bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2520: TAK-733, a novel MEK1/2 inhibitor, inhibits tumor angiogenesis without affecting vascular permeability
Authors: Akira Hori , Tsuchiya Shuntarou , Kakoi Yuichi , Mizutani Akio , Vincent Patrick 
Published: February 2011

Abstract
TAK-733 is a highly potent, selective, and non-ATP competitive inhibitor of mitogen-activated protein kinase kinase 1/2 (MEK1/2). In preclinical studies, TAK-733 exhibits potent antitumor activity against a variety of human tumor xenografts, but does not affect vascular permeability. The effects of TAK-733 on tumor angiogenesis were investigated in this preclinical study. TAK-733 completely inhibited the vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)-induced ERK phosphorylation in human umbilical vein endothelial cells (HUVECs) at 100 nM. TAK-733 also inhibited VEGF- and FGF-induced HUVEC proliferation (IC50 = 1.3 and 18 nM, respectively) and HUVEC tube formation cocultured with fibroblasts (IC50 = 0.49 and 0.34 nM, respectively). The anti-angiogenic effect of TAK-733 in vivo was evaluated in a Matrigel plug assay following daily oral administration of TAK-733 at 3 and 10 mg/kg for 7 days. TAK-733 significantly inhibited the bFGF-induced hemoglobin accumulation in Matrigel plugs. TAK-733 significantly suppressed micro vessel density, examined by immunostaining with an anti-CD31 (PECAM) antibody, in human lung carcinoma A549 xenografts 4 and 7 days after initial treatment of 3 and 10 mg/kg. This was associated with significant antitumor effects in these xenograft models. These results indicate that potent anti-angiogenic activity of TAK-733 contributes to the anti-tumor activities. Angiogenesis inhibitors that modulate VEGF or VEGFR have all displayed hypertensive toxicity. Lack of nitric oxide production as a result of VEGF inhibition produces vasoconstriction and decreased vascular permeability leading to hypertensive effects (Fukumura et. al, PNAS 98, p2604, 2001). The effects of TAK-733 and VEGFR tyrosine kinase inhibitor on tumor interstitial fluid pressure (IFP) were examined by using wick-in-needle method in A549 xenograft model. TAK-733 did not affect the tumor IFP 6 hours after single administration of 30 mg/kg, while the VEGFR tyrosine kinase inhibitor significantly decreased it at 60 mg/kg. Consistent with the effects on tumor IFP, TAK-733 did show change of the vascular permeability unlike the VEGFR tyrosine kinase inhibitor by means of the Evans blue extravasation technique in A549 xenograft model. Although TAK-733 shows potent antiangiogenic effect, these profiles of TAK-733 on vascular permeability would not be expected to induce the typical toxicity of VEGF inhibitor.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2520.
Abstract 2523: BRAF is not the only predictor of sensitivity in vitro and in vivo for TAK-733, a selective potent inhibitor of MEK1/2
Authors: O'Connell Shawn M. , Tsuchiya Shuntarou , Shi Lihong , Fabrey Robyn , Zhang Lilly , Dong Qing , Hori Akira , Vincent Patrick 
Published: February 2011

Abstract
MEK has emerged as an attractive target for pharmacologic intervention in cancer as the RAS/RAF/MEK/ERK cascade has proven to play a central role in the signaling required for cellular transformation and proliferation. Inhibition of MEK impacts a diverse array of additional cellular events, including differentiation, apoptosis, and angiogenesis. Identification of an orally active inhibitor of MEK which shows significant efficacy as a mono-therapy for the treatment of cancer would be ideal. TAK-733 is a potent non-ATP competitive allosteric inhibitor of MEK1/2 with IC50 values for the inhibition of MEK1/2 signaling of 2-5 nM at high (400 μM) and low (10 μM) ATP concentrations. TAK-733 was selective for mammalian MEK-1/2 in a panel of 71 enzyme assays. These included phosphodiesterases, 17 kinases (IC50 values &gt;25 µM), cyclases, and 14 other enzymes (IC50 values &gt;10 µM). Using an MTS (a tetrazolium salt) viability assay, TAK-733 was found to inhibit proliferation of numerous cancer cell lines expressing BRAF, KRAS, NRAS, PI3K, MEK1, EGFR mutations, cMET amplification, and wild type MAPK signaling with EC50 values ranging from 2 to 90 nM. A quantitative PK/PD relationship was established in the HT-29 human colorectal carcinoma murine xenograft model. This model showed a range of PD activity (ie, from no effect to maximum effect) which correlated well with tumor and plasma TAK-733 concentrations. The relationship between TAK-733 plasma concentration (PK) and target modulation (PD) was evaluated by modeling percent inhibition of tumor ERK phosphorylation and plasma TAK-733 concentrations at 4 hours postdose, using an inhibitory sigmoidal concentration response model. The plasma EC50 was calculated to be 45 ng/mL. TAK-733 exhibited broad-spectrum in vivo antitumor activity against a panel of human tumor xenografts with and without mutations in the MAPK pathway. These xenografts were of colon (COLO 205/ BRAF V600E and HT-29/ BRAF V600E/PI3K P449T), NSCLC (A549/ KRAS G12S, PC-9/ EGFRΔE746-A750, NCI-H1975/ EGFR L858R T790M), AML (HL-60/ NRAS), cholangio-carcinoma (EGI-1/ KRAS G12D) and melanoma (A375/ BRAF V600E) origins. A high incidence of tumor regressions was observed in this collection of cancer models. Collectively, preclinical pharmacology studies indicate TAK-733 is a specific inhibitor of MEK with the potential to significantly impair growth of a large number of cancers relying on the MAPK signaling cascade for growth and survival. and provides a strong rationale for study of this agent in clinical trials.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2523.
Abstract 253: Identification of novel EWSR1-CREB3L1 fusion transcript in small cell osteosarcoma
Authors: Debelenko Larisa V. , Shivakumar Bangalore R. , McGregor Lisa , Dorfman Howard D. , Raimondi Susana 
Published: February 2011

Abstract
Small cell osteosarcoma (SCOS) is an aggressive variant of osteogenic sarcoma that has some morphologic similarities to Ewing sarcoma. Little is known about the molecular biology of SCOS and no specific molecular abnormality has been reported in this tumor subtype to date. We performed a molecular biology study of a SCOS that was positive for the Ewing sarcoma breakpoint region 1 (EWSR1) gene rearrangement by interphase fluorescent in situ hybridization and negative for EWSR1-FLI1, EWSR1-ERG and EWSR1-WT1 fusion transcripts by reverse transcriptase polymerase chain reaction. Rapid amplification of cDNA ends with the EWSR1 gene-specific forward primers revealed the exon 6 of the cAMP responsive element binding protein 3-like 1 gene (CREB3L1) fused in-frame to the EWSR1 exon 11, consistent with the EWSR1-CREB2L1 fusion transcript expressed in tumor tissue. The corresponding chimeric gene in the tumor DNA was confirmed by amplification and subsequent sequencing of the genetic breakpoint located in introns 11 and 5 of the EWSR1 and CREB3L1, respectively. No reciprocal fusion transcript or gene could be amplified in the tumor. An approximately 68 kDa product detected in tumor tissue lysate by immunoblotting with the CREB3L1 carboxyterminal antibody was consistent with a 656 aa predicted chimeric protein. This is the first report of a fusion transcript in osteosarcoma that demonstrates the relation of molecular mechanism of SCOS subtype to that of Ewing sarcoma. The 3'end fusion partner and the inferred structure of the new EWSR1-CREB3L1 chimera, however, are different from those underlying Ewing sarcoma, predicting different functions of the novel fusion product.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 253.
Abstract 2532: In vitro cytotoxicity, and pharmacokinetics, tissue distribution, and metabolism of the protein kinase D inhibitors, kb-NB142-70 and kb-NB184-43, in mice bearing human cancer xenografts
Authors: Jianxia Guo , Parise Robert A. , Beumer Jan H. , Clausen Dana M. , Egorin Merrill J. , Lazo John S. , Wipf Peter , Wang Q. Jane , Eiseman Julie L. 
Published: February 2011

Abstract
Introduction: Protein kinase D (PKD) contributes to prostate and pancreatic cancer cell growth and survival. Therefore, PKD inhibitors are potential anticancer agents. kb-NB142-70 (NB142) and its methoxy-analogue, kb-NB184-43 (NB184), inhibit PKD in vitro.
Methods: The in vitro inhibition of growth of Pc 3 prostate cells and Panc-1 pancreatic cells by NB142 and NB184 was evaluated by MTT assay. We evaluated the pharmacokinetics of NB142 in SCID mice bearing Pc 3 human prostate cancer xenografts and of NB184 in SCID mice bearing Panc-1 cancer xenografts. NB142 or NB184 was administered i.v. at 25 mg/kg. Three mice per time point were euthanized at 5, 15, 30, 60, 120, 240, 360, 960 and 1440 min after dosing. Plasma, tissues and urine were collected, and concentrations of NB142 and NB184 were quantitated by HPLC-UV. Metabolites in plasma and urine were characterized by LC-MS/MS.
Results: The IC50s of NB142 and NB184 were 21.0 µM and 24.3 µM, respectively, against Pc 3; and 33.7 µM and 27.4 µM, respectively, against Panc-1. After dosing NB142, the plasma NB142 Cmax (at 5 min) was 36.9 µM, and the NB142 Pc 3 tumor Cmax was 11.8 µM. NB142 was not detected in plasma, liver, kidney, spleen, or heart beyond 30 min. After dosing NB184, the plasma, liver, and kidney NB184 Cmax, (at 5 min) were 61.9 µM, 42.6 µM, and 94.8 µM, respectively. The NB184 Cmax in Panc-1 tumors (at 15 min) was 8.0 µM. Areas under the concentration vs time curves of NB142 and NB184 in plasma were 409 and 782 nmol·min/ml, respectively. The plasma half-lives of NB142 and NB184 were 6 and 14 min, respectively. The major metabolite of NB142 was a glucuronide. Between 0-6 h, urinary excretion of NB142 and metabolites accounted for 2.5 % and 7.6%, respectively, of the administered dose. NB184 underwent oxidation and glucuronidation, and more than 10 metabolites were observed. NB142 was not detected in plasma after administration of NB184. Between 0-16 h, urinary excretion of NB184 and metabolites accounted for 2.1 % and 5.3%, respectively, of the administered dose.
Conclusions: Although the inclusion of a methoxy group in NB184 increased its plasma half-life approximately 3-fold compared to NB142, metabolic and urinary elimination of both compounds was relatively rapid. Peak tumor concentrations of NB142 and NB184 were of the same order of magnitude as the concentrations required in vitro for cytotoxicity.
(Support: P30-CA47904 and P01-CA078039)
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2532.
Abstract 2546: IGR-1R pathway role in oxaliplatin acquired resistance in colon cancer cell lines and its regulation by MMP-7 and IGFBP-2
Authors: Codony-Servat Jordi , Martínez-Cardús Anna , Marín-Aguilera Mercedes , García-Albéniz Xabier , Augé Josep Maria , Gallego Rosa , Pineda Estela , Ginés Alba , Abad Albert , Gascón Pere , Martínez-Balibrea Eva , Maurel Joan 
Published: February 2011

Abstract
Background: IGF-1R pathway and MMPs have been implicated in chemotherapy resistance in colorectal cancer. This receptor could be activated by IGF1 or IGF2 ligands, which bioavailability is regulated by IGFBPs, what could be degraded by MMPs. Objectives: To determine the basal levels and the modulation by chemotherapy (oxaliplatin), of the levels of IGF-1R, IGFBPs and MMPs in colon cancer cell lines (HT-29 (IC50: 6,46 uM) and LoVo (IC50: 0,25 uM) and their oxaliplatin resistant cell lines (HT-29OXAR3 (IC50: 29,86 uM) and LoVOXAR3 (2,45 uM)). To analyze the differences in apoptosis between parental and oxaliplatin resistant cell lines when are treated with oxaliplatin. Methods: MMP-7 levels were analyzed by RT-PCR, Western Blot and ELISA. Levels of IGF-1R, activated IGF-1R, IGFBP-2, and IGFBP-3 were determined by Western Blot and/or ELISA. Differences in cell viability were analyzed by MTT, and intrinsic (bcl-2, bax) and extrinsic (Fas, caspase 8) apoptotic pathways, by Western Blot, in parental and oxaliplatin resistant cell lines with dose-response experiments. Results: LoVo and LoVOXAR3 express low levels of MMP-7 and IGF-1R and high levels of IGFBP-2 compared with HT-29 and HT-29OXAR3. In addition, HT-29OXAR3 cells showed increased MMP-7, IGF-1R and pIGF-1R levels compared with HT-29 and lower levels of IGFBP-2. We also observed after oxaliplatin treatment in HT-29 cells a decrement of IGF-1R and IGFBP-2 and an increment in MMP-7 levels. In apoptotic experiments, FAS and caspase 8 (extrinsic pathway) were induced by oxaliplatin in LOVO (0.2 uM) and in LOVOXAR3 (20uM). Bcl-2 and bax (intrinsic pathway) were induced or decremented respectively in LOVO (2uM) and LOVOXAR3 (20uM). No changes in these extrinsic apoptotic proteins were observed in HT29 and HT29OXR3. In HT-29OXAR3 cell line higher doses of oxaliplatin (20 uM instead of 2uM) were needed to induce intrinsic apoptotic (bax) and antiapoptotic proteins (bcl-2) Conclusions: IGF-1R signalling is implicated in acquired oxaliplatin resistance and it is regulated by MMP-7 and IGFBP-2 in a different manner in our two models of cancer colon cell lines. The two apoptotic pathways were implicated in LoVo cell lines, but only intrinsic pathway regulates apoptosis in HT-29 cell lines.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2546.
Abstract 2552: Loss of MiTF sensitizes melanoma cells to chemotherapy due in part to reduced LAMP1 accumulation
Authors: Liu Feng , Hwe Christopher , Chu Raymond , Meyskens Frank 
Published: February 2011

Abstract
MiTF (microphthalmia-associated transcription factor) was reported to be amplified in about 20% of melanoma samples and has an oncogenic property. However it is also known that in a subset of melanoma cell lines and tissues MiTF accumulation level is much reduced as compared to normal human melanocytes. Here we confirmed this observation in both melanoma cell lines and tissues. We found that MiTF accumulation level was independent of BRAF mutations status. Loss of MiTF was found at both mRNA and protein levels in melanoma cell lines. Interestingly, melanoma cell lines with MiTF accumulation showed higher resistance to multiple drug treatment. Further investigation revealed that lysosomal-associated membrane protein 1 (LAMP1) levels are tightly associated with MiTF accumulation and is a direct downstream transcription target for MiTF. Over-expression of LAMP1 in melanoma cells conferred drug resistance while knock-down LAMP1 by shRNA sensitized melanoma cells to chemotherapy drugs. Our studies revealed a novel transcription target for MiTF and a novel drug resistance mechanism that is mediated by MiTF, which may be associated with their intrinsic exocytosis function of melanoma cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2552.
Abstract 2555: Class I β-tubulin mutations induce resistance to microtubule destabilizer through stabilization of the microtubule networks
Authors: Cheung Chun Hei Antonio , Wu Su-Ying , Chang Chi-Yen , Hsieh Hsing-Pang , Chang Jang-Yang 
Published: February 2011

Abstract
Anti-mitotic compounds such as vincristine, colchicine and paclitaxel have been developed to target microtubules in cancers. Although these compounds have been shown successful in treating various cancers, development of drug-resistance cells limits their efficacies in clinical situations. To determine possible mechanisms that are responsible for drug-resistance to microtubule de-stabilizers, KB-derived drug-resistant KB-L30 cells were developed. KB-L30 cells showed resistance to various microtubule de-stabilizers through multiple-drug resistant (MDR)-independent mechanisms. In contrast, this specific clone showed hyper-sensitized to the microtubule-stabilizer, paclitaxel. Single-strand conformation polymorphism (SSCP) analysis, RT-PCR and DNA sequencing revealed no mutation on exon 1, 2 and 3 of βI-tubulin. Surprisingly, six novel mutation sites were found present in the exon 4 of βI-tubulin gene. Transfection of the βI-tubulin-mutated construct into KB cells demonstrated that these mutations could alter its sensitivity to the microtubule de-stabilizer, BPR0L075, in vitro. On the other hand, western blot analysis revealed increased microtubule assembly and reduced microtubule dynamic instability in KB-L30 cells when compared to its parental KB cells. In addition, computational modeling indicated that a direct relationship exists between tubulin mutations and alteration in the microtubule assembly and dynamic instability in KB-L30 cells. In conclusion, we showed for the first time that mutations in exon 4 of βI-tubulin induced resistance to microtubule de-stabilizers and hyper-sensitivity to microtubule stabilizer through an alteration in the microtubule assembly and dynamics in cancer cells. The results of this study have important implications for the development of future anti-mitotic compounds that are able to target microtubule mutation-induced drug-resistant cancer cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2555.
Abstract 2560: Cyclin-dependent kinase inhibitor SU9516 enhances sensitivity to methotrexate in human T-cell Jurkat cells
Authors: Uchiyama Hitoji , Sowa Yoshihiro , Wakada Miki , Yogosawa Mayumi , Nakanishi Ryoko , Horinaka Mano , Shimazaki Chihiro , Taniwaki Masafumi , Sakai Toshiyuki 
Published: February 2011

Abstract
Methotrexate (MTX), a classical anti-folate drug, has been used to treat various hematological malignancies. Since MTX prevents tumor cells from proliferating by inhibiting dihydrofolate reductase (DHFR), DHFR expression is a key determinant of resistance to MTX in malignant hematological tumor cells. In fact, it is well known that elevated expression of DHFR is a direct factor in clinical therapeutic resistance to MTX. First of all, we demonstrated that the sensitivity to MTX was enhanced by the knockdown of the DHFR gene in human T-cell leukemia cell. Concretely, the anti-proliferative effect of MTX was significantly enhanced by the knockdown of DHFR expression by siRNA in Jurkat cells. Therefore, a novel strategy down-regulating DHFR expression seems promising for enhancing sensitivity to MTX. On the other hand, the expression of DHFR is controlled by the transcription factor E2F. Unphosphorylated retinoblastoma (RB) protein is known to bind and inhibit the E2F. Cyclin-dependent kinase (CDK) inhibitors activate RB function and inhibit E2F function. Then we tested whether CDK inhibitor acted as a suppressor of DHFR through the E2F site of the promoter. We found that SU9516, a cyclin-dependent kinase inhibitor, decreased phospho-RB (Ser780) and reduced the expression of both DHFR mRNA and protein, and the DHFR promoter activity was attenuated by SU9516 dependent on the E2F site. Furthermore, pretreatment with SU9516 significantly enhanced sensitivity to MTX in a colony formation assay. To expand these results in Jurkat cells, we have also examined other leukemic cell lines (human T cell leukemia CCRF-CEM cells and human erythroleukemia K562 cells) and confirmed that SU9516 also repressed the levels of DHFR protein and phopho-RB (Ser780), and enhanced the cytotoxicity of MTX in a colony formation assay in both of cell lines. To expand these results by SU9516, we have examined other CDK inhibitors (Purvalanol A and Cdk4/6 Inhibitor IV) and confirmed that these agents also decreased DHFR protein and phospho-RB (Ser780), and enhanced the cytotoxicity of MTX in a colony formation assay in Jurkat cells as similar as SU9516. Based on these findings, we conclude that a combination of cyclin-dependent kinase inhibitors and MTX may be useful for overcoming resistance to MTX.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2560.
Abstract 2561: Genistein restored chemosensitivity to oxaliplatin in gemcitabine resistant pancreatic cancer cells in vitro and in orthotopic xenograft mouse model
Authors: Banerjee Sanjeev , Wang Zhiwei , Azmi Asfar , Kong Dejuan , Ahmad Aamir , Sarkar Fazlul H. 
Published: February 2011

Abstract
Chemotherapy remains the main stay for pancreatic cancer (PC) treatment with Gemcitabine showing modest benefit. Oxaliplatin has been used as a second line therapy in combination with gemcitabine but the beneficial effect was mitigated due to ‘off target’ toxicity as well as due to de novo and acquired resistance. However, effort toward restoration of sensitivity of PC cells to low concentrations of oxaliplatin is an emerging area of research. Here we report our in vitro and in vivo preclinical evidence in support of chemo-sensitization of gemcitabine resistant PC cells (Panc-1, Panc-28 and MiaPaCa-2) by a non-toxic chemopreventive agent known as genistein. Genistein pretreatment of PC cells exhibiting moderate (Panc-1, Panc-28) to less gemcitabine sensitivity (MiaPaCa-2) using low concentrations of oxaliplatin resulted in a significant reduction in the viability of cells concomitant with increased apoptosis (p&lt;0.01) occurred in a synergistic manner as assessed by colony formation experiments. Since drug-resistance of PC is allegedly linked with constitutive activation of NF-κB as well as further activation by conventional chemotherapeutic agents including oxaliplatin, our mechanistic showed that inactivation of NF-κB by genistein prior to treatment of cell with oxaliplatin is required for the killing of drug resistant cells, which indeed was consistent with the down- regulation of NF-κB and its downstream anti-apoptotic genes (Bcl-2 XIAP's, survivin) leading to the appearance of cleaved PARP and induction of apoptosis. Most importantly, our in vivo experiments using orthotopic mouse model of pancreatic cancer showed significant reduction in tumor size (p&lt;0.01) and reduction of locoregional lymph node metastasis when genistein was given in combination with oxaliplatin compared to monotherapy. These results were also consistent with inactivation of NF-κB and the down-regulation of NF-kB downstream targets (Bcl-xL, survivin and XIAP), all of which was consistent with in vitro findings. Both in vitro as well as in vivo findings were also consistent with immunohistochemistry findings on proliferation marker (Ki-67), apoptosis (TUNEL) and phospho p-65 immunostaining. From these results, we conclude that our results provide strong in vivo evidence in support of our hypothesis showing that genistein sensitizes drug-resistant pancreatic cancer to oxaliplatin, which is mechanistically linked with inactivation of NF-κB, which could be a useful strategy for the treatment of PC in the future.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2561.