bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract A290: Kinase activity in vemurafenib-sensitive and resistant BRAF-mutant melanoma cells.
Authors: Risberg Karianne , Røe Kathrine , Ree Anne H. 
Published: August 2014

Abstract
In advanced melanoma, therapeutic targeting of the MAPK signaling pathway with vemurafenib, a potent inhibitor of the BRAFV600E-encoded signaling molecule, commonly results in rapid and substantial tumor responses, but limited durability of the clinical benefit. Therapy resistance inevitably develops either via reactivation of the MAPK (RAF/MEK/ERK) pathway or via bypass mechanisms, such as PI3K/AKT/mTOR-mediated signaling; however, no one mechanism of resistance has been consistently observed on disease progression. In preclinical models, dual inhibition of the MAPK pathway (by MEK inhibitors) and PI3K/AKT/mTOR signaling has shown to be an effective strategy at overcoming this type of resistance, and clinical studies are ongoing.
Hypothesizing that different patterns of composite kinase signaling activities in BRAF-mutant melanoma may reflect the response of vemurafenib and other inhibitors (e.g. MEK and AKT inhibitors), we analyzed the tyrosine kinase activity profiles generated by a vemurafenib sensitive melanoma cell line to a vemurafenib resistant melanoma cell line, both harbouring the BRAFV600E mutation. Of note, both cell lines were derived from metastatic tumor biopsies sampled from a patient with disseminated melanoma disease, participating in the BRIM-2 trial, before commencing vemurafenib treatment and at six weeks on treatment. The patient experienced partial therapy response lasting for 15 weeks before disease progression.
Interestingly, overall kinase activity was lower in the vemurafenib-resistant cell line, and the phosphorylation levels of a number of array substrates representing mediators of the MAPK and PI3K/AKT/mTOR signaling pathways were different in the cell lines sensitive and resistant to vemurafenib. Whilst treatment with vemurafenib, the MEK inhibitor U0126 and the AKT inhibitor M2266 resulted in limited effect in the vemurafenib-resistant cells, the drugs caused decreased in vitro cell viability in the vemurafenib-sensitive cell line.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A290.
Citation Format: Karianne Risberg, Kathrine Røe, Anne H. Ree. Kinase activity in vemurafenib-sensitive and resistant BRAF-mutant melanoma cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A290.
Abstract A293: Novel allosteric small molecule FAK inhibitors effectively inhibit cancer cell viability and clonogenicity.
Authors: Golubovskaya Vita M. , Lebedyeva Iryna , Ho Baotran , Yemma Michael , Singh Neha , Arcara Makayla , Ostrov David , Katritzky Alan , Cance William 
Published: August 2014

Abstract
Introduction: Focal Adhesion Kinase is overexpressed and activated in many types of tumors and plays an important role in survival. Recently, we developed FAK inhibitor, 1,2,4,5-benzenetetraamine tetrahydrochloride, called Y15, which inhibited FAK autophosphorylation and blocked breast, neuroblastoma, pancreatic, brain and colon cancer tumor growth. Experimental procedures: In this study, eight novel chemical derivatives of Y15, called Y15-I1-I8 were synthesized and tested in a panel of different cancer cell lines. To test the effect on FAK autophosphorylation we performed in vitro ADP-Glo kinase assay with recombinant FAK protein and Western blotting with Y397-FAK antibody. To test the in vivo effect of FAK inhibitors we performed MTT assay, AnnexinV apoptosis and clonogenicity assays in different cancer cell lines. As a control, we used Novartis NVP-TAE-226 FAK inhibitor. Results: By in vitro kinase assay Y15-I1-8 derivatives effectively inhibited FAK autophosphorylation with recombinant purified full length FAK protein. Western blotting demonstrated decreased Y397-FAK and Y418-Src phosphorylation in many cancer cell lines. In addition, these novel FAK inhibitors demonstrated decreased viability, increased apoptosis and inhibition of clonogenicity in lung, melanoma, glioblastoma, pancreatic, breast, and colon cancer cell lines. Some of the novel derivatives were better than parental Y15 inhibitor. Conclusion: Targeting FAK autophosphorylation with novel allosteric derivatives of Y15 is an effective therapy approach.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A293.
Citation Format: Vita M. Golubovskaya, Iryna Lebedyeva, Baotran Ho, Michael Yemma, Neha Singh, Makayla Arcara, David Ostrov, Alan Katritzky, William Cance. Novel allosteric small molecule FAK inhibitors effectively inhibit cancer cell viability and clonogenicity. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A293.
Abstract A45: Biomarker assay performance: The impact of the fixation method.
Authors: Badola Sunita 
Published: August 2014

Abstract
Recent advances in genomic technologies dramatically illustrated the complexity of cancer genomes. These discoveries provide new opportunities for drug development and ultimately better patient care by informing treatment choice decisions in addition to the insights into the mechanisms of response or relapse to therapy. Formalin fixed paraffin embedded (FFPE) tumor tissue specimens routinely collected in Oncology diagnostic practice present a number of challenges for performing molecular biomarker assays. These problems include chemical cross-linking RNA/DNA and protein, nucleic acid fragmentation, and reduced amount of extractable material. We evaluated a new fixative agent (PAXgene, Qiagen) to assess whether an alternative fixation method could alleviate problems presented by the FFPE specimens.
PAXgene preservation method was compared to FFPE and frozen tissue utilizing tumor material from xenograft and primary tumor mouse models. We compared the performance of the following techniques across three types of specimens: allele-discrimination PCR, gene expression profiling by QPCR, CNV (copy number variation) analysis, and IHC. DNA and RNA based assay performance including allele-specific PCR, gene expression profiling and CNV demonstrated similar results between frozen and PAXgene tumor specimens. Comparison of transcriptional profiling data between all there type of specimens demonstrated artifacts likely related to the formalin fixation process. Additionally, DNA extracted from PAXgene specimens had higher amount of amplifiable templates compared to DNA extracted from the FFPE specimens. Tissue sections from PAXgene fixed specimens exhibited preserved tissue morphology of acceptable quality. We carried out pAKT and PTEN IHC assays in PAXgene and FFPE tissue sections. The results demonstrated superior performance for pAKT in the PAXgene format. In the case of PTEN IHC assay, the standard FFPE based IHC assay protocol required substantial modifications.
Collecting tumor biopsies in the PAXgene format provides an opportunity of conducting both molecular and tissue based biomarker assays from a single tumor specimen.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A45.
Citation Format: Sunita Badola. Biomarker assay performance: The impact of the fixation method. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A45.
Abstract A46: Expression of biomarkers in cervical intraepithelial neoplasia: Potential use screening.
Authors: Espinosa Garcia Ana Maria , Alfaro Ana , Palma Icela , Duran Marco , Beumen Jaime 
Published: August 2014

Abstract
Background: Persistent infection of high-risk Human papillomavirus (HPV) is recognized as a necessary cause of cervical cancer. Replication and viral integration into the cervix depend on the orderly expression of genes produced by viruses, by inducing the overexpression of multiple molecular proteins or biomarkers. Recently we have identified, 6 genes (CCNB2, CDC20, PRC1, SYCP2, NUSAP1 and CDKN3), which are proposed as universal molecular targets with a sensitivity and specificity close to100%, in cervical cancer.
Objective: evaluate the expression of these genes in cervical intraepithelial neoplasia (CIN).
Material and method: In this retrospective study we assessed the protein expression of 6 candidates tissue-based biomarkers (CCNB2, CDC20, PRC1, SYCP2, NUSAP1 and CDKN3) in a population based cohort of 200 cervical intraepithelial neoplasia that was collected over the period from 2008 to 2013. The tissue blocks were available and used for tissue microarray (TMA) construction, which included also samples of healthy epithelium. Quantitative RT-PCR was performed to validate expression of genes.
Results: The expression of these genes was observed in cervical intraepithelial neoplasia with different pattern shown previously in tumors, but in progression with the advance of the lesion, also shown by IH, evaluated samples, finding a correlation in both methods.
Conclusion: These results, may offer an additional strategy to current methods of the PAP, which is of greater use in our population. A test with greater sensitivity and specificity to identify lesions of high grade with more specificity can be a useful tool for cervical cancer screening.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A46.
Citation Format: Ana Maria Espinosa Garcia, Ana Alfaro, Icela Palma, Marco Duran, Jaime Beumen. Expression of biomarkers in cervical intraepithelial neoplasia: Potential use screening. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A46.
Abstract A49: Anti-DLL4 antibodies inhibit cancer stem cells in small cell lung cancer.
Authors: Strout Patrick , Korade Martin , Leow Ching Ching , Inigo Ivan , Thomas Suneetha , Hurt Elaine , Chesebrough Jon , Kamal Adeela , Cho Song 
Published: August 2014

Abstract
Small cell lung cancer (SCLC) is a very aggressive lung cancer with features suggesting enrichment in cancer stem cells (CSCs). Delta-like ligand 4 (DLL4) is a membrane bound ligand for Notch receptors critical for functional angiogenesis. Blocking DLL4 signaling increases the density of nonfunctional blood vessels and hypoxia of tumors, and thereby inhibits growth of tumor xenografts in mice. In addition, growing evidence implicates DLL4 Notch signaling pathway in the maintenance of CSCs. Here we investigated the anti-CSC activity of anti-DLL4 mAbs using in vitro and in vivo models of SCLC. Biochemical and flow cytometry analyses revealed that multiple SCLC cell lines express DLL4, and notably, SCLC spheres cultured under CSC-enriching conditions express higher levels of DLL4. Prior to determining the ability of anti-DLL4 mAb in inhibiting CSC in vivo, we first characterized the phenotype of CSCs in the SCLC cell line NCI-H69. NCI-H69 cells enriched for high CD133 expression were more tumorigenic and expressed higher levels of Nanog, Oct 3/4, and EZH2, which are genes crucial for maintenance of CSCs, than cells with low or negative levels of CD133. Moreover, RNA level of DLL4 was found to be three fold higher in CD133 high cells compared to CD133 low cells, suggesting that expression of DLL4 correlates with the CSC phenotype. In vivo, an anti-DLL4 antibody leads to inhibition of NCI-H69 tumor growth when the antibody is administered as a single agent, or in combination with cisplatin+etoposide or topotecan, chemotherapy agents commonly used in treatment of SCLC. To determine if blockade of DLL4 inhibits CSCs in vivo, NCI-H69 tumor xenografts treated with anti-DLL4 mAb were analyzed by flow cytometry. A subset of dissociated tumor cells expressed CD133 and DLL4, and a reduction of these populations of cells was observed in the anti-DLL4 antibody treated groups. In summary, these studies highlight that, in addition to vascular expression, (1) DLL4 is frequently expressed in SCLC cells, (2) DLL4 expression correlates with a CSC phenotype, and (3) that DLL4 blockade using an anti-DLL4 mAb results in inhibition of CSCs.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A49.
Citation Format: Patrick Strout, Martin Korade, Ching Ching Leow, Ivan Inigo, Suneetha Thomas, Elaine Hurt, Jon Chesebrough, Adeela Kamal, Song Cho. Anti-DLL4 antibodies inhibit cancer stem cells in small cell lung cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A49.
Abstract A53: Yin Yang 1 (YY1) regulates EMT via transcriptional activation of Snail and its overexpression in cancer stem cells (CSCs) in prostate cancer: Clinical significance.
Authors: Baritaki Stavroula , Rapozzi Valentina , Pelayo Rosana , Vega Mario , Huerta-Yepez Sara , Bonavida Benjamin 
Published: August 2014

Abstract
The epithelial to mesenchymal transition (EMT) is a cellular program that confers on neoplastic cells the biological traits needed to accomplish the metastatic cascade. The activation of EMT has been associated with the acquisition of stem cell traits, i.e., cancer stem cells (CSCs). CSCs have been reported to display EMT characteristics. Further, both EMT+ cells and CSCs exhibit a drug-resistant phenotype, hence, there is an urgent need to identify new therapeutic targets to eliminate both CSCs and EMT+ cells. Our previous findings and others’ demonstrated that the transcription factor Yin Yang 1 (YY1) regulates both the drug resistance and the metastasis-inducer Snail. Thus, we hypothesized that YY1 might be a multi-potential therapeutic target. We used human prostate carcinoma cell lines (PC3 EMT+ and LnCaP EMT-) as models. We demonstrate that overexpression of YY1 in LnCaP rendered the cells EMT+ (Snail+, E cadherin-, RKIP-). In contrast, treatment of PC3 with siRNA YY1 reverted the cells to the MET phenotype. Analysis by IHC in TMA of human prostate cancer tissues showed a positive correlation between the expression of Snail and YY1. Further, bioinformatic analysis corroborated the coexpression of Snail and YY1 in prostate cancer. Isolation of CSCs (CD44+, CD133+, CD24+) from PC3 cells exhibited overexpression of YY1 and other EMT markers. The inhibition of YY1 by various chemical inhibitors sensitized the resistant tumor cells to apoptosis to both chemo/immuno-cytotoxic drugs. These findings demonstrate that YY1 regulates EMT and drug resistance and is a prognostic biomarker. Further, we suggest that YY1 is a novel therapeutic target for the elimination of CSCs and metastasis in prostate cancer. Based on YY1 overexpression in many tumor types, we suggest that the present findings and their clinical significance may also be generalized to other cancers.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A53.
Citation Format: Stavroula Baritaki, Valentina Rapozzi, Rosana Pelayo, Mario Vega, Sara Huerta-Yepez, Benjamin Bonavida. Yin Yang 1 (YY1) regulates EMT via transcriptional activation of Snail and its overexpression in cancer stem cells (CSCs) in prostate cancer: Clinical significance. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A53.
Abstract A61: Characterization of KRAS-driven survival signaling networks via phosphoproteomics in lung cancer.
Authors: Kim Jae-Young , Welsh Eric A. , Fang Bin , Li Jiannong , Eschrich Steven A. , Koomen John , Haura Eric B. 
Published: August 2014

Abstract
KRAS mutations account for development of approximately 30% of lung adenocarcinomas, yet therapeutic options remain limited. To better understand KRAS-mediated survival mechanisms, we characterized phenotypic effects of KRAS knockdown as well as inhibition of its downstream effectors, including MEK, PI3K and RalB using 11 KRAS mutant non-small cell lung cancer cell lines. We observed only 2 cell lines with clear evidence of apoptosis after shRNA-mediated KRAS knockdown (H23 and Calu6). Interestingly, neither pharmacologic inhibition of MEK and PI3K nor knockdown of RalB fully recapitulated apoptotic phenotypes observed upon KRAS knockdown in these KRAS-sensitive lung cancer cells, suggesting that KRAS promotes survival through additional mechanisms. In order to identify novel KRAS survival pathways, we searched for altered phosphoproteins following shRNA-mediated KRAS knockdown using mass spectrometry (MS)-based phosphoproteomics. Stable isotope labeling by amino acids in cell culture (SILAC) was performed and phosphopeptides (p-Ser/Thr/Tyr) were purified using immobilized metal affinity chromatography (IMAC) resin, then analyzed by LC-MS/MS from sh-control or sh-KRAS-infected H23 lung adenocarcinoma cells. In total, we identified 2,878 phosphopeptides, corresponding to 1375 phosphoproteins, of which 210 phosphopeptides were upregulated and 204 phosphopeptides were downregulated after KRAS knockdown (cutoff: 1.5 fold). Using the altered phosphopeptides, we composed a cohesive network view to KRAS-regulated phosphoproteins (Kim et al. PNAS, 110;12414-12419), which revealed that KRAS regulates phosphorylation of cyclin-dependent kinase 1 and 2 (CDK1/2), and downstream mitotic targets. We also observed that KRAS knockdown activates adaptive resistance mechanisms via increased phosphorylation of RAF, PRAS40 and YAP1.
We are currently characterizing mitosis-related KRAS functions. Additionally, we are evaluating adaptive resistance mechanisms followed by loss of KRAS by analyzing KRAS-regulated phosphoproteome in cells harboring KRAS mutations but resistant to KRAS loss. These studies will provide better insights into KRAS-regulated survival signaling mechanisms and adaptive responses, which could offer rational co-targeting strategies. Updated work will be presented.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A61.
Citation Format: Jae-Young Kim, Eric A. Welsh, Bin Fang, Jiannong Li, Steven A. Eschrich, John Koomen, Eric B. Haura. Characterization of KRAS-driven survival signaling networks via phosphoproteomics in lung cancer. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A61.
Abstract A65: Characterization of CBP/p300-mediated butyrate-induced Wnt activity in colonic microadenoma cells.
Authors: Lazarova Darina L. , Chiaro Christopher , Wong Terrence , Bordonaro Michael 
Published: August 2014

Abstract
Objectives of the study: We wished to (a) compare gene expression profiles between a colonic microadenoma cell line (LT97) and a metastatic colorectal cancer (CRC) cell line (SW620) in the presence and absence of butyrate, (b) determine whether the sensitivity of LT97 cells to the growth suppressive effects of butyrate correlates to a high-fold upregulation of Wnt signaling in these cells, and (c) evaluate how modulation of CBP/p300-mediated Wnt signaling influences the ability of butyrate to upregulate Wnt activity in LT97 cells.
Methods: RNA was isolated from mock treated or butyrate treated LT97 microadenoma cells or SW620 metastatic CRC cells, followed by full human genome microarray analysis (Genus Biosystems). Colonic cells were transfected with Wnt reporter plasmids and treated with ICG-001 (which inhibits CBP-mediated Wnt activity) or co-transfected with a p300 expression vector or with an expression vector for KLF4, which inhibits CBP/p300 association with beta-catenin. Reporter assays, western blotting, and caspase assays were performed.
Results: We demonstrate that LT97 colonic microadenoma cells differ substantially in gene expression profiles compared to a metastatic CRC cell line (SW620), both in the presence and absence of butyrate. Further, we demonstrate that the high degree of sensitivity to the growth repressive effects of butyrate of LT97 cells correlates to a marked butyrate-induced hyperactivation of Wnt activity in that cell line. Further, the induction of Wnt activity by butyrate in LT97 cells is sensitive to treatment with ICG-001, overexpression of CBP or p300, or blockade of CBP/p300-mediated Wnt activity by the transcription factor KLF4.
Conclusions: Microadenoma cells, reflecting very early stage colonic neoplasia, are highly sensitive to the effects of butyrate on Wnt signaling, in a manner dependent upon CBP/p300. The findings generated from this study may lead to approaches to modulate the action of CBP/p300-mediated Wnt signaling transcriptional programs for therapeutic or chemopreventive effect.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A65.
Citation Format: Darina L. Lazarova, Christopher Chiaro, Terrence Wong, Michael Bordonaro. Characterization of CBP/p300-mediated butyrate-induced Wnt activity in colonic microadenoma cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A65.
Abstract A7: Minocycline regulates Hypoxia-Inducible Factor-1α expression through the induction of its degradation in ovarian cancer: in vitro and in vivo studies.
Authors: Ataie-Kachoie Parvin , Pourgholami Mohammad H. , Morris David L. 
Published: August 2014

Abstract
Experimental data suggest therapeutic advantage from selective disruption of the hypoxia response in cancer. We recently found that minocycline decreases tumor VEGF expression in ovarian tumor xenografts in mice and herein report a companion laboratory study to test if this effect was the result of hypoxia-inducible factor (HIF) inhibition. Human ovarian carcinoma cell lines (OVCAR-3, SKOV-3 and A2780) were treated with minocycline in normoxic and hypoxic conditions and also in the presence of hypoxia-mimetic agent deferoxamine in vitro. Two different in vivo experiments investigated miocycline effects after single dose (30 mg/kg, 4 or 24 h) or longer exposure in nude mice bearing OVCAR-3 xenografts. Treatment with minocycline inhibited the accumulation of HIF-1alpha in a concentration-dependent fashion in vitro. A similar effect of minocycline was seen in vivo. Furthermore, the inhibtory effect of minocycline on HIF-1alpha was found to be associated with a significant decrease in HIF-1alpha half-life showing that minocycline enhances degradtion of HIF-1alpha. These data warrents further evaluation of minocycline in the treatment of ovarian cancer.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A7.
Citation Format: Parvin Ataie-Kachoie, Mohammad H. Pourgholami, David L. Morris. Minocycline regulates Hypoxia-Inducible Factor-1α expression through the induction of its degradation in ovarian cancer: in vitro and in vivo studies. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A7.