bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3483: ESR1 gene expression and expression ratio of ESR1 to ESR2 are associated with worse prognosis in papillary thyroid carcinoma
Authors: Yi Jin Wook , Song Ra-Yeong , Yu Hyeong Won , Lee Joon-Hyop , Kwon Hyungju , Chai Young Jun , Choi June Young , Kim Su-jin , Lee Kyu-Eun 
Published: July 2016

Abstract
Introduction
Expression of estrogen receptor (ER) and progesterone receptor (PR) in breast cancer have been widely studied and used as prognostic markers or therapeutic targets in breast cancer patients. However, clinical and pathologic roles of these hormone receptor genes have not been clearly established in thyroid cancer.
Materials and methods
To evaluate the relationship between sex hormone receptor gene expressions and clinicopathologic variables, we analyzed ESR1, ESR2, ESR ratio (ESR1/ESR2) and PGR mRNA expression data from The Cancer Genome Atlas (TCGA) RNA sequencing experiment in papillary thyroid carcinoma (PTC). To validate results from TCGA analysis, we additionally analyzed a public microarray data (GSE 54598) from the gene expression omnibus (GEO) database.
Results
In TCGA data analysis, ESR1, ESR ratio and PGR expression were significantly higher in PTC tissue than in normal thyroid tissue (Mean value of normal versus tumor were 264.045 vs 659.427 in ESR1, 19.064 vs 92,017 in ESR ratio and 58.514 vs 119.792 in PGR, respectively). In female patients, ESR1 and ESR ratio was negatively correlated with advanced age (t-value was -2.257 and -2.949 in linear regression analysis). Expression of ESR1 and ESR ratio were higher in classic type PTC, presentation of lymphovascular invasion and BRAF V600E mutation. Higher expression group of ESR1 and ESR ratio showed worse overall survival in female PTC patients (Hazard ratio was 6.348 in ESR1 and 4.031 in ESR ratio, respectively). In microarray analysis, ESR1 and ESR ratio expressions were higher in tumor than normal, classic than follicular variant and BRAF V600E than wild type.
Conclusion
Expression of ESR1 and ESR ratio were associated with aggressive prognostic factors and worse overall survival in female PTC patients. These results suggest that higher expression of ESR1 and ESR ratio can be used as prognostic markers to predict survival in female patients.
Citation Format: Jin Wook Yi, Ra-Yeong Song, Hyeong Won Yu, Joon-Hyop Lee, Hyungju Kwon, Young Jun Chai, June Young Choi, Su-jin Kim, Kyu-Eun Lee. ESR1 gene expression and expression ratio of ESR1 to ESR2 are associated with worse prognosis in papillary thyroid carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3483.
Abstract 3501: Puma/pBcl-2 axis influences sensitivity of cancer cells to multiple stimuli
Authors: Liu Yubo 
Published: July 2016

Abstract
Bcl-2 is a central regulator of cell survival that is overexpressed in the majority of cancers cells.Previous reports indicated that Bcl-2 phosphorylation (S70) may provide a switch by which cellscan regulate the pro-survival functions of Bcl-2. But there is still controversy over the function of phosphorylatedBcl-2 (pBcl-2).In the presentstudy, Puma/pBcl-2 axis was found to play a key role in regulating the sensitivity of cancer cells to multiple stimuli.Based on immunoprecipitation, weobserved that the pro-apoptoticBH3-only protein Puma which was induced by paclitaxel competitively bound pBcl-2 (S70) and displaced Bim and Bax from pBcl-2(S70) in Hela and HL60 cells.Further study showed thatdepletion of Puma (siRNA), but not other BH3 only proteins, significantlydecreased paclitaxel-induced apoptosis.Bim/pBcl-2 and Bax/pBcl-2 complexeswere largely maintained when Pumawas reduced. These data indicated that the phosphorylation of Bcl-2(S70) enhancedits anti-apoptotic function and the induction of Puma could neutralize this effect. Similarly,the induced Puma was also responsible for Bim and Bax displacement from pBcl-2 (S70) when Hela and HL60 cellswere treated with DNA damage agents(cisplatin and etoposide).However, Pumacannot displace Bax or Bim when pBcl-2 (S70) was redundant in the cells expressedphospho-mimetic S70E-Bcl-2. It demonstrated that DNA damage induced apoptosis could be inhibit by pBcl-2 (S70) and the induction of Puma could antagonize pBcl-2 (S70). In a cell free system, mitochondria from Hela and HL60 cells which expressing the S70E-Bcl-2 wereincubated with different BH3 peptides. Cytochrome c release couldbe induced by a low concentration of Puma-BH3 peptide, but a higher concentrationwas necessary during Bimengagement.By contrast, Bad-BH3 and Noxa-BH3cannot initiate the release of cytochrome c.In conclusion, multiple stimuli activates the mitochondrial apoptoticpathway in cancer cells, in which Puma/pBcl-2(S70) axisplay a key role in a novel mechanism involving the displacement of Bim and Bax from pBcl-2 (S70)by Puma. In contrast with the debatable function of Bcl-2 phosphorylation (S70), this study provided evidence that the phosphorylation of Bcl-2 at S70 enhanced cell survivalin multiplestimuli triggered apoptosis.
Citation Format: Yubo Liu. Puma/pBcl-2 axis influences sensitivity of cancer cells to multiple stimuli. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3501.
Abstract 3535: Induction of MAPK- and ROS-dependent autophagic cell death in gastric carcinoma by combination of bortezomib and romidepsin
Authors: Chiang Alan K. , Yeung Pauline L. , Hui Kwai Fung 
Published: July 2016

Abstract
Proteasome inhibitors and histone deacetylase (HDAC) inhibitors can synergistically induce apoptotic cell death in certain cancer cell types but their combinatorial effect on the induction of autophagy remains unknown. Here, we investigated the combinatorial effects of a proteasome inhibitor, bortezomib, and an HDAC inhibitor, romidepsin, on the induction of apoptotic and autophagic cell death in gastric carcinoma (GC) cells. Isobologram analysis showed that low nanomolar concentrations of bortezomib/romidepsin could synergistically induce killing of GC cells. The synergistic killing was due to the summative effect of caspase-dependent intrinsic apoptosis and caspase-independent autophagy. The autophagic cell death was dependent on the activation of MAPK family members (ERK1/2 and JNK), and generation of reactive oxygen species (ROS), but was independent of Epstein-Barr virus infection. In vivo, bortezomib/romidepsin also significantly induced apoptosis and autophagy in GC xenografts in nude mice. This is the first report demonstrating the potent effect of combination of HDAC and proteasome inhibitors on the induction of MAPK- and ROS-dependent autophagy in addition to caspase-dependent apoptosis in a cancer type.
This project is funded by CRCG (#104003676) grant of KFH, CRCG (#104002845) and Epstein-Barr virus research (# 200004525) grants of AKSC.
Citation Format: Alan K. Chiang, Pauline L. Yeung, Kwai Fung Hui. Induction of MAPK- and ROS-dependent autophagic cell death in gastric carcinoma by combination of bortezomib and romidepsin. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3535.
Abstract 3543: Alternating electric fields (TTFields) induce autophagy in human cancer cell lines
Authors: Porat Yaara , Shteingauz Anna , Giladi Moshe , Schneiderman Rosa S. , Voloshin Tali , Munster Mijal , Blat Roni , Kirson Eilon D. , Weinberg Uri , Palti Yoram 
Published: July 2016

Abstract
Tumor treating fields (TTFields) are an established anti-neoplastic treatment modality in patients with glioblastoma. TTFields are delivered via noninvasive application of low-intensity, intermediate-frequency, alternating electric fields to the region of the tumor. Previous studies have shown that TTFields lead to increased granularity in treated cells. Granular or vesicular structures can indicate the formation of autophagosomes which are key structures in autophagy. Autophagy has been shown to regulate cell survival and proliferation under stress conditions and, in certain cases, influence cellular response to cytotoxic drugs. The goal of this study was to evaluate the possible effect of TTFields on the induction of autophapgy in treated cells.
Different cancerous cell lines were treated with TTFields using the inovitro system. Cellular granularity was evaluated using flow cytometry. Autophagy was monitored by quantifying Lipidated Microtubule Associated Protein Light Chain 3 (LC3-II, a consensus marker for autophagosomes) levels using immunoblotting and immunofluorescence microscopy. Transmission Electron Microscopy (TEM) was used to visualize autophagosome-like structures.
Flow cytometry analysis demonstrated that TTFields application leads to a significant increase in cellular granularity in all tested cell lines (up to 40%, P&lt;0.05). Significant elevation in LC3-II levels was observed in treated U-87 MG cells using immunoblotting analysis (37%, P = 0.01). Evidence of increased autophagy following TTFields application was also detected using fluorescence microscopy, where punctate distribution of LC3-II was observed. TEM micrographs demonstrated the presence of autophagy typical, autophagosome-like structures in TTFields treated cells.
TTFields are known to exert anti-mitotic effects by disrupting highly dipolar structures which play critical roles in mitosis. Our results suggest that in addition, TTFields can also induce cellular autophagy. Future studies are warranted to examine to what extent TTFields-elicited- autophagy may affect treatment outcomes, and to investigate the therapeutic implications of combining TTFields with autophagy inhibitors.
Citation Format: Yaara Porat, Anna Shteingauz, Moshe Giladi, Rosa S. Schneiderman, Tali Voloshin, Mijal Munster, Roni Blat, Eilon D. Kirson, Uri Weinberg, Yoram Palti. Alternating electric fields (TTFields) induce autophagy in human cancer cell lines. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3543.
Abstract 3545: Gallotannin induces senescence and autophagy leading to cell death via SIRT1 inhibition and AMPK activation in hepatocellular carcinoma cells
Authors: Kwon Hee Young , Kim Sung-hoon 
Published: July 2016

Abstract
Hepatocellular carcinoma (HCC) is one of the most fatal malignancies with high mortality rate worldwide. In the present study, the underlying antitumor mechanism of gallotannin, a hydrolysable tannic acid, was elucidated in HepG2 and SKHep1 HCC cells. Gallotannin suppressed growth and colony formation and also induced senescence via upregulation of p21, G0 / G1 arrest and higher senescence-associated β-galactosidase (SA-β-gal) activity within 24 h culture in HepG2 and SKHep1 cells. Of note, gallotannin also induced autophagy by increasing the number of protein-cytosol-associated protein light chain 3 (LC3) punctae, LC3B-II conversion and SQSTM1/p62, autophagic vacuoles (AVOs) by transmission electron microscopy (TEM) and decreasing the of Beclin1 in HepG2 and SKHep1 cells. A tandem fluorescent-tagged LC3 reporter plasmid (GFP-mRFP-LC3) transfection revealed that gallotannin increased the number of yellow colored LC3 punctae through the co-localization of GFP and mRFP punctae and yellow colored LC3 punctae were increased by CQ or NH4Cl with accumulation of autophagosomes, LC3B-II and SQSTM1/p62 in HepG2 and SKHep1 cells. Additionally, gallotannin attenuated the of sirtuin1 (SIRT1) and mTOR and activated the phosphorylation of 5′-AMP activated protein kinase (AMPK) in two HCC cells. Furthermore, AMPK activator AICAR significantly enhanced SA-β-gal activity and antiproliferation induced by gallotannin, while AMPK inhibitor compound C did not in two HCC cells. Of note, gallotannin time dependently activated caspase 8 / 3 and time/concentration dependently increased sub G1 portion along with weak cleavages of PARP in two HCC cells. Collectively, gallotannin induces senescence and inhibits late autophagy flux, finally leading to apoptotic cell death in HepG2 and SKHep1 cells via inhibition of SIRT1 and activation of p-AMPK as a potent antitumor agent for HCC treatment.
Citation Format: Hee Young Kwon, Sung-hoon Kim. Gallotannin induces senescence and autophagy leading to cell death via SIRT1 inhibition and AMPK activation in hepatocellular carcinoma cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3545.
Abstract 3550: Activation of a novel form of regulated necrosis to eliminate extracellular matrix detached epithelial cells
Authors: Hawk Mark A. , Buchheit Cassandra L. , Shen Luqun , Fagan Patrick , Schafer Zachary T. 
Published: July 2016

Abstract
The overwhelming majority of breast cancer deaths are caused by the metastasis of cancer cells from the primary tumor to distant sites in the body. For cancer cells to successfully metastasize, they must: detach from the primary tumor, move out of the original tissue into the circulatory or the lymphatic system, travel to a new site followed by arresting their movement, and then extravasate to begin colonizing a secondary site. When normal epithelial cells detach from the extracellular matrix (ECM), they induce caspase-dependent cell death which is known as anoikis. Anoikis can therefore serve as a critical barrier to metastasis as cancer cells are exposed to limited and variable matrix conditions during each step of the metastatic cascade. However, our previous studies suggest that anoikis evasion is not sufficient to protect ECM-detached cells from cell death. Using MCF10A mammary epithelial cells, we have previously shown that ECM-detachment induced metabolic changes can compromise the survival of detached cells, although the precise mechanism controlling cell death remains unclear. Here, we present data suggesting that ECM-detached cells can also be eliminated by regulated necrosis (RN), a genetically programmed, caspase-independent form of necrosis that is morphologically indistinguishable from classical necrosis. In general, the molecular mechanisms involved in RN are poorly understood. With this in mind, the current most appreciated subtype of RN, termed necroptosis, is dependent upon TNFα, RIP1K, RIP3K, MLKL, and PGAM5 to execute cell death. Our data in ECM-detached cells suggest that cells are being eliminated by a mechanism that is dependent on the kinase activity of RIP1K. Further studies have shown an increase in the expression of CYLD, a deubiquitinating enzyme that specifically removes ubiquitin chains from RIP1K, in turn stabilizing both RIP1K expression and activation. Upon analyzing other dependent executioners of necroptosis, we have strikingly found that TNFα, RIP3K, and MLKL are all dispensable for RN to occur in ECM-detachment. Furthermore, using a variety of molecular tools, such as 3D cell culture, we have found that PGAM5 is necessary for the execution of ECM-detachment induced RN. These findings uncover a novel and distinct RN pathway and highlight the need to more thoroughly understand RN as well as the mechanisms employed by ECM-detached cells to antagonize RN and promote their survival in detachment.
Citation Format: Mark A. Hawk, Cassandra L. Buchheit, Luqun Shen, Patrick Fagan, Zachary T. Schafer. Activation of a novel form of regulated necrosis to eliminate extracellular matrix detached epithelial cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3550.
Abstract 3562: BCR/ABL increases EZH2 levels which regulates XIAP expression via miRNA-219 in chronic myeloid leukemia cells
Authors: Ikezoe Takayuki , Nishioka Chie , Yang Jing , Yokoyama Akihito 
Published: July 2016

Abstract
In this study, we showed that the levels of enhancer of zeste homolog 2 (EZH2) in bone marrow mononuclear cells (BMMNCs) isolated from individuals with chronic myeloid leukemia (CML) (n = 12) were greater than those in BMMNCs isolated from healthy volunteers (n = 6). Lentiviral transduction of the BCR/ABL gene in Ba/F3 cells increased EZH2 levels in parallel with phosphorylation of STAT5. Notably, chromatin immunoprecipitation assays showed that STAT5A bound to a promoter region of the EZH2 gene, resulting in an increase in the transcriptional activity of EZH2 in leukemia cells. Importantly, downregulation of EZH2 by short hairpin RNAs (shRNAs) inhibited the expression of the antiapoptotic protein XIAP and increased the miR-219 levels associated with a decrease in hypermethylation of miR-219-1 CpG islands. Moreover, overexpression of miR-219 decreased the levels of XIAP in CML cells. Since the 3′-untranslated region (3′-UTR) of XIAP contains miR219-5p-complementary binding site, miR-219 might modulate the expression of XIAP through binding of miR-219 on the 3′-UTR of XIAP.
Taken together, BCR/ABL positively regulates the expression of EZH2 via STAT5 signaling. EZH2 modulates epigenetic changes at DNA methylated regions encoding miR-219 and downregulates the level of miR-219, resulting in upregulation of XIAP.
Citation Format: Takayuki Ikezoe, Chie Nishioka, Jing Yang, Akihito Yokoyama. BCR/ABL increases EZH2 levels which regulates XIAP expression via miRNA-219 in chronic myeloid leukemia cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3562.
Abstract 3580: Topoisomerase II mediated DNA damage generates unique classes of genome rearrangements
Authors: Gilbertson Matthew , Patel Radhika , Nitiss Karin C. , Nitiss John L. 
Published: July 2016

Abstract
Topoisomerase 2 (Top2) is the target of active anti-cancer agents such as etoposide and doxorubicin. These drugs interfere with the Top2 catalytic cycle and lead to trapping of the enzyme as a covalent protein: DNA complex. This trapped covalent complex is a unique DNA lesion that includes DNA strand breaks and covalent protein adducts at the site of the breaks. While Top2 mediated DNA damage is the major mechanism for tumor cell killing, it is also responsible for drug-induced translocations that can lead to secondary malignancies. We have developed several novel systems for assessing Top2-mediated genome instability using yeast as a model system. We developed repair proficient yeast strains that accumulate high levels of etoposide and other Top2 targeting drugs. We selected for loss-of-function mutations in the yeast CAN1 gene that arise following etoposide treatment, and examined the induced mutations by DNA sequencing. We identified a unique spectrum of mutations generated by etoposide in yeast. One novel class of mutational events induced by etoposide is relatively large deletions (&gt;300 bp) and tandem duplications. Most of these etoposide-induced events are flanked by 4-8 nucleotide direct repeats. To further characterize the mechanism of Top2 induced genome instability, we took advantage of a novel Top2 allele isolated in our laboratory. This Top2 allele, termed top2AR, exhibits elevated levels of drug independent DNA cleavage in vitro, and inviability when combined with yeast mutants defective in DNA double strand break repair. We found a similar spectrum of large deletions and tandem duplications flanked by 4-8 nucleotide direct repeats. Interestingly, we also found that the recovery of large deletions and insertions was enhanced in yeast cells lacking Tdp1, a gene that participates in the repair of protein:DNA covalent complexes. Our results illustrate a novel type of genome rearrangement that is mediated by covalent Top2 DNA damage. It is noteworthy that the deletions and insertions flanked by short direct repeats have not been previously observed with other classes of DNA damaging agents. We suggest that the unique properties of these lesions may also contribute to oncogenic translocations induced by etoposide and other Top2 targeting drugs. We are currently using next-generation sequencing of etoposide treated yeast cells and cells carrying the top2AR allele to obtain a more complete picture of the types of alterations induced by Top2 mediated DNA damage.
Citation Format: Matthew Gilbertson, Radhika Patel, Karin C. Nitiss, John L. Nitiss. Topoisomerase II mediated DNA damage generates unique classes of genome rearrangements. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3580.
Abstract 3587: Multiplexed nuclear area and micronucleus screening identifies SKP1 as a human chromosome instability gene
Authors: Thompson Laura , Baergen Allison , Lichtensztejn Zelda , McManus Kirk 
Published: July 2016

Abstract
Chromosome instability (CIN) is defined as an increase in the rate at which whole chromosomes or large chromosomal fragments are gained or lost. It is hallmark of cancer that occurs frequently in both solid and liquid tumors. In addition, CIN is associated with highly aggressive tumors, the acquisition of multi-drug resistance, tumor recurrence and poor patient prognosis. Despite this, the majority of human CIN genes have yet to be elucidated, highlighting the need for studies aimed at identifying the defective genes that underlie CIN. In this study we utilized two complementary, image-based approaches capable of detecting CIN-associated phenotypes following RNAi-based silencing of candidate CIN genes. The first assay involves quantifying nuclear areas following silencing, where changes in mean nuclear area relative to controls act as a surrogate marker of CIN. The second approach monitors micronucleus (MN) formation where increases in the number of micronuclei are indicative of DNA damage or the mitotic defects that underlie CIN. These assays were employed in a high-content screen of 164 human candidate CIN genes in two unrelated cell lines, HT1080 and hTERT. In HT1080, the nuclear area and MN enumeration assays identified 88 and 96 putative CIN genes, respectively. In hTERT, the nuclear area and MN assays identified 112 and 19 putative CIN genes, respectively. Promising putative CIN genes such as SKP1 were identified and prioritized for subsequent validation based on the number of assays that identified the gene, and the strength of the CIN phenotype. Preliminary data collected through Western blotting, mitotic chromosome spreads and flow cytometry, provides evidence to support the validation of SKP1 as a bona fide human CIN gene. Identification and characterization of human CIN genes will provide critical insights into CIN and tumorigenesis, as well as identify potential targets that could be exploited in novel, precision medicine approaches for superior cancer treatment.
Citation Format: Laura Thompson, Allison Baergen, Zelda Lichtensztejn, Kirk McManus. Multiplexed nuclear area and micronucleus screening identifies SKP1 as a human chromosome instability gene. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3587.