITENIN. Conclusion: These biochemical properties of deletion mutants of KITENIN suggest that the C-terminal region of KITENIN is an important component for modulating AP-1 signal and CRC cell motility by KITENIN.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4006. doi:1538-7445.AM2012-4006
Abstract 4009: Hypoxia potentiates Runx2-mediated increased Bcl-2 expression in prostate cancer resulting in apoptosis resistance
Authors: Browne Gillian , McCook Heather , Ming Louise , Stein Janet L. , Stein Gary S. , Lian Jane B. , McKeown Stephanie R. , Worthington Jenny 
Published: August 2012

Abstract
Hypoxia is known to act as a selective pressure in the tumor microenvironment, often resulting in sub-populations of cells that have a propensity for more aggressive behaviour. More recently, Runx2 (an osteogenic transcription factor) has been associated with tumor growth, invasion and metastasis. Previous results in our laboratory have indicated that hypoxia selects for more invasive, apoptosis-resistant prostate cancer cells. Microarray analysis revealed that both Runx2 and Bcl-2 (an anti-apoptotic protein) were up-regulated in the hypoxia-selected cells (Butterworth et al, 2008). Bcl-2 is known to be up-regulated in various malignancies, and is associated with more aggressive disease and resistance to chemotherapy. Subsequent analysis of the Bcl-2 promoter revealed two binding sites for Runx2. In the current study we confirmed regulation of Bcl-2 by Runx2 though ChIP binding and reporter assays in LNCaP prostate cancer cells. Runx2 over-expressing LNCaP cells showed increased cell viability which was inhibited by Runx2 siRNA knockdown. A variety of assays demonstrated that Runx2 over-expressing cells were more resistant to apoptosis than controls. We further addressed the effect of hypoxia on Runx2 expression and the potential role Runx2 might play in the regulation of Bcl-2 expression and thus apoptosis in prostate cancer. When LNCaP cells were exposed to hypoxia, both Runx2 and Bcl-2 mRNA and protein levels were significantly increased over oxic controls. In a separate in vivo study, treatment of LNCaP tumors grown on the backs of immuno-compromised mice with the anti-androgen bicalutamide resulted in a profound decrease in tumor oxygenation during the first 14 days. Over the following 14 days, oxygen levels gradually increased. Further investigation of the excised tumors showed an increase in Runx2 mRNA levels from day 7, which was further confirmed by western and immunohistochemical analyses. In summary, this study shows that hypoxia promotes over-expression of Runx2 in prostate cancer cells. The cells exhibit a survival advantage through up-regulation of Bcl-2 leading to apoptosis resistance. These results, taken together with the in vivo data, suggest that Runx2 expression (either through microenvironmental pressures or ectopic expression) promotes progression to a more apoptosis-resistant and thus malignant phenotype, highlighting Runx2 as a potential therapeutic target in prostate cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4009. doi:1538-7445.AM2012-4009
Abstract 4012: Pyruvate kinase M2 promotes the growth and migration of gastric cancer cell via nuclear factor-kappa B and its target genes
Authors: Kwon Oh-Hyung , Kang Tae-Wook , Kim Jeong-Hwan , Kim Mirang , Noh Seung-Moo , Song Kyu-Sang , Yoo Hyang-Sook , Kim Woo-Ho , Xie Zhi , Pocalyko David , Kim Seon-Young , Kim Yong Sung 
Published: August 2012

Abstract
Tumor cells take up more glucose than normal cells and convert large amounts of glucose to lactate even in the presence of oxygen. This phenomenon is known as aerobic glycolysis, and is important for cell growth and metastasis. PKM2 is an isoenzyme of the glycolytic enzyme pyruvate kinase, and promotes aerobic glycolysis. Here, we describe an important role for PKM2 in regulating the survival and migration of gastric cancer cells. Using qRT-PCR and tissue microarray, we showed that PKM2 was overexpressed in gastric tumor tissues compared to normal tissues. Furthermore, PKM2 expression levels were inversely correlated with gastric cancer patient survival. Using shRNA-mediated knockdown of PKM2, we showed that PKM2 regulated apoptosis-related gene, Bcl-xL, at transcriptional levels, and affected cell survival. PKM2 also regulated migration-related gene, MMP-9, at transcriptional levels, and affected cell migration. The effect of PKM2 on cell death and migration was mediated by the p65 stability resulting in binding activity on its target gene promoter. These results provide new insight that PKM2 is an up-stream molecule regulating p65 function and that PKM2 may be a useful target for gastric cancer therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4012. doi:1538-7445.AM2012-4012
Abstract 4018: DNA methylation controls CHST11 gene expression in breast cancer
Authors: Herman Damir , Leakey Tatiana I. , Jousheghany Fariba , yao-Borengasser Aiwei , Cooney Craig A. , Kieber-Emmons Thomas , Monzavi-Karbassi Behjatolah 
Published: August 2012

Abstract
Introduction: We have previously demonstrated that chondroitin sulfates (CS) play a crucial role in metastatic potential of breast cancer cells. We have shown that the expression level of carbohydrate (chondroitin 4) sulfotransferase-11 (CHST11) correlates with CS-A expression and aggressiveness of human breast cancer cells. The current study was mainly performed to determine whether the expression of CHST11 gene in breast cancer cells is controlled by DNA methylation. Methods: Quantitative real-time PCR was used to detect the expression of CHST11 in human breast cancer cell lines. MCF7 cells were treated with 5-aza-2′-deoxycytidine (5AzadC) for DNA demethylation and expression analysis. Methylation in MCF7, MDA-MB-231 cells and patient breast cancer specimens was analyzed using bisulfite genomic sequencing (BGS). Reduced representation bisulfite sequencing (RRBS) was performed in the CpG island of CHST11 with a minimum coverage of 10. Digital gene expression was used to estimate the number of transcribed copies of the CHST11 gene from the second-generation mRNA-sequencing data. Results: Treatment of MCF7 cells with 5-AzadC increased the expression of CHST11 and its product CS-A in a dose-dependent manner. We observed very low levels of DNA methylation in a CpG island of CHST11 in estrogen receptor-negative basal-like cell lines MDA-MB-231, MDA-MB-468, and BT-20, but very high levels in the same region in estrogen receptor-positive luminal MCF7, ZR-75-1, and T47-D cells. We found that the CpG island can also be hypomethylated in triple negative (negative for estrogen and progesterone receptors and Her2/neu) patient breast cancer tissues. Conclusions: The data suggest that DNA hypomethylation significantly contributes to CHST11 and surface CS-A expression in aggressive breast cancer cells. The data suggest that therapeutic use of DNA demethylating agents may promote more aggressive forms of breast cancer.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4018. doi:1538-7445.AM2012-4018
Abstract 4037: Downregulation of ZNF516 and INTS1 is related with promoter methylation in cervical uterine cancer cell lines
Authors: Ili Carmen G. , Brebi Priscilla , Andana Alejandra , Leal Pamela , Garcia Patricia , Guerrero-Preston Rafael , Roa Juan C. 
Published: August 2012

Abstract
Introduction: One of the most common neoplasia in developing countries is cervical uterine carcinoma (CC). HPV infection is recognized as the major risk factor for carcinogenesis of cervix. However, not all infected women will develop a cervical cancer. Several studies have proposed DNA methylation as early biomarker in CC. A higher promoter methylation is associated with lower gene expression. Aim: To evaluate the association between DNA methylation and gene expression of ZNF516 and INTS1 in cervical cancer cell lines. Materials and Methods: DNA isolated from 12 normal and 7 cervical cancer tissue samples was enriched by Methylated DNA Immunoprecipitation (MeDIP) and hybridized to Nimblegen 385K CpG Islands plus Promoter arrays (MeDIP-chip). Bioinformatics strategies were used for background correction, array normalization and data analysis of differentially methylated genomic regions by Methylation Specific PCR in a cohort of 221 normal, premalignant, and cervical cancer samples. Relative expression analysis was performed in a normal cervical epithelium cell line (ECT1 E6/E7) and three cervical cancer cell lines (C-4I, SiHa and C-33A) using real time PCR. MSP was also used to examine promoter methylation in normal cervical and cervical cancer cells lines. Results: ZNF516 and INTS1 were differential methylated in the promoter region of normal and cervical cancer samples in the MeDIP-chip discovery project. The methylation frequency was 100% in cervical cancer cell lines. Relative expression of ZNF516 was significantly lower in C-4I and SiHa (p &lt;0.05) in relation with the normal cell line. C-33A showed elevated promoter methylation and a low expression of ZNF516, however, this result was not significant. INTS1 expression was significantly lower in C-33A and SiHa (p &lt;0.05) in relation with the normal cell line. C-4I showed a high promoter methylation and a low expression of INTS1, however, this result was not significant. Conclusions: We have identified ZNF516 and INTS1 as potentials biomarkers for cervical cancer. These genes may be a useful tool for diagnosis and clinical management of cervical cancer. These results need to be confirmed in clinical samples, from an independent laboratory before the gene panel is examined in a Phase II Biomarker Development Trial. Work sponsored by CORFO Project 09CN14-5960 and Postdoctoral Fondecyt Research Project 3120141.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4037. doi:1538-7445.AM2012-4037
Abstract 4042: Loss of imprinting at the 14q32 locus contributes to osteosarcoma
Authors: Shu Jingmin , Li Lihua , Moriarity Branden , Thayanithy Venugopal , Spector Logan G. , Largaespada David A. , Steer Clifford J. , Subramanian Subbaya 
Published: August 2012

Abstract
Osteosarcoma (OS) is the most common primary bone malignancy affecting children and young adolescents. Downregulation of microRNAs at 14q32 locus is implicated in the pathogenesis of OS. However, array comparative genome hybridization did not show any DNA copy number loss at this locus. We hypothesized that epigenetic alterations may regulate genes and microRNAs present in this locus and contribute to the development and progression of OS. Here, we show that OS is characterized by a selective loss of imprinting (LOI) at 14q32 locus. LOI at the 14q32 locus was unique to OS, compared with 10 other cancer cell lines representing 6 tissue types. Furthermore, the average demethylation at 14q32 locus was 24%, but the global demethylation as determined by LINE-1 retrotransposons was only 7%. We also found LOI in H19, MEST, and PEG3 loci in OS samples, albeit less frequently. LOI at the 14q32 locus is highly prevalent (82.6%, 19 out of 23) only in OS patients with early-onset (12-30 years of age) of the disease. Imprinted genes at 14q32 locus were downregulated in OS due to histone modifications and the locus was controlled by bivalent domain enriched with both K4-3m and K27-3m. We determined the tumor suppressor function of this locus by overexpression of 3 imprinting genes, DLK1, RTL1 or DIO3 in SaoS2 cells. We found that all 3 genes inhibited cell proliferation and enhanced apoptosis. We also tested methylation patterns using matched buccal DNA samples obtained from OS patients and their parents. Interestingly, loss of imprinting at 14q32 locus was found in most of the OS patients and their parents, suggesting that children might be predisposed to developing OS by inheriting the instability of imprinting at this genomic site. To determine if loss of imprinting at 14q32 locus is the driving mechanism, or a secondary genetic event, we evaluated tumor tissues obtained from p53LSLR270H/;Osx-Cre mice OS models. These mice developed OS at 8-12 months (potentially representing the late-onset (&gt; 30yrs) of human OS). LOI was found only in 33% of mice OS samples, which is in agreement with the low rate of LOI detected in late-onset human OS. In conclusion, our findings suggest that LOI at the 14q32 genomic locus is likely a contributing event in the pathogenesis of OS.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4042. doi:1538-7445.AM2012-4042
Abstract 4059: Development of a point-of-care device for circulating tumor cell/cluster detection
Authors: Zhou Yang , Harb Wael A. , Shao Guoqiang , Hong Bin 
Published: August 2012

Abstract
Circulating tumor cell (CTC) has recently grabbed numerous headlines with the promise for personalized cancer diagnostics and treatment. Its clinical value, in terms of presence and the number, has been demonstrated in accumulated results for prognosis of cancer progression and mortality, and prediction of therapy response. CTCs, the leukemic phase of solid tumor, further give us the hope for a rapid and affordable optical imaging tool as CTCs are readily accessible and optically observable. Fluorescence based cell imaging has been widely spread to detect CTCs. However, it is still a challenge to accomplish a point-of-care test (POCT) due to many limitations such as the cost, fluorescence stability, variations in fluorescence detection and image processing, and data interpretation. Chromogenic labeling may offer exciting perspectives in POCT for CTC detection along with the application of metallic signal enhancement. Based on this concept, TeloVISION LLC developed a palm-sized, manually operated, POCT device for CTC detection. The device, which bears a membrane with rod-shaped, micron-sized pores, processes the whole blood without vigorous manipulation. By manually rotating the device counter-clockwise, the entire CTC assay is split into 7 steps, performed inside the device, and completed in 40 minutes. To validate the device, whole blood was withdrawn from the animals bearing metastatic tumors. Due to the restriction of the membrane, CTCs larger than 5µm size were retained while majority of the normal cells were removed due to the discrepancy in deformability. In addition, the filtration was driven by gravity so that the mechanical damage from external force was avoided. After filtration, enriched CTCs were identified by DAB stain in the presence of horseradish peroxidase (HRP) tagged anti-cytokeratin (CK) and the signal was intensified 50-fold by metal precipitation. CTCs were found in blood as well as the circulating tumor microemboli (CTM) by both chromogenic labeling and fluorescence staining (control). Heterogeneity in the CK expression was seen for both CTCs and CTM. The recovery of the device was 94% compared to that by hemocytometer. We conclude that chromogenic labeling with the signal enhancement by in situ metal precipitation can provide high-resolution, sensitive CTC detection, and prevent the false positive results as seen in fluorescent labeling due to autofluorescence and fluorescence ghost in the blood. This new portable device may provide an efficient low-cost CTC detection in clinical settings. Adjustment of the antibodies will expand the field of application such as sarcoma and melanoma, non-epithelial derived CTCs.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4059. doi:1538-7445.AM2012-4059
Abstract 4061: Computer-assisted Gleason grading of prostate cancer: Two novel approaches using nuclear shape and texture feature to classify pathologic Gleason grade patterns 3 and 4
Authors: Veltri Robert W. , Christudass Christhunesa , Epstein Jonathan I. , Ali Sahirzeeshan , Yoon Hong-Jun , Li Ching-Chung , Madabhushi Anant 
Published: August 2012

Abstract
Introduction: We used two novel computer-assisted imaging applications that incorporate nuclear shape and texture analysis to differentiate Gleason grade patterns 3 and 4 of H&Ein a tissue microarray (TMA). The two approaches utilized a variational adaptive segmentation scheme (AdACM) (Rutgers) and the cardinal multiridgelet transform (CMRT) based texture analysis (University of Pittsburgh). Methods: The TMA include 0.6 mm cores from blocks that included Gleason patterns of CaP radical prostatectomy cases from 40 patients. The TMA comprised 13 Gleason patterns 6 (3+3), 8 pattern 7 (3+4), 7 pattern 8 (4+4) and 5 pattern 9 (4+5) Gleason grade (GG) patterns. The AdACM uses 200 × 200 pixel images extracted for nuclear & architectural textural features. All features were based on mathematical energy terms with a shape prior in a multi-level set formulation. The CMRT is an image transform that is translation- and rotation-invariant, and provides a means of displaying texture contents for selection of discriminative features. With a 3-scale decomposition, it gives 8 ridgelet packets and 16 statistical texture features in a 256 x 256 pixel image patch that are averaged over 9 non-overlapping patches in a given image for separation of GG patterns 3 and 4. Results: The CMRT study utilized 3 of 16 texture features extracted from images and trained a Gaussian kernel support vector machine (SVM), a leave-one-out cross-validation showed an accuracy of 94% and AUC of 0.97. For AdACM, nuclear features and architectural features were extracted. Linear Discriminant Analysis (LDA) was employed for feature selection and best features from all three classes could discriminate Gleason grade 3 vs. 4 with an accuracy of 86% via a SVM classifier. Conclusions: Unique quantitative imaging of tissue nuclear shape and texture features can accurately differentiate Gleason patterns 3 and 4. Our research will next evaluate CaP outcomes.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4061. doi:1538-7445.AM2012-4061