bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1416: Long non-coding RNA GAN1 inhibits tumor progression by regulating the mir-26a/PTEN axis in lung cancer
Authors: Wang Rui-qi , Long Xiao-Ran , Zhang Mei-Yin , Chen Dong-Ni , Wen Zhe-Sheng , Mai Shi-Juan , Wang Hui-Yun 
Published: September 2020

Abstract
Background: Long non-coding RNA has been reported to be involved in many cancers. To explore the role of lncRNA in non-small-cell lung cancer (NSCLC), we detected lncRNA expression profile with a custom microarray and found that lnc-GAN1 was significantly downregulated in NSCLC. In this study, we aimed to explore the clinical significance, biological function and mechanism of lnc-GAN1 in NSCLC.
Methods: We firstly analyzed relationship between the lncGAN1 expression and clinical features and survival in NSCLC patients from two medical centers. We secondly examined the biological function of lncGAN1 in lung cancer cells with Cell viability, colony formation, migration, invasion, cell cycle and apoptosis assays. Then, we localized the distribution of lnc-GAN1 in lung cancer cells using RNA-FISH and analyzed the interaction between lnc-GAN1 and mir-26a-5p with Dual luciferase reporter and RNA immunoprecipitation (RIP) assay. Finally, the xenograft tumor model was employed to assess tumor suppression role of GAN1.
Results: lnc-GAN1 is notably downregulated in NSCLC tissues compared with normal lung tissues and associated with small tumor size, poor overall survival (OS) and disease-free survival in NSCLC patients. In vitro experiments, lnc-GAN1 expression suppresses proliferation, colony formation, cell cycle progression and apoptosis of lung cancer cells. lnc-GAN1 was mainly observed in cytoplasm of lung cancer cells with FISH and subcellular fraction analysis. RNA-seq analysis showed that the genes in cell cycle pathway were changed obviously when lnc-GAN1 was overexpressed. Mechanistically, lnc-GAN1 functions as a sponge of miR-26a-5p, which directly targets PTEN. Ectopically expressed lnc-GAN1 increased the mRNA and protein levels of PTEN and decreased miR-26a-5p level in lung cancer cells. Furthermore, mir-26a-5p mimics caused a decrease in both lnc-GAN1 and PTEN expressions while mir-26a-5p inhibitors led to an increase in both lnc-GAN1 and PTEN levels. Finally, In vivo experiments demonstrated that lnc-GAN1 could suppress tumor growth compared with control vector in nude mice.
Conclusions: our study discovered that lnc-GAN1 is significantly downregulated and associated with survival in NSCLC patients, and plays a tumor suppressor role by inhibiting miR-26a-5p and upregulating PTEN expression in lung cancer, which suggest that lnc-GAN1 is a potential prognostic biomarker and innovative therapeutic target in NSCLC.
Citation Format: Rui-qi Wang, Xiao-Ran Long, Mei-Yin Zhang, Dong-Ni Chen, Zhe-Sheng Wen, Shi-Juan Mai, Hui-Yun Wang. Long non-coding RNA GAN1 inhibits tumor progression by regulating the mir-26a/PTEN axis in lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1416.
Abstract 1436: Determination of antibody's specificity towards phosphorylated protein targets with automated in-capillary enzyme treatment and immunoassay
Authors: Taketa Daryl A. , Grant Rainer , Aydogdu Tufan 
Published: September 2020

Abstract
Protein phosphorylation is a reversible reaction that is integral in numerous signaling cascades. Characterization of signaling cascades has been largely detected by immunoblotting with phospho-specific antibodies, which may or may not have enough specificity or affinity. Currently, a separate lysate without any phosphatase inhibitors or a separate blot is needed to determine an antibody's specificity. Here we describe a simple assay that leverages automation and quantitation with capillary electrophoresis-based immunoassay (CEIA) to assess the specificity of these antibodies with a single lysate preparation. In this study, three lysate models are used: K562 ± TNFα treatment, 50 ng/mL phorbol myristate acetate (PMA) differentiated THP-1 ± 1 μg/mL lipopolysaccharide (LPS) treatment, and cytotoxic T lymphocytes (CTL) ± 10 ng/mL PMA and 500 ng/mL ionomycin treatment. K562 cell lysates are commercially purchased whereas THP-1 lysates are generated in-house. For CTL cells, whole blood cells from a single donor are isolated and expanded with commercially available kits. Expanded CTL cells are then stimulated with PMA and ionomycin for 15 minutes. Untreated and treated lysate samples are separated and captured to the inner lumen of the capillary wall with UV activated crosslink chemistry. Cross-linked proteins are treated with lambda phosphatase for 1 hour followed by the immunoassay to investigate the specificity of antibodies against phosphorylated protein targets respective to each activated pathway using either chemiluminescent or fluorescent detection. Preliminary data suggest phospho-specific signal decreased &gt;90% with no significant changes to the non-specific noise. The method described here eliminates the need for multiple lysate preparations or an additional blot to assess an antibody's specificity to a phosphorylated protein target.
Citation Format: Daryl A. Taketa, Rainer Grant, Tufan Aydogdu. Determination of antibody's specificity towards phosphorylated protein targets with automated in-capillary enzyme treatment and immunoassay [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1436.
Abstract 1453: Specific detection of mutant Ras oncoproteins in cell and tissue lysates by multiplex immunoassay
Authors: Hwang Joseph B. , Saporita Anthony , Xiao Qiang 
Published: September 2020

Abstract
Ras proteins (HRAS, KRAS, and NRAS) are small GTPases that function as molecular switches by alternating between inactive GDP-bound and active GTP-bound states. Ras-GTP activates downstream pathways, including MAPK, by binding to Raf kinase and phosphatidylinositol 3-kinase (PI3K) to promote cellular proliferation, survival, growth and differentiation. Oncogenic mutations in Ras are found in approximately 25% of human cancers. Furthermore, 90% of pancreatic cancers harbor KRAS mutations with 80% of KRAS mutations occurring at codon 12 with G12V and G12D being the most prevalent. The Ras-Raf-MEK pathway can also be activated downstream of Ras. BRAF is a well-established oncogene, with the BRAF V600E mutation being prevalent in melanoma. While targeting this BRAF oncoprotein has had therapeutic efficacy, treating cancers driven by Ras oncoproteins remains an urgent unmet clinical need. We developed a novel multiplex immunoassay capable of simultaneously detecting total Ras, RasG12V, RasG12D, phospho-MEK1 (S217/S221), phospho-BRAF (S446) and phospho-CRAF (S338) in a single well. This multiplex assay detects the H-Ras, N-Ras, and K-Ras isoforms for total Ras, RasG12D, and RasG12V. To confirm specific recognition of mutant Ras oncoproteins, this kit was able to detect the known KRAS G12V mutation found in the COR-L23 cell line and the NRAS G12D mutation found in THP-1 cell line, respectively. In addition, the response to BRAF inhibitors vemurafenib and dabrafenib was compared between MDA-MB-435S cells, which harbor the BRAF V600E mutation, and insensitive cell lines, using multiplex immunoassays. While COR-L23 and THP-1 cells were not responsive to vemurafenib and dabrafenib, MDA-MB-435S cells showed a dramatic reduction in pMEK1 and pERK, consistent with inhibition of mutant BRAF protein. We then analyzed commercially-sourced tissue lysates, using tumor and adjacent tissue from a patient with colon cancer harboring a KRAS G12D mutation. While multiplex protein analysis revealed a similar phosphorylation pattern for Raf and MEK proteins, the RasG12D oncoprotein was specifically observed in lysate from the tumor but not the adjacent tissue. Finally, exosomes were enriched from pancreatic cancer serum and tested using the Ras-Raf Oncoprotein panel. These exosomal preparations yielded detectable signal for Total Ras and phospho-BRAF. Our data demonstrate the utility of studying the effects of Ras oncoproteins by multiplex immunoassay in multiple sample types, including cell, tissue, and exosomal lysates.
Citation Format: Joseph B. Hwang, Anthony Saporita, Qiang Xiao. Specific detection of mutant Ras oncoproteins in cell and tissue lysates by multiplex immunoassay [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1453.
Abstract 1466: Molecular insights of resistin associated proliferation by inducing hypercholesterolemia in breast cancer
Authors: Deb Ankita , Bhat Manoj 
Published: September 2020

Abstract
Introduction: Breast cancer is the most prevalent cancer in women impacting ~2.1 million women each year (WHO, 2019). It is the second leading cause of death after lung cancer in women. In spite of several therapeutic advances, further improvement in prognosis and survivability is still desired. According to numerous studies, Resistin, a proinflammatory cytokine, has a positive correlation with breast tumor progression. It plays multifaceted roles in diabetes, obesity, cardiovascular diseases as well as in several cancers. Despite several epidemiological studies showing an increase in serum resistin level in breast cancer patients, its relevance is not clearly understood. In the present study, we have focused on the role of resistin in breast cancer and emphasized on metabolic alterations occurring in this process.
Methods: In vitro studies include long term colony formation and LDL estimation assays, immunoblotting, immunofluorescence . For in vivo studies, female Balb/C mice were used as isograft breast cancer model.
Results: In vitro studies show that resistin facilitates progression of breast cancer through activation of JAK-STAT pathway. Downstream targets, c-Myc and cdc25A are also regulated via Resistin. Inhibition of JAK-STAT pathway abrogates resistin induced growth in breast cancer cells. Moreover, resistin also promotes cholesterol accumulation thereby inducing a hypercholesterolemic environment. In vivo studies indicate that with increase in tumor progression, resistin as well as LDL levels are also elevated in serum.
Conclusions: In vitro as well as in vivo studies show that increase in serum resistin level is positively correlated with breast cancer progression and induction of hypercholesterolemia. Insights into resistin associated molecular events contributing to metabolic changes might facilitate progression of breast cancer. Targeting this pathway might contribute to a better therapeutic outcomes.
Citation Format: Ankita Deb, Manoj Bhat. Molecular insights of resistin associated proliferation by inducing hypercholesterolemia in breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1466.
Abstract 1470: UGP2 is a critical regulator of protein glycosylation in pancreatic cancer
Authors: Wolfe Andrew L. , Galeas Jacqueline , Toska Eneda , Zhou Qing , Ku Angel , Koche Richard P. , Bandyopadhyay Sourav , Lebrilla Carlito B. , Scaltriti Maurizio , McCormick Frank , Kim Sung Eun 
Published: September 2020

Abstract
UDP-glucose pyrophosphorylase 2 (UGP2) is a lynchpin metabolic enzyme that rests at the convergence of multiple metabolic pathways regulating both glycogen synthesis and glycosylation modifications. Here we elucidated the essential role of UGP2-mediated glycosylation in the maintenance of KRAS-driven cancer using both in vitro and in vivo models. A key effector of KRAS oncogenic function is the transcription factor YAP. We identified the YAP/TEAD transcription factor complex as a major regulator of UGP2 mRNA expression and enzymatic activity using genomic perturbations, correlative protein immunohistochemistry in PDAC samples, and ChIP-seq and targeted ChIP-qPCR at the UGP2 promoter. Loss of YAP or UGP2 leads to a decrease in glycogen and defects in key glycosylation targets such as EGFR. In murine xenograft models using pancreatic cancer lines, knockdown of UGP2 prevented tumor growth and decreased expression of the critical glycosylation target EGFR. This essential role for UGP2 in cancer maintenance may lead to new avenues of therapy in otherwise intractable KRAS-driven cancers.
Citation Format: Andrew L. Wolfe, Jacqueline Galeas, Eneda Toska, Qing Zhou, Angel Ku, Richard P. Koche, Sourav Bandyopadhyay, Carlito B. Lebrilla, Maurizio Scaltriti, Frank McCormick, Sung Eun Kim. UGP2 is a critical regulator of protein glycosylation in pancreatic cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1470.
Abstract 1477: LRG1 blockade normalizes tumor vasculature and improves efficacy of chemotherapy
Authors: Pilotti Camilla , O'Connor Marie N , Kallenberg David , Dowsett Laura , George Jestin , Moss Stephen E. , Greenwood John 
Published: September 2020

Abstract
In cancer blood vessels are dysfunctional, poorly perfused and leaky. Malfunctioning vessels contribute to the pro-oncogenic environment and limit the efficacy of current systemically administered drugs. Normalizing the tumor vasculature to improve vessel permeability, reduce hypoxia and vascular leakage and enhance drug delivery, has become an experimental objective in cancer research. This study was carried out to investigate the effect of blocking the secreted glycoprotein leucine-rich alpha-2-glycoprotein 1 (LRG1) on tumor vascular function, and evaluate the impact it has on the efficacy of the common standard of care chemotherapeutic drug cisplatin. Under normal conditions LRG1 is mainly expressed in the liver but also in other tissues such as bone marrow and immune cells. LRG1 has been described in multiple reports to be a serum prognostic biomarker in several cancers, for example lung, prostate, colorectal and breast. LRG1 promotes dysfunctional vessel growth by disrupting TGFβ signaling. We demonstrate that in Lrg1-/- mice and following treatment with a LRG1 function-blocking antibody (15C4) tumor growth was inhibited. In addition, we show using RNAscope that following subcutaneous grafting of the B16F0 and LL2 tumor cell lines in mice, Lrg1 is induced in tumor endothelial cells. Despite having no effect on total vessel area, the density was decreased upon LRG1 blockade, with the persisting larger vessels exhibiting improved vessel structure as evidenced by increased pericyte and basement membrane endothelial cell coverage. Better mural cell association with tumor vascular endothelial cells and basement membrane coverage are also indicators of vessel stabilization and maturation. Using a systemically delivered fluorescent lectin tracer to mark perfused vessels, we observed a significant increase in tumor perfusion in mice treated with 15C4. Lastly, vessel normalization, through LRG1 antibody blockade, significantly enhanced the efficacy of cisplatin chemotherapy as shown by a slower tumor growth rate and increased tumor cell death compared to monotherapy. These data further corroborate the hypothesis that inhibition of LRG1 improves the delivery, and hence efficacy, of a cytotoxic drug. In conclusion, deletion or inhibition of LRG1 results in an improved vascular configuration and function, and the efficacy of chemotherapy. LRG1 blockade may therefore represent a novel strategy to enhance vessel health and improve the efficacy of cancer therapeutics.
Citation Format: Camilla Pilotti, Marie N O'Connor, David Kallenberg, Laura Dowsett, Jestin George, Stephen E. Moss, John Greenwood. LRG1 blockade normalizes tumor vasculature and improves efficacy of chemotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1477.
Abstract 1480: CCL4 promotes oral squamous cell carcinoma angiogenesis by targeting the angiopoietin-2 signal pathway
Authors: Lien Ming-Yu 
Published: September 2020

Abstract
The inflammatory chemokine (C-C motif) ligand 4 (CCL4) plays an important role in the pathogenesis and progression of cancer, with higher serum CCL4 levels in patients with oral squamous cell carcinoma (OSCC) marking a more advanced stage of disease. OSCC accounts for approximately 95% of oral cancer in Taiwan and this disease has a poor prognosis, with aggressive local invasion and metastasis predisposing to recurrence and additional morbidity. Angiogenesis, a physiological process that enables new blood vessels to form from pre-existing vessels, is involved in the growth, maintenance, and metastasis of most tumors. Angiopoietin-2 (ANGPT2) induces angiogenesis and is a potent endothelial cell-specific mitogen. Until now, it has been unclear as to whether levels of CCL4 expression are a marker of angiogenic activity in patients with OSCC and are correlated with higher levels of ANGPT2 expression. Our investigations have found that treating OSCC cells with CCL4 increases ANGPT2 expression and that CCL4 treatment promotes angiogenesis in a
murine model of OSCC. Thus, CCL4 may be a novel molecular therapeutic target for inhibiting angiogenesis and metastasis in OSCC.
Citation Format: Ming-Yu Lien. CCL4 promotes oral squamous cell carcinoma angiogenesis by targeting the angiopoietin-2 signal pathway [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1480.
Abstract 1489: Exosomes are key players in nicotine-induced angiogenesis
Authors: Singh Shubhangi , Saranyutanon Sirin , Khan Mohammad Aslam , Srivastava Sanjeev Kumar , Singh Seema , Singh Ajay P. 
Published: September 2020

Abstract
Nicotine, an addictive component of cigarette smoke and tobacco, have been shown to promote cancer pathogenesis. Angiogenesis is an important hallmark of cancer that sustains the supply of nutrients and oxygen to the fast proliferating cancer cells through the formation of new blood vessels. The goal of present study was to examine the direct and indirect effect of nicotine on endothelial cell growth, their migration and invasion, and capillary forming ability. We treated the human umbilical vein endothelial cells with two doses of nicotine within physiological range of smokers (200nM and 400nM) and examined the effect on their growth. In parallel, we treated the prostate cancer cells with similar doses of nicotine or vehicle for 8 h and subsequently replaced the media. After next 48 h, conditioned media (CM) was collected from vehicle- (Veh-CM) and nicotine- (Nic-CM) treated prostate cancer cells and used for the treatment of endothelial cells. The growth of endothelial cells increased by ~19 % and ~22 % upon direct treatment of nicotine at 200nM and 400nM doses, respectively. However, a far greater increase was recorded (~128% and ~ 146%, respectively), when endothelial cells were treated with the conditioned media derived from nicotine-treated cancer cells at 200nM and 400nM doses. In next of experiments, we fractionated Veh-CM and Nic-CM (200nM dose) into soluble and exosomal fractions and examined their effect on endothelial cells. We observed that exosomal fraction was more potent (~89%) in inducing endothelial cell growth as compared to the soluble fraction (~46%). In addition, we also observed significant greater increase in motility, invasion and capillary-forming ability of endothelial cells that were pre-treated with Nic-Exo as compared to those treated with Veh-Exo. Studies are currently ongoing to examine the comparative efficacy of nicotine-treated soluble and exosomal factions in potentiating motility, invasion and capillary formation and define the underlying molecular mechanism(s). Our findings thus support a novel pathogenic role of nicotine in cancer where exosomes act as key players in promoting angiogenesis.
Citation Format: Shubhangi Singh, Sirin Saranyutanon, Mohammad Aslam Khan, Sanjeev Kumar Srivastava, Seema Singh, Ajay P. Singh. Exosomes are key players in nicotine-induced angiogenesis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1489.
Abstract 1523: Novel role of TBK1 in breast cancer EMT
Authors: Maan Meenu , Dutta Mainak , Jaiswal Neha , Saavedra Harold , Chellappan Srikumar 
Published: September 2020

Abstract
TANK Binding Kinase 1 (TBK1) regulates interferon signaling and NFκB function by acting as a non-canonical IκB kinase. TBK1 also participates in RalB-mediated inflammatory responses and is essential for the survival of non-small cell lung cancers (NSCLC) driven by oncogenic KRAS. Recent evidence suggests a role for TBK1 as a driver of cancer progression and it is overexpressed in breast, colon, lung and pancreatic cancers. Our published results demonstrate that TBK1 is essential for proper microtubule dynamics and regulation of mitosis. Here we show that knockdown of TBK1 results in a decreased expression of mesenchymal genes involved in EMT and a re-distribution of β-catenin in breast cancer cells. Similarly, expression of matrix metalloproteinases 2 and 9 was altered. This led us to hypothesize that TBK1 acts as an EMT driver in cancer. Inhibition of TBK1 resulted in decreased adherence dependent as well as independent colony formation and stem cell self-renewal, while inducing senescence like phenotypes. We propose that TBK1 mediated EMT is mediated through its interaction with the transcription factor E2F1; the interaction was detected in breast cancer cell lines as well as breast cancer Tissue Micro Arrays. To get a holistic view of the changes that take place in the cell after TBK1 knockdown, we performed a global metabolomics and RNAseq analysis of breast cancer cell lines. An integrated analysis of this data suggested significant alterations in glucose, galactose and nucleotide metabolism upon TBK1 knockdown. Two of the genes that were markedly altered were aldoketoreductase1 B10 (AKR1B10) and thymidine phosphorylase (TP); attention was focused on these genes, as both have been reported to be cancer drivers and their over-expression results in poor prognosis. We observed a downregulation of AKR1B10 and TP in response to the TBK1 knockdown. Experiments are under way to investigate whether the pro-cancer role of TBK1 is dependent on AKR1B10 and TP, especially in breast cancer cells. These results suggest that TBK1 mediated EMT might be a function of the altered expression of mesenchymal proteins and the metabolic profile of the cell brought about by TBK1.
Citation Format: Meenu Maan, Mainak Dutta, Neha Jaiswal, Harold Saavedra, Srikumar Chellappan. Novel role of TBK1 in breast cancer EMT [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1523.