bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5346: MiR-331-5p targets receptor EphA2 and attenuates tumor growth and migration in NSCLC.
Authors: Sukka-Ganesh Bhagyalaxmi , Mohammed Kamal A. , Kaye Frederic , Nasreen Najmunnisa 
Published: August 2014

Abstract
Introduction: Receptor EphA2 is over expressed in NSCLC and is associated with tumor progression and poor survival. MicroRNAs (miRs) are short, non-coding RNAs that post-transcriptionally regulate gene expression. Dysregulated expression of miRs has been observed in variety of malignancies. However, the role of miR-331-5p in NSCLC growth and migration is not known. We hypothesized that EphrinA1 treatment promotes miR-331-5p expression in NSCLC and miR-331 may inhibit tumor growth and invasion by targeting receptor EphA2.
Methods: The A-549 and H-23 NSCLC cells were treated with Ephrin-A1 for 24 hr and the miR-331-5p expression was measured by real-time PCR analysis. EphA2 protein expression was studied through Western blotting analysis. The cell proliferation was studied by WST-1 assay. The tumor formation and invasion was determined by 3-D matrigel and Wound healing assays respectively. Luciferase assay was performed to confirm miR-331-5p as one of the direct and specific target for EphA2.
Results: The miR-331-5p expression was inversely correlated with EphA2 expression. Transfection of NSCLC with precursor miR-331-5p inhibited proliferation, and migration of NSCLC cells. In addition the ability of tumor colony formation was significantly inhibited when the cells were transfected with miR-331-5p precursors. We have identified predicted target binding site of EphA2 in its 3’ UTR region by miR-331-5p. This was confirmed by Luciferase assay. Over-expression of miR-331-5p in NSCLC cells repressed the endogenous levels of EphA2 protein.
Conclusion: This study for the first time reports, that miR-331-5p directly regulates EphA2 mRNA and protein expression in NSCLC. MiR-331 directly binds to 3’ -UTR region of EphA2 and inhibits tumor growth and migration. Our findings also provide an insight into the regulation of EphA2 expression and suggest that miR-331-5p has the capacity to regulate signalling pathways critical for the development and progression of NSCLC.
Funding: RC1 from Florida Department Health, and VA Merit
Citation Format: Bhagyalaxmi Sukka-Ganesh, Kamal A. Mohammed, Frederic Kaye, Najmunnisa Nasreen. MiR-331-5p targets receptor EphA2 and attenuates tumor growth and migration in NSCLC. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5346. doi:10.1158/1538-7445.AM2013-5346
Abstract 5348: Characterization of the EZH2-MMSET histone methyltransferase regulatory axis in cancer.
Authors: Asangani Irfan A. , Ateeq Bushra , Cao Qi , Dodson Lois , Pandhi Mithil , Kunju Lakshmi P. , Mehra Rohit , Lonigro Robert J. , Siddiqui Javed , Palanisamy Nallasivam , Wu Yi-Mi , Cao Xuhong , Kim Jung H. , Iyer Mathew K. , Maher Christopher A. , Kumar-Sinha Chandan , Varambally Sooryanarayana , Chinnaiyan Arul M. , Zhao Meng , Qin Zhaohui S. 
Published: August 2014

Abstract
Histone methyltransferases (HMTases), as chromatin modifiers, regulate the transcriptomic landscape in normal development as well in diseases such as cancer. Here, we molecularly order two HMTases, EZH2 and MMSET that have established genetic links to oncogenesis. EZH2, which mediates histone H3K27 trimethylation and is associated with gene silencing, was shown to be coordinately expressed and function upstream of MMSET, which mediates H3K36 dimethylation and is associated with active transcription. EZH2 transcript expression levels were compared to all HMTases (n=51) in our compendium of RNA-seq data comprising 474 malignant and benign samples, representing 14 different tissue types. MMSET displayed the strongest correlation with EZH2 expression. Similarly, a meta-analysis of 1755 samples from 22 published microarray gene expression studies in Oncomine revealed co-expression of MMSET and EZH2 in 15 different cancers. We found that the EZH2-MMSET HMTase axis is coordinated by a microRNA network and that the oncogenic functions of EZH2 require MMSET activity. Together, these results suggest that the EZH2-MMSET HMTase axis coordinately functions as a master regulator of transcriptional repression, activation, and oncogenesis and may represent an attractive therapeutic target in cancer.
Citation Format: Irfan A. Asangani, Bushra Ateeq, Qi Cao, Lois Dodson, Mithil Pandhi, Lakshmi P. Kunju, Rohit Mehra, Robert J. Lonigro, Javed Siddiqui, Nallasivam Palanisamy, Yi-Mi Wu, Xuhong Cao, Jung H. Kim, Mathew K. Iyer, Christopher A. Maher, Chandan Kumar-Sinha, Sooryanarayana Varambally, Arul M. Chinnaiyan, Meng Zhao, Zhaohui S. Qin. Characterization of the EZH2-MMSET histone methyltransferase regulatory axis in cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5348. doi:10.1158/1538-7445.AM2013-5348
Abstract 535: IGF-I stimulates amino acid transporter xC- function to reduce intracellular ROS level and promote proliferation in human breast cancer cells.
Authors: Yang Yuzhe , Becker Marc A. , Yee Douglas 
Published: August 2014

Abstract
Our laboratory identified SLC7A11/xCT as an IGF-I-induced gene specifically regulated through the adaptor protein IRS-1 in breast cancer cells. xCT encodes the functional subunit of the heterodimeric plasma membrane transport system xC-, which is critical for the cellular uptake of cystine in exchange for intracellular glutamate. xC- transporter is involved in the regulation of proliferation, metastasis, and treatment resistance in cancer. To determine the role for xC- expression in IGF-mediated breast cancer cell biology, two classes of breast cancer cell lines were studied: estrogen receptor (ER) positive cell lines (IRS-1 activated) MCF-7, ZR-75-1, and T47D; basal-like cell lines (IRS-2 activated) MDA-MB-231, BT549, and HS578T. Significantly increased xCT mRNA expression was observed only in ER positive cell lines, which was eliminated by IRS-1 specific siRNA (25 nM). IGF-I increased xCT protein expression measured by immunblots and flow cytometry only in ER positive cells in an IRS-1 dependent manner. To measure the function of upregulated xC-, extracellular levels of glutamic acid and intracellular level of total glutathione were measured in MCF-7 and MDA-MB-231. IGF-I regulated xC- transporter function only in IRS-1 activated MCF-7 cells. In MCF-7 cells, IGF-I-stimulated monolayer and anchorage-independent growth were suppressed by treating cells with xCT shRNA or the xCT chemical inhibitor sulfasalazine (SASP, 0.1 mM). Direct measurement of reactive oxygen species (ROS) and levels of phospho-p38MAPkinase showed that IGF-I reduced cellular ROS level (induced by 10 Gy of irradiation or 1 ug/ml of mitomycin C), SASP (0.1 mM) reversed this IGF-I effect. Anchorage-independent growth assays suggested that inhibiting xC- function by 0.1 mM SASP sensitized the response of MCF-7 cells to anti-IGF-IR monoclonal antibody humanized EM164 (ImmunoGen, Inc.) and tyrosine kinase inhibitor NVP-AEW-541. These data suggest that IGF-I promotes the proliferation of ER positive breast cancer cells by regulating xC- transporter function in an IRS-1 dependent manner. Furthermore, IGF-I protects cells from ROS via upregulation of xC-. Our findings also imply that combination of xC- transporter inhibitor with anti-IGF-IR agents may have synergic therapeutic effects.
Citation Format: Yuzhe Yang, Marc A. Becker, Douglas Yee. IGF-I stimulates amino acid transporter xC- function to reduce intracellular ROS level and promote proliferation in human breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 535. doi:10.1158/1538-7445.AM2013-535
Abstract 5350: Enhancer of zeste homolog 2 (EZH2) promotes progression of cholangiocarcinoma by regulating cell cycle and apoptosis.
Authors: Nakagawa Shigeki , Okabe Hirohisa , Sakamoto Yasuo , Hayashi Hiromitsu , Hashimoto Daisuke , Kuroki Hideyuki , Imai Katsunori , Nitta Hidetoshi , Chikamoto Akira , Watanabe Masayuki , Beppu Toru , Baba Hideo 
Published: August 2014

Abstract
Background: Enhancer of zeste homolog 2 (EZH2) is a catalytic subunit of the polycomb repressive complex 2 (PRC2). When present in PRC2, EZH2 catalyzes trimethylation on lysine 27 residue of histone H3(H3K27Me3), resulting in epigenetic silencing of gene expression and cancer progression. Here, we investigated the expression and the function of EZH2 in intrahepatic and extrahepatic cholangiocarcinoma (ICC and ECC).
Methods: The influence of EZH2 on cell growth and apoptosis was assessed by knockdown experiments using siRNA in cholangiocarcinoma cell lines, RBE and TFK-1. Target genes of EZH2 in cholangiocarcinoma cell lines were searched by real time PCR. The clinical significance of EZH2 in 84 cholangiocarcinoma patients (ICC and ECC in 45 and 41, respectively) who underwent a curative surgery was examined by immunohistochemistry.
Results: In vitro analysis, a knockdown of EZH2 reduced cell growth and induced G1 arrest, and induced apoptosis confirmed by Annexin V staining and increasing sub-G1 population of two cholangiocarcinoma cell lines. Moreover, a knockdown of EZH2 increased the expression of p16INK4A and p27KIP1, in real time PCR. In immunohistochemical study, The upregulation of EZH2 was correlated with tumor diameter (P = 0.0103) in ICC, lymph node metastasis (P=0.0292) in ECC, and Ki67 index in both ICC (P=0.0364) and ECC (P=0.0017). In addition, EZH2 expression was correlated with the poor prognosis in both ICC (P=0.0447) and ECC (P=0.0227).
Conclusion: The current study demonstrates that the high expression of EZH2 results in acceleration of cell cycle and anti-apoptosis in vitro analysis, and is related to poor prognosis in patients with ICC or ECC by IHC study. These results suggest that EZH2 is a potential target for cholangiocarcinoma therapeutics.
Citation Format: Shigeki Nakagawa, Hirohisa Okabe, Yasuo Sakamoto, Hiromitsu Hayashi, Daisuke Hashimoto, Hideyuki Kuroki, Katsunori Imai, Hidetoshi Nitta, Akira Chikamoto, Masayuki Watanabe, Toru Beppu, Hideo Baba. Enhancer of zeste homolog 2 (EZH2) promotes progression of cholangiocarcinoma by regulating cell cycle and apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5350. doi:10.1158/1538-7445.AM2013-5350
Abstract 5351: Phosphorylation of EZH2 at Thr487 is a marker for good prognosis in pancreatic cancer patients.
Authors: Wei Yongkun , Chang Shih-Shin , Wang Huamin , Xia Weiya , Hung Mien-Chie 
Published: August 2014

Abstract
EZH2 is the catalytic subunit of the polycomb repressive complex 2 (PRC2) and is involved in repressing gene expression through methylation of histone H3 on lysine 27 (H3K27). EZH2 is overexpressed in multiple tumor types and correlates with poor prognosis. We have previous shown that phosphorylation of EZH2 at Thr 487 by CDK1 inhibits its methyltransferase activity. However, the status of phosphorylation of EZH2 at Thr 487 (P-EZH2-T487) and its clinical implication in cancer patients have not been reported. We thus examined P-EZH2-T487 by immunohistochemistry and its association with prognosis in pancreatic cancer patients. We found that P-EZH2-T487 can be seen in 61% (47/77) of pancreatic cancer. In univariate survival analysis, patients with high P-EZH2-T487 had significantly longer overall survival time when compared with those with low P-EZH2-T487 (P=0.032). In multivariate analysis, P-EZH2-T487 was an independent prognostic marker for overall survival in pancreatic cancer (HR=0.358, P=0.001). These results suggest that P-EZH2-T487 is a prognostic indicator for clinical outcome in pancreatic cancer patients.
Citation Format: Yongkun Wei, Shih-Shin Chang, Huamin Wang, Weiya Xia, Mien-Chie Hung. Phosphorylation of EZH2 at Thr487 is a marker for good prognosis in pancreatic cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5351. doi:10.1158/1538-7445.AM2013-5351
Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 5352: Dysregulation of the repressive H3K27 trimethylation mark in head and neck squamous cell carcinoma contributes to dysregulated squamous differentiation.
Authors: Gannon Orla M. , Merida de Long Lilia , Endo Munoz Liliana , Hazar Rethinam Mehlika , Saunders Nicholas 
Published: August 2014

Abstract
Head and neck squamous cell carcinoma (HNSCC) is one of the most prevalent cancers diagnosed worldwide and is associated with a 5-year survival rate of 55%. EZH2, a component of the polycomb repressor complex 2, trimethylates H3K27 (H3K27me3); which has been shown to drive squamous differentiation in normal keratinocytes. This study determined whether inhibition of EZH2-mediated epigenetic silencing could induce differentiation or provide therapeutic benefit in HNSCC. We determined the effects of inhibiting EZH2, by either RNA interference or pharmacologically, on HNSCC growth, viability and differentiation in vitro. Xenografts of HNSCC cell lines were used to assess efficacy of 3-Deazaneplanocin A (DZNep), an inhibitor of H3K27 trimethylation, in vivo.
EZH2 was highly expressed in HNSCC cell lines in vitro and tissue microarray analysis revealed high expression in (n= 59) in situ relative to normal oral epithelium (n=12). Inhibition of EZH2 with siRNA could induce expression of differentiation genes in differentiation-refractory squamous cell carcinoma cell lines. Differentiation-refractory HNSCC cell lines displayed persistent H3K27me3 on the promoters of differentiation genes. DZNep caused cancer-cell specific apoptosis in addition to a profound reduction in colony forming efficiency and induction of some squamous differentiation genes. Furthermore, in vivo, DZNep attenuated tumour growth in two different xenograft models, caused intra-tumour inhibition of EZH2 and induction of differentiation genes in situ.
Collectively, these data suggest that aberrant differentiation in HNSCC may be attributed to epigenetic dysregulation and suggests that inhibition of PRC2-mediated gene repression may represent a potential therapeutic target.
Citation Format: Orla M. Gannon, Lilia Merida de Long, Liliana Endo Munoz, Mehlika Hazar Rethinam, Nicholas Saunders. Dysregulation of the repressive H3K27 trimethylation mark in head and neck squamous cell carcinoma contributes to dysregulated squamous differentiation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5352. doi:10.1158/1538-7445.AM2013-5352
Abstract 5366: Chronic, low-dose treatment with sulforaphane inhibits growth of breast cells.
Authors: Silvia Ashley L. , Browne Eva P. , Arcaro Kathleen F. 
Published: August 2014

Abstract
Epidemiological data indicate that dietary consumption of cruciferous vegetables may reduce the risk of many cancers including breast cancer. Cruciferous vegetables contain high levels of glucosinolates; stable phytochemicals that when metabolized produce cancer-preventing active isothiocyanates. Isothiocyanates, such as sulforaphane (SFN) found in broccoli sprouts, are thought to prevent tumorigenesis through epigenetic mechanisms controlling the expression of tumor-related genes. Ingestion of broccoli sprouts results in rapid distribution of SFN to breast epithelium, however, the effect of chronic exposure to low levels of SFN on the growth of breast cells has not been fully explored.
In the present study we assessed the effect of SFN on the growth of an immortalized breast cell line (MCF10A), and two breast cancer cell lines; the estrogen receptor positive MCF-7 cell line, and the estrogen receptor negative TMX2-28 cell line. We first determined the seeding densities for each cell line that resulted in 80% confluence after three days of growth in T-25 flasks: MCF10A (1 x 106), MCF-7 (8 x 105), and TMX2-28 (5 x 105) per 5 mL. Next, cells from each line were plated at the predetermined seeding densities in T-25 flasks and treated with either SFN (5 μM) or vehicle (0.02% DMSO) for up to 40 days. Every third day, cell cultures were photographed, after which the cells were resuspended in culture medium, counted using a hemocytometer, and the predetermined numbers of cells (seeding numbers) were returned to T-25 flasks and treated with SFN or vehicle. Cells not needed for seeding the flasks were archived for future analysis of gene expression and DNA methylation.
Treatment with SFN decreased cell growth. After 22 days of treatment with SFN, mean proliferation of both the MCF10A and MCF-7 cell cultures was inhibited by 62% (t = 2.8; p = 0.02) and 72% (t = 2.6; p = 0.02), respectively. In contrast, the decrease in proliferation of TMX2-28 cells treated with SFN was not statistically significant (t = 1.2; p = 0.255) from the vehicle control at 22 days. However, by day 40, TMX2-28 cell cultures treated with SFN showed a significant decrease in mean proliferation 36% (t = 2.2; p = 0.04). Ongoing studies are examining the epigenetic mechanisms by which SFN decreased cell proliferation.
Citation Format: Ashley L. Silvia, Eva P. Browne, Kathleen F. Arcaro. Chronic, low-dose treatment with sulforaphane inhibits growth of breast cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5366. doi:10.1158/1538-7445.AM2013-5366
Abstract 5376: Expression of histone deacetylases (HDACs) 1, 2, 3 and 7 in pancreatic cancer cell lines: The role of indole-3-carbinol.
Authors: Lyn-Cook Beverly D. , Pang Li , Lyn-Cook Lascelles E. , Word Beverly , Hammons George 
Published: August 2014

Abstract
Pancreatic cancer has the poorest prognosis of most cancers, mainly due to its resistant phenotype. Early or late biomarkers are greatly needed in this cancer. Scientists have shown that epigenetic mechanisms may be involved in the etiology and progression of pancreatic cancer. Epigenetic changes involve modification of the DNA through hyper- and hypo-methylation, histone modification and modulation in expression of microRNAs. Histone deacetylases modify tails on the histones, which can result in repressive chromatin formation and inactivation of specific genes. This study, using real-time PCR, examined the level of HDACs 1, 2, 3 and 7 in four pancreatic cancer cell lines (BXPC, CFPAC,ASPC and MIA). The highest level of expression of HDAC 1 and 3 was found in CFPAC and MIA cells compared to BXPC and ASPC cells. Indole-3 carbinol, in previously published studies from our laboratory, have shown several anti-cancer activities, including the re-expression of p16 in MIA cells (Lyn-Cook et al, 2010 Anticancer Res., 30:4907-4914). The current study demonstrated that indole-3-carbinol at concentrations of 20 μM and 100 μM significantly decreased expression of HDAC 3 by 2-fold and 3-fold in MIA cells, respectively. Results further showed that indole-3 carbinol decreased expression of HDAC 1 in CFPAC cells and HDAC 7 in BXPC cells by 2-fold. No change in expression was shown in any of the cell lines on HDAC 2. This study showed that indole-3-carbinol may exert histone deacetylase inhibitory effects. Further studies are needed to confirm these results. Specific HDACs as potential targets for pancreatic cancer are growing among the scientific community. HDACs are known to be regulators of growth, differentiation and cell death and dysfunction of transcriptional repression mediated by HDACs may lead to the development of cancer.
Citation Format: Beverly D. Lyn-Cook, Li Pang, Lascelles E. Lyn-Cook, Beverly Word, George Hammons. Expression of histone deacetylases (HDACs) 1, 2, 3 and 7 in pancreatic cancer cell lines: The role of indole-3-carbinol. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5376. doi:10.1158/1538-7445.AM2013-5376
Abstract 5377: Profiling of aurora kinase signaling in microdissected gastric cancer samples indicates a significant increase in histone deacetylase 4 (HDAC4).
Authors: Colarossi Lorenzo , Memeo Lorenzo , Liotta Lance A. , Espina Virginia , Mueller Claudius 
Published: August 2014

Abstract
Introduction & Methods: Gastric cancer represents one of the leading causes of cancer related death in the world second only to lung cancer resulting in 800,000 deaths per year. Aurora kinases, a family of serine/threonine kinases essential for cytokinesis and chromosome segregation, are frequently over-expressed in various cancer types. Moreover, Aurora kinase has recently been shown to directly regulate epigenetic changes (Guise et al., Mol Cell Proteomics. 2012 Nov;11(11):1220-9). Aurora kinase signaling may also impact E-cadherin function via a GSK3-α/β induced increase in β-catenin. E-cadherin deregulation has been implicated in gastric cancer, especially in diffuse types where it is mutated in many cases.
We have profiled Aurora kinase signaling pathway activation and its effects on epigenetic and cytoskeletal changes in gastric cancer. Human gastric cancer tumor samples (n=10; 4 diffuse, 5 intestinal, 1 undifferentiated adenocardinoma) and patient matched normal mucosa were procured by laser capture microdissection. Reverse phase protein microarrays were used to quantify total and post-translationally modified proteins in cell signaling pathways directly and indirectly related to Aurora kinase.
Results: We found a significant increase in histone deacetylase 4 levels (HDAC4, p&lt;0.0001) concomitant with increased phosphorylation of Aurora kinase (p&lt;0.1) and a decrease in phosphorylated GSK3-α/β (p&lt;0.05) in gastric tumor epithelia compared to patient matched normal mucosa. Gastric cancer samples of diffuse type showed a significantly reduced expression of E-Cadherin (p&lt;0.05) as compared to intestinal type gastric cancer. In contrast, patient samples with lymph node metastasis (n=6) were associated with an increased level of phospho-GSK3-α/β in both normal mucosa (p&lt;0.05) and matched tumor epithelia (p&lt;0.1).
Conclusions: To our knowledge, this is the first time that increased levels of HDAC4 have been reported in gastric cancer. The demonstrated increase in Aurora kinase phosphorylation in gastric tumor epithelia may provide a mechanistic link to the observed HDAC4 upregulation.
Citation Format: Lorenzo Colarossi, Lorenzo Memeo, Lance A. Liotta, Virginia Espina, Claudius Mueller. Profiling of aurora kinase signaling in microdissected gastric cancer samples indicates a significant increase in histone deacetylase 4 (HDAC4). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5377. doi:10.1158/1538-7445.AM2013-5377