bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1169: Growth regulation of bladder cancer cells by compounds that affect glucose metabolism
Authors: Lea Michael A. , desBordes Charles 
Published: August 2015

Abstract
The observation that cancer cells tend to exhibit enhanced rates of glycolysis presents a target for therapeutic regulation of cancer cell growth by molecules that regulate glucose metabolism. We have extended our earlier studies on colon cancer cells with an examination in bladder cancer cells of the action of an established inhibitor of PFKFB3, namely 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one (3PO) as a single agent and in combination with other molecules that affect glycolysis and proliferation. 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB-3) is an enzyme regulating glycolysis in many tumor cells. Cell growth was monitored by staining with sulforhodamine B. Of the 8 cell lines examined, growth was most rapid in the T24 and UM-UC-3 bladder cancer cells. The slowest growth was seen with RT4, TCCSUP and HT1197 cells while intermediate growth rates were observed with 5637, HT1376, and SW780 cells. Glucose uptake from the medium and acidification of the medium over a 72-hour incubation tended to correlate with growth rate. Thus the greatest changes were seen with the T24 and UM-UC-3 cell lines and the least change was seen with RT4 and TCCSUP cells. Growth was inhibited when glutamine was omitted from the medium but growth was blocked in the absence of glucose except for RT4 cells in which growth was inhibited more in the absence of glutamine. Treatment with 10 μM 3PO as a single agent for 72 hours caused average growth inhibition in the range of 25-50% without apparent relationship to the control growth rates of the cell lines. Additive inhibitory effects on growth were observed when cells were coincubated with 3PO and either 1 mM 2-deoxyglucose, 50 μM phenformin or 1 mM butyrate. In addition to being growth inhibitory, butyrate also caused significant induction of alkaline phosphatase activity in some cell lines. When cells were incubated with phenformin as a single agent, there was generally an increased acidification of the medium that was diminished by coincubation with 3PO. In conclusion, 3PO was an inhibitor of proliferation of human bladder cancer cells and may be a useful agent in combination with other drugs that inhibit bladder cancer cell proliferation.
Citation Format: Michael A. Lea, Charles desBordes. Growth regulation of bladder cancer cells by compounds that affect glucose metabolism. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1169. doi:10.1158/1538-7445.AM2015-1169
Abstract 1174: Exploiting glioma stem cell metabolism to abrogate radioresistance in glioblastomas
Authors: Spehalski Elizabeth I. , Saito Keita , Kramp Tamalee , Tandle Anita , Rath Barbara , Cherukuri Murali , Camphausen Kevin 
Published: August 2015

Abstract
Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor in adults and, even with aggressive treatment that includes surgical resection, radiation, and temozolomide administration, prognosis is poor due to tumor recurrence. There is evidence that within GBMs a small number of glioma stem cells (GSCs) exist, which are thought to be radiation resistant and may be capable of repopulating a tumor after treatment. Like most cancers, GBMs largely employ aerobic glycolysis to create ATP, a phenomenon known as the Warburg effect. Experiments have shown that the cellular metabolism of GSCs differs from that of differentiated cells, making them an attractive target for novel therapeutic approaches. Much work has been done to analyze the metabolic profiles of GSCs with the goal of identifying potential therapeutic targets, but little data exists linking metabolic changes to radiation resistance. The purpose of this study is to determine the impact of radiation on the cellular metabolism of GSCs and to identify changes that can be exploited for treatment in conjunction with radiation therapy.
To this end, we analyzed three human GSC lines harboring unique sets of genetic mutations that were hypothesized to differentially impact cellular metabolism. Experimental approaches include in vitro examination of metabolic rates and mitochondrial stress of the GSC lines via Seahorse extracellular flux analysis with and without radiation administration as well as analysis of tumor xenografts in mice via 13C MR spectroscopic imaging using hyperpolarized [1-13C] pyruvate as a tracer. Additionally, we are employing genomic and proteomic profiling of the GSC lines both in culture and in vivo to categorize metabolic changes following radiation treatment.
We show that the basal metabolic rates of GSC lines with distinctive genetic mutations can be differentiated using Seahorse analysis. This is confirmed in vivo by 13C MR spectroscopic imaging, thus revealing a potential non-invasive diagnostic modality. The underlying mechanism behind these metabolic differences and how this relates to each GSC's response to radiation is currently being examined. We expect that this data will result in the identification of molecular targets within the metabolic pathways of cells that can be exploited with drugs in conjunction with radiation therapy to improve the prognosis and survival of patients presenting with GBMs in the clinic.
Citation Format: Elizabeth I. Spehalski, Keita Saito, Tamalee Kramp, Anita Tandle, Barbara Rath, Murali Cherukuri, Kevin Camphausen. Exploiting glioma stem cell metabolism to abrogate radioresistance in glioblastomas. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1174. doi:10.1158/1538-7445.AM2015-1174
Abstract 1197: Selenium-binding protein 1-mediated tumor suppression is associated with alterations of lipid/glucose metabolic pathways in vivo
Authors: Ying Qi , Ansong Emmanuel , Diamond Alan M. , Lu Zhaoxin , Bie Xiaomei , Yang Wancai 
Published: August 2015

Abstract
Selenium-binding protein 1 (SBP1) is significantly decreased in most of human cancers including liver, prostate, breast, lung and colorectal cancers, and the decreased SBP1 is associated with poor clinical outcomes and recurrence, showing a tumor suppressor role. However, the underlying mechanisms are still unclear. In this study, we constructed a doxycycline inducible SBP1 plasmid (pRetroX-Tight- Pur-SBP1) and established a stable colorectal cancer cell line HCT116-TetSBP1, then injected the HCT116-TetSBP1 cell line into BALB/c CD-1 Nu female mice subcutaneously on the right flank, the control cell line HCT116-Act was injected in the left flank. The mice was supplied with Doxycycline (200 μg/mL) in drinking water. Four weeks after injection, the tumors from both sides were isolated and the weight (gm) and volume (mm3) of tumors were determined. Tumors were stored at -80°C until used for immunoblot and isobaric tags for relative and absolute quantitation (iTRAQ) analysis. We found that induction of SBP1 showed significant inhibition of cancer cell growth and metastasis in mice. iTRAQ analysis showed 132 differentially expressed proteins in response to SBP1 induction, among them, 53 proteins were upregulated and 79 proteins were downregulated. Importantly, many of the differentially altered proteins were associated with lipid/glucose metabolism. Although all seven proteins related to glucose metabolism (GAA, GAPDH, ALDH2, Thioredoxin, ENO3, UGDH and Lumican) were downregulated by SBP1 induction, thirteen lipid metabolism-related proteins act differentially on SBP1-medited signal transduction (five proteins were upregulated: DKK1, HSP60, DHCR7, ANXA4 and AGPAT5, and eight proteins were downregulated: LPCAT2, GPX5, GAPDH, NME2, ALDH2, BPIFA3, PPIA and FABP4). Bioinformatic analysis showed that these proteins were involved in Glycolysis, MAPK, Wnt, NF-kB, NOTCH and epithelial-mesenchymal transition (EMT) signaling pathways. Taken together, our results have revealed a novel mechanism that SBP1-mediated cancer inhibition is through altering lipid/glucose metabolic signaling pathways.
Note: This abstract was not presented at the meeting.
Citation Format: Qi Ying, Emmanuel Ansong, Alan M. Diamond, Zhaoxin Lu, Xiaomei Bie, Wancai Yang. Selenium-binding protein 1-mediated tumor suppression is associated with alterations of lipid/glucose metabolic pathways in vivo. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1197. doi:10.1158/1538-7445.AM2015-1197
Abstract 1205: Identification of a novel adjuvant therapeutic agent for obesity related pancreatic cancer
Authors: Harbuzariu Adriana , Garrison Robin C. , Daley-Brown Danielle S. , Beech Derrick J. , Cason Frederick D. , Harmon Tia L. , Yang Lily , Gonzalez-Perez Ruben R. 
Published: August 2015

Abstract
PURPOSE: Pancreatic cancer (PC) is an aggressive disease that develops in a symptom-free manner and is usually advanced at the time of diagnosis; the overall 5 year survival rate is less than 5%. Obesity is pandemic in the US and strongly linked to higher incidence of many cancer types. Obese people are 20% more likely to develop PC. We hypothesize that high level of leptin shown by obese PC patients could increase tumor progression and drug resistance. METHODS: A panel of PC cell lines (Panc-1, BxPC-3 and MiaPaCa-2) was challenged with leptin, a specific leptin inhibitor coupled with nanoparticles (IONP-LPrA2) and various chemotherapeutic agents (Gemcitabine, Cisplatin, Paclitaxel and Sunitinib). Cell viability, proliferation and the expression levels of molecular markers were analyzed. Further, we investigated the ability of PC cells to form tumorspheres when treated with leptin and IONP-LPrA2. The number of colonies was counted and pancreatic cancer stem cells (PCSC) markers (ALDH1, CD44, CD24, ESA) and OB-R expression was assessed. PCSC markers were determined in PC tissue arrays. RESULTS: PCSC markers were expressed in obese PC patients. Leptin induced S phase progression, overexpression of Notch and PCSC markers and increased stemness and tumorigenesis of PC cells. In addition, leptin impaired the efficacy of chemotherapeutic agents on PC cells and derived tumorspheres. The inhibition of leptin signaling via IONP-LPrA2 abrogated PC growth, reduced PCSC markers expression and enhanced chemotherapeutic action by inducing apoptosis. Our data suggests that leptin signaling is essential for PC cells proliferation and survival and reduces chemotherapeutic effects. These findings are of great importance for obese PC patients which show the highest level of leptin, poor prognosis and chemotherapy outcome. LPrA2 has no toxicity and it has been shown that it does not affect health status, appetite, body weight or energy balance in mice. IONP-LPrA2 could be a novel adjuvant for PC treatment, especially for obese PC patients. Acknowledgements: this work was supported by NIH/NCI 5SC1CA138658-05 and the DOD Breast Cancer Idea Award (W81XWH13-1-0382) to RRGP; NIH/2G12RR003034-26 and U54 MSM/UAB Cancer Center Partnership. This research was also supported by facilities and support services at Morehouse School of Medicine (NIH 1C06 RR18386).
Citation Format: Adriana Harbuzariu, Robin C. Garrison, Danielle S. Daley-Brown, Derrick J. Beech, Frederick D. Cason, Tia L. Harmon, Lily Yang, Ruben R. Gonzalez-Perez. Identification of a novel adjuvant therapeutic agent for obesity related pancreatic cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1205. doi:10.1158/1538-7445.AM2015-1205
Abstract 1207: Nrf2 suppression results in growth inhibition and enhances response to chemotherapy and radiation therapy in triple-negative breast cancer cells
Authors: Yang Yuzhe , Yee Douglas 
Published: August 2015

Abstract
Nuclear factor-erythroid 2-related factor 2 (Nrf2) is a key transcriptional activator that specifically binds to the Antioxidant Response Element (ARE) to initiate expression of various anti-oxidative and anti-inflammation genes to protect cells from oxidative stress. Recently, Nrf2 has been shown to redirect glucose and glutamine into anabolic pathways to promote cell proliferation. Constitutive stabilization of Nrf2 has been observed in many human cancers and confers chemo- and radio-resistance of cancer cells. We examined Nrf2 expression and function in triple negative breast cancer cells (TNBC). mRNA and protein expression of Nrf2 were higher in the TNBC cell lines MDA-MB-231 and MDA-MB-436 compared to immortalized breast epithelial cell lines (MCF10A and MCF12A) and estrogen receptor positive breast cancer cell lines (MCF-7 and T47D). shRNA knock-down of Nrf2 in MDA-MB-231 and MDA-MB-436 TNBC cells showed decreased mRNA expression of multiple Nrf2 regulated anti-oxidant (SLC7A11, NQO-1, GCLC, GCLM, SLC7A5, etc) and pentose phosphate pathway genes (G6PD, PGD, ME-1, TKT, TALDO1, PPAT, etc), enhanced basal levels of cellular reactive oxygen species, impaired mitochondrial function as determined by measuring mitochondrial potentials in living cells, and reduced S phase entry. Cells with decreased Nrf2 had reduced cell growth in monolayer and anchorage independent growth assays. In addition, Nrf2 suppression reduced cell migration. Nrf2 down-regulated MDA-MB-231 cells also showed increased response towards ionizing radiation and mitomycin C treatment in clonogenic and soft agar assays. Furthermore, reduced Nrf2 expression decreased the number of stem-like (CD44+/CD24-) cells in MDA-MB-231 cell populations. Thus, Nrf2 regulated many aspects of the malignant phenotype in TNBC and suggest that inhibition of Nrf2 might be a therapeutic target for TNBC.
Citation Format: Yuzhe Yang, Douglas Yee. Nrf2 suppression results in growth inhibition and enhances response to chemotherapy and radiation therapy in triple-negative breast cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1207. doi:10.1158/1538-7445.AM2015-1207
Abstract 1210: Biopterin metabolism drives tumor progression
Authors: Rabender Christopher , Alam Asim , Waters Mike , Mikkelsen Ross 
Published: August 2015

Abstract
Previous studies demonstrate that nitric oxide synthases (NOS’s) can become uncoupled during inflammatory disease states. A hallmark of this uncoupling is superoxide production from NOS in lieu of nitric oxide (NO) resulting from a reduced tetrahydrobiopterin:dihydrobiopterin ratio (BH4:BH2 ratio). As BH4 is a necessary cofactor for NO production from NOS, when this ratio is reduced, NOS becomes a peroxynitrite synthase enhancing the pro-inflammatory/pro-survival tumor environment. Our lab demonstrated that NOS is uncoupled in a variety of tumor cells both in vitro and in vivo, as well as in paired human colorectal samples due to a reduced BH4:BH2 ratio and that recoupling NOS in via exogenous BH4 both in vitro and in vivo leads to decreased tumor cell proliferation. To examine the mechanism behind the reduced BH4:BH2 ratio we used the DSS/AOM model of colitis and colon tumorigenesis. Here we demonstrate that BH4:BH2 ratio is reduced in mouse colon tumors vs mouse normal colon tissue, 1.36±0.35 vs 7.1±0.6. We determined that this reduced ratio is due at least in part to direct oxidation of BH4 to BH2, as treatment of colon cells with an NADPH Oxidase inhibitor increased the BH4:BH2 two-fold; however, this is unable to account for the degree of uncoupling associated with tumor cells. Given that inflammation appears to drive NOS uncoupling, we extracted colons from DSS/AOM treated animals and preferentially harvested colonic epithelial cells and colonic tumor cells at different stages of inflammation to assess the molecular and biochemical changes in molecules important in the regulation and production of BH4. The molecules we focused on were: GTP cyclohydrolase-1 (GCH1, rate limiting enzyme in de novo BH4 synthesis), GTP cyclohydrolase 1 feedback regulatory protein (GCHFRP, regulates GCH1 expression), quinoiddihydropteridine reductase (QDPR, necessary for salvage pathway) and sepiapterin reductase (SR, necessary for both de novo synthesis and salvage of BH4). Transcript levels of these molecules in DSS treated epithelial cells showed that GCH1 and SR were upregulated but QDPR was significantly downregulated when compared with untreated cells, potentially leading to an increase in total BH4 but once oxidized to BH2, a decrease in the salvage back to BH4. Our current research investigates the protein levels as well biochemical activity of these molecules in DSS treated epithelial cells, as well as tumor cells compared with normal cells to further understand the molecular component of the decrease in BH4:BH2 ratio in tumors.
Note: This abstract was not presented at the meeting.
Citation Format: Christopher Rabender, Asim Alam, Mike Waters, Ross Mikkelsen. Biopterin metabolism drives tumor progression. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1210. doi:10.1158/1538-7445.AM2015-1210
Abstract 1211: Metabolite assays to illuminate cellular energy networks
Authors: Sobol Mary , Vidugiriene Jolanta , Leippe Donna , Vidugiris Gediminas , Zhou Wenhui , Cali James 
Published: August 2015

Abstract
Every organism requires cellular energy metabolism to meet developmental needs. When cells change from a resting to a proliferative state, the metabolic pathways are reprogrammed to meet the demands of proliferation. Diseases such as cancer can occur when changes in the metabolic pathways support unwanted functions such as uncontrolled proliferation.
Assays for analyzing metabolites and metabolic enzymes are needed to understand cellular metabolic requirements and energy network regulation under normal and disease conditions.
We exploited the robustness, sensitivity and broad dynamic range of bioluminescence detection to develop assays for the key metabolites lactate, glucose, glutamate and glucose-6-phosphate. The technology involves using specific dehydrogenases to couple metabolite levels to NAD(P)H production. In the presence of NAD(P)H, a novel proluciferin is converted to a substrate for luciferase resulting in light signals proportional to the starting metabolite concentration. Samples (e.g. mammalian cells, culture media, tissues, 3D microtissues) are added to the metabolite detection reagent in a 1:1 ratio. An inactivation solution added to complex biological samples lowers the background and increases sensitivity by lysing the cells, inactivating cellular enzymes, and destroying endogenous NAD(P)H.
The sensitivity (1-5 pmol/sample) and broad assay window (maximum signal-to-background &gt; 100 fold) of the assays allows simultaneous detection of multiple metabolites from the same set of samples. The wide assay window allows detection of very small changes in metabolite concentration, an advantage over conventional fluorescent and colorimetric detection methods. The assays adapt to various formats (96, 384-well plates) and require no instrumentation other than a luminometer.
The assays can be used to measure the levels of critical metabolites and to monitor the effects of inhibitors and activators of glycolysis. The shift from oxidative phosphorylation to glycolysis can be monitored by measuring lactate production and secretion as well as glucose consumption from the same set of samples. The assays easily multiplex with cell viability assays enabling analysis of drug effects on cell proliferation, cell metabolism and mitochondria function.
Citation Format: Mary Sobol, Jolanta Vidugiriene, Donna Leippe, Gediminas Vidugiris, Wenhui Zhou, James Cali. Metabolite assays to illuminate cellular energy networks. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1211. doi:10.1158/1538-7445.AM2015-1211
Abstract 1220: MDM2 promotes p53-independent breast cancer metastatic phenotypes
Authors: Gao Chong , Kundu Nandini , Bargonetti Jill 
Published: August 2015

Abstract
Many human cancers overexpress the E3 ubiquitin ligase MDM2. The cancer genome atlas (TCGA) found that increased MDM2 expression is one of four gene pathways that correlate with all breast cancer subtypes. An increasing body of literature strongly suggests that overexpression of MDM2 can drive breast oncogenesis in the presence of mutant p53 (mtp53), as well as through molecular pathways that do not require wild-type p53 degradation. Therefore, the canonical wild-type p53 degradation pathway by MDM2 is clearly not the only important pathway. MDM2 expression has been shown to influence breast cancer cell invasion and metastasis. One potential mechanism for this is through the activation of SLUG (a transcription repressor of E-Cadherin) and the process of E-cadherin degradation. To investigate the p53-independent influence of MDM2 we engineered human breast cancer clones with inducible shRNA to knockdown MDM2 in MDA-MB-231 (mtp53 R280K), T47D (mtp53 L194F) and MCF-7 (wild-type p53) cells. Wound healing assays showed that MDM2 knockdown in all three cell lines reduced cell migratory capacity without altering p53 protein levels. In T47D.shmdm2 cells, a decrease in SLUG expression was observed with MDM2 knockdown indicating SLUG is a p53-independent target of MDM2. The knockdown of MDM2 also decreased slug and mmp9 mRNA detected by quantitative RT-PCR. To determine the biological significance of MDM2 in a system to recapitulate breast cancer architecture we examined T47D.shmdm2 cells in a 3-dimensional (3D) cell culture model. Importantly, the depletion of MDM2 in T47D.shmdm2 cells grown in 3D culture promoted luminal clearance. Our findings suggest that MDM2 can influence SLUG and MMP9 levels associated with breast cancer metastasis through a mechanism that does not require degradation of wild-type p53. Moreover our work suggests that MDM2 may promote changes to breast cancer architecture that drive metastasis.
Acknowledgements. This work was supported by a grant to JB from the Breast Cancer Research Foundation.
Citation Format: Chong Gao, Nandini Kundu, Jill Bargonetti. MDM2 promotes p53-independent breast cancer metastatic phenotypes. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1220. doi:10.1158/1538-7445.AM2015-1220