bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1388A: Assessment of the anti-angiogenic effect of VEGFR2 siRNA in HUVEC using the Lonza 4D-Nucleofecto system
Authors: Kokatam Srinivasan , Tiwari Kanchan , Schroeder Jenny , Toell Andrea , Hussain Lubna , Kapoor Preeti 
Published: August 2015

Abstract
Angiogenesis is a hallmark of most cancers, and is thus an attractive target for the treatment of cancer. One of the easiest screening and target validation strategies for anti-angiogenic target identification involves knocking down targets in Human Umbilical Vein Endothelial Cells (HUVEC) and assessing subsequent effects on tube formation in Corning's Matrigel product.
The usage of small interfering RNA (siRNA) is one of the strategies to knock-down RNA, and thereby protein expression within cells. siRNA can be delivered within cells using either chemical transfection or electroporation-based strategies such as the one offered by the Lonza Nucleofector™ Technology.
In the current study we used the Lonza 4D-Nucleofector™ X-Unit to transfect single-donor Clonetics™ HUVEC cultured in EGM™-2 Endothelial Cell Growth Media. Cells were transfected with siRNA directed against Vascular Endothelial Growth Factor Receptor 2 (VEGFR2). Transfection conditions were fine-tuned using pmaxGFP™ vector, and it was shown that the Nucleofection™ process had no deleterious effect on the tube formation potential of HUVEC on Corning's Matrigel product.
Efficient knock-down of VEGFR2 protein levels was demonstrated after the transfection of VEGFR2-siRNA. Best knock-down efficiency was observed 24 hours after transfection. VEGFR2-siRNA transfected cells demonstrated significant inhibition of tube formation on Corning's Growth Factor Reduced Matrigel product in comparison to control samples.
Since the efficient knock-down of the VEGFR2 protein in Clonetics™ HUVEC using the Nucleofector™ Technology can be demonstrated at time points as early as 24 hours after transfection, this is an efficient approach for target identification, validation and screening of siRNA based anti-angiogenesis therapeutics for cancer treatment.
Citation Format: Srinivasan Kokatam, Kanchan Tiwari, Jenny Schroeder, Andrea Toell, Lubna Hussain, Preeti Kapoor. Assessment of the anti-angiogenic effect of VEGFR2 siRNA in HUVEC using the Lonza 4D-Nucleofecto system. [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 1388A. doi:10.1158/1538-7445.AM2015-1388A
Abstract 139: Activating ERBB4 mutations in non-small cell lung cancer
Authors: Kurppa Kari J. , Denessiouk Konstantin , Johnson Mark S. , Elenius Klaus 
Published: August 2015

Abstract
Genes encoding the ErbB receptor tyrosine kinases (EGFR/ERBB1, ERBB2, ERBB3, and ERBB4) are key regulators of cellular proliferation, survival, and differentiation, and thus represent potent proto-oncogenes. In particular, mutations or copy number variations of EGFR or ERBB2 are present in human malignancies and serve as predictive markers for targeted therapies. Recent efforts to comprehensively characterize the mutational landscape of human cancers have identified frequent somatic mutations in ERBB4 in various cancer types, such as non-small cell lung cancer (NSCLC), melanoma, and colorectal cancer. However, the significance of mutated ERBB4 in cancer remains elusive.
Here, we have functionally characterized nine ERBB4 mutations previously identified in lung adenocarcinoma. Four out of the nine mutations, Y285C, D595V, D931Y and K935I, were found to be activating, increasing both basal and ligand-induced ErbB4 phosphorylation. According to structural analysis, the four activating mutations were located at critical positions at the dimerization interfaces of the ErbB4 extracellular (Y285C, D595V) and kinase (D931Y and K935I) domains. Consistently, the mutations enhanced ErbB4 dimerization and increased the trans activation in ErbB4 homodimers and ErbB4/ErbB2 heterodimers. The expression of the activating ERBB4 mutants promoted survival of NIH 3T3 cells in the absence of serum. Interestingly, serum starvation of NIH 3T3 cells expressing the ERBB4 mutants only moderately increased the phosphorylation of canonical ErbB signaling pathway effectors Erk1/2 and Akt as compared to wild-type ERBB4. In contrast, the mutations clearly enhanced the proteolytic release of signaling-competent ErbB4 intracellular domain.
These results suggest the presence of activating, oncogenic mutations of ERBB4 in non-small cell lung cancer.
Citation Format: Kari J. Kurppa, Konstantin Denessiouk, Mark S. Johnson, Klaus Elenius. Activating ERBB4 mutations in non-small cell lung 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 139. doi:10.1158/1538-7445.AM2015-139
Abstract 1393: CD44v3-positive (CD44v3+) and CD24-negative (CD24−) cells possess cancer stem cell (CSC)-like properties in a human oral squamous cell carcinoma cell line
Authors: Todoroki Keita , Ogasawara Sachiko , Akiba Jun , Nakayama Masamichi , Naito Yoshiki , Kusukawa Jingo , Yano Hirohisa 
Published: August 2015

Abstract
Background:
CSCs are defined as minor cell population which have higher tumor-initiating ability, self-renewal ability and multilineage potential. Although CSCs in oral squamous cell carcinoma (OSCC) were identified by several surface antigen (e.g., CD44, CD133) or ALDEFLOUR assay, it is still elusive which are universal markers for CSC in OSCC. The aim of this study is to clarify potential CSC markers for a OSCC cell line using a combination of cell surface markers, CD44v3 and CD24.
Materials and Methods: A OSCC cell line, SAS, was used in the experiment. CD44v3+/CD24− cell fraction was sorted from SAS cells with a FACSAria II and compared with the other three fractions (CD44v3+/CD24+, CD44v3− /CD24−, CD44v3−/CD24+ cell fraction) in cell proliferation, drug resistance and sphere forming ability. mRNA expression levels of various genes, such as stemness genes, anti-apoptosis genes and hypoxia-related genes, were compared among the fractions using quantitative real-time reverse transcriptase-polymerase chain reaction.
Results:
The proportion of CD44v3+/CD24−, CD44v3+/CD24+, CD44v3−/CD24−, CD44v3−/CD24+ cells was 34.9%, 39.8%, 14.2% and 11.0%, respectively. CD44v3+/CD24− cells showed a higher sphere forming ability, and drug resistance for CDDP, 5-FU and Cetuximab and expressed higher mRNA levels of CSC property-related genes (ABCG2, Oct-4, Nanog) and a hypoxia-related gene (HIF-1α) than the other cell fractions. There were no significant differences in cell proliferation among the fractions.
Conclusion:
The results suggest that CD44v3+/CD24− cell fractions in SAS possess CSC-like properties. The investigation of tumorigenic capacity in NOD/SCID mice is now under way.
Citation Format: Keita Todoroki, Sachiko Ogasawara, Jun Akiba, Masamichi Nakayama, Yoshiki Naito, Jingo Kusukawa, Hirohisa Yano. CD44v3-positive (CD44v3+) and CD24-negative (CD24−) cells possess cancer stem cell (CSC)-like properties in a human oral squamous cell carcinoma cell line. [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 1393. doi:10.1158/1538-7445.AM2015-1393
Abstract 14: Nucleolin phosphorylation by CK2 is important for its role in regulating apoptosis
Authors: Akinwunmi Esther , Xiao Shu , Guo Yuqi , Li Xin , Saxena Anjana D. 
Published: August 2015

Abstract
Nucleolar phosphoprotein nucleolin integrates critical cellular processes including gene expression, ribosome biogenesis, proliferation and the cellular response to stress. Elevated levels of nucleolin are found in a variety of tumors. Nucleolin has distinct mechanisms in p53 regulation to influence cell survival: In “normal unstressed” cells (with hyper-proliferative signals but no obvious DNA damage), nucleolin stabilizes p53 and induces apoptosis. Under DNA-damage however, nucleolin can cause translational repression. The translational repression is overcome upon DNA damage in part, possibly due to nucleolin translocation to the nucleoplasm, altered interactions with ribosomal protein, RPL26 and/or p53-antagonist, HDM2. In contrast, nucleolin binding to AU rich elements (AREs) of anti-apoptotic BCL2-mRNA stabilizes BCL2, leading to cell survival. Importantly, increased nucleolin phosphorylation by the interphase kinase CK2 is finely regulated in exponentially dividing cells; however, role of nucleolin phosphorylation remains largely unexplored in cell survival. We recently demonstrated that phosphorylation deficient nucleolin mutant confers dominant negative effect on cell proliferation. In this study, we dissect the role of nucleolin phosphorylation in regulating p53-signaling pathway. Our pathway specific qRT-PCR-array analyses indicate that expression of phosphorylation-deficient nucleolin mutant resulted in an increased gene expression in the apoptosis, DNA damage and repair pathways that are downstream to p53 activation. We further elucidate that targeting nucleolin phosphorylation by CK2 initiates an increased expression of BH3-only proteins: p53-transcriptional targets e.g. BID and PUMA. Interestingly, an increased expression of BIM, a p53-independent target that is required during DNA damage induced cell death is evident with NCL expression regardless of its phosphorylation status. This is the first step in understanding nucleolin-phosphorylation mediated regulation of gene expression in the p53-pathway to control cell proliferation. This information will be useful in identifying new targets that can be either directly (via RNA binding) or indirectly (through other proteins or gene expression) regulated by nucleolin phosphorylation.
Citation Format: Esther Akinwunmi, Shu Xiao, Yuqi Guo, Xin Li, Anjana D. Saxena. Nucleolin phosphorylation by CK2 is important for its role in regulating apoptosis. [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 14. doi:10.1158/1538-7445.AM2015-14
Abstract 1401: Low proteasome activity and cancer stemness in colorectal cancer
Authors: Uemura Mamoru , Munakata Koji , Nishimura Junichi , Hata Taishi , Takemasa Ichiro , Mizushima Tsunekazu , Yamamoto Hirofumi , Doki Yuichiro , Mori Masaki 
Published: August 2015

Abstract
Background and aims:
Emerging evidence indicates that existence of cancer stem-like cells (CSCs) is one of the main reasons of treatment resistance. Recently, several reports have indicated that some CSCs would be characterized by low proteasome activity.
Material and methods:
We engineered human colorectal cancer (CRC) cells to express a green fluorescent molecule fused to the degron of ornithine decarboxylase (Gdeg) from a retroviral vector; the fluorescent Gdeg supposed to be accumulated in CSCs as a result of low activity of the 26S proteasome. The feature of Gdeghigh cells (low proteasome activity cells; LPACs) were analyzed by sphere formation assay, surface marker analysis, reactive oxygen species (ROS) analysis and assessment of chemotherapy and radiation therapy sensitivity. In addition, we explored the mechanism of treatment resistance of LPACs by using microarray analysis and clinical samples.
Results:
LPACs of CRC cells had significant radioresistance and chemoresistance (5-FU and oxaliplatin). Cancer stemness of LPACs was confirmed by CSC marker, sphere formation assay, ROS activity analysis, cell cycle assay, and in vivo tumorigenicity examination. Microarray analysis identified EID3 (EP300 Interacting Inhibitor of Differentiation 3) as a key factor related to treatment resistance of LPACs. Functional analyses of EID3 and analysis using clinical samples confirmed the role of EID3 in LPACs.
Conclusions:
CRC with low proteasome activity had character of treatment resistance and capacity of stemness. EID3 was a key factor related to treatment resistance of LPACs, and might be a novel therapeutic target.
Citation Format: Mamoru Uemura, Koji Munakata, Junichi Nishimura, Taishi Hata, Ichiro Takemasa, Tsunekazu Mizushima, Hirofumi Yamamoto, Yuichiro Doki, Masaki Mori. Low proteasome activity and cancer stemness in colorectal 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 1401. doi:10.1158/1538-7445.AM2015-1401
Abstract 1410: Evaluation of the immunogenicity of ALDHhigh human head and neck squamous cell carcinoma cancer stem cells in vitro
Authors: Prince Mark E.P. , Zhou Li , Xia Yang , Tao Huimin , Moyer Jeffrey S. , Lu Lin , Owen John , Chang Alfred E. , Xia Jianchuan , Wolf Gregory , Wicha Max S. , Ren Xiubao , Bao Yangyi , Huang Shiang , Li Qiao 
Published: August 2015

Abstract
Using mouse models we reported that dendritic cells (DC) pulsed with cancer stem cells (CSC) enriched by virtue of their expression of the CSC marker aldehyde dehydrogenase (ALDH) significantly reduced development of pulmonary metastases and prolonged survival. In this recent study, we established the concept that the antigenicity/immunogenicity of ALDHhigh human head and neck squamous cell carcinoma (HNSCC) cancer stem cells is distinct from that of ALDHlow non-CSCs. To this end, we generated CSC-loaded DCs to sensitize autologous peripheral blood lymphocytes to react with CSCs using human HNSCC samples in vitro. From peripheral blood and paired tumor tissues collected from 9 patients with HNSCC, we obtained PBMCs and established 4 HNSCC tumor cell lines. DCs generated from the PBMC and pulsed with the lysate of ALDHhigh cells isolated from cultured HNSCC cells (CSC-DC) could sensitize autologous T, B lymphocytes in vitro, which was evident by cytokine production, CTL activity, and antibody secretion of these primed T, B cells in response to ALDHhigh CSCs. In contrast, DCs pulsed with lysate of ALDHlow cells from the same HNSCC patient (ALDHlow-DC) resulted in limited sensitization/priming of autologous T, B lymphocytes to produce IFNγ, lyse CSCs, and secrete IgM and IgG in response to ALDHhigh CSCs. These results demonstrated significant differences in the antigenicity/immunogenicity between ALDHhigh CSCs vs. ALDHlow cells isolated from the tumor specimen of patients with HNSCC, which indicates the existence of unique CSC antigens in the ALDHhigh population. In addition, this study demonstrates that it is feasible to generate DCs from the PBMCs and isolate ALDHhigh CSCs from cultured tumor cells of the patients with HNSCC to prepare CSC-DC vaccines for clinical application.
Citation Format: Mark E.P. Prince, Li Zhou, Yang Xia, Huimin Tao, Jeffrey S. Moyer, Lin Lu, John Owen, Alfred E. Chang, Jianchuan Xia, Gregory Wolf, Max S. Wicha, Xiubao Ren, Yangyi Bao, Shiang Huang, Qiao Li. Evaluation of the immunogenicity of ALDHhigh human head and neck squamous cell carcinoma cancer stem cells in vitro. [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 1410. doi:10.1158/1538-7445.AM2015-1410
Abstract 1413: Significance of KIFC1 in spheroid formation of gastric cancer cells
Authors: Oue Naohide , Mukai Shoichiro , Pham T.B. Trang , Sentani Kazuhiro , Ito Reiko , Yasui Wataru 
Published: August 2015

Abstract
Spheroid colony formation is a typically method to identify cancer stem cells (CSC). The growth of spherical colonies is considered indicative of self-renewal ability, and is consistent with a CSC phenotype. However, mechanism of spheroid formation remains unclear in gastric cancer (GC). In the present study, we analyzed spheroid formation associated genes in GC. Comprehensive gene expression profile of MKN-45 and MKN-74 GC cell lines that were grown as monolayers or spheroids was obtained by Affymetrix GeneChip, and we found several kinds of genes whose expression was up-regulated or down-regulated in cells that were grown as spheroids. Among these, 10 kinds of kinesin genes were up-regulated in both MKN-45 and MKN-74 cells that were grown as spheroids. Kinesins are categorized into 14 subfamilies and classified as mitotic kinesins, which are involved in cell division, and non-mitotic kinesins, which are principally involved in intracellular transport. In spheroids, mitotic kinesins were up-regulated. In the present study, we focused on KIFC1, because antibody against KIFC1 is available. qRT-PCR analysis revealed that up-regulation of KIFC1 in both MKN-45 and MKN-74 cells that were grown as spheroids. Immunohistochemical analysis of KIFC1 was performed in 20 GC cases. Expression of KIFC1 was up-regulated in GC tissues in comparison with corresponding non-neoplastic mucosa. These results suggest that KIFC1 plays an important role in gastric carcinogenesis and may be a therapeutic target of GC.
Citation Format: Naohide Oue, Shoichiro Mukai, Trang Pham T.B., Kazuhiro Sentani, Reiko Ito, Wataru Yasui. Significance of KIFC1 in spheroid formation of gastric 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 1413. doi:10.1158/1538-7445.AM2015-1413
Abstract 1416: Granulocyte-colony stimulating factor (G-CSF) enhances stemness by inducing myeloid-derived suppressor cell (MDSC) in cervical cancer
Authors: Kuroda Hiromasa 
Published: August 2015

Abstract
Objectives: Granulocyte-colony stimulating factor (G-CSF) is known to induce the myeloid-derived suppressor cell (MDSC) to stimulate the progression of uterine cervical cancer. The aim of the current study is to investigate the role of MDSC in the induction of cancer stem cells in uterine cervical cancer.
Methods: We first established cervical cancer cell lines stably transfected with G-CSF expression vector (ME180-GCSF) or control vector (ME180-control). Then, using BALB/c nude mice subcutaneously inoculated with ME180-G-CSF or ME180-control cells, the effect of G-CSF on the induction of MDSCs (CD11b+Gr1+ cells) and cancer stem cells (ALDH-high cells) were assessed by flow cytometry. Using MDSCs isolated from the spleens of ME180-GCSF-bearing mice, we next investigated whether the MDSCs induce cancer stem cells in vitro. Finally, we analyzed the association between the G-CSF expression and ALDH1A1 expression in human cervical cancer specimens by immunohistochemistry.
Results: 1) G-CSF induces MDSCs from mice bone marrow. 2) ME180-GCSF-derived tumors contain greater number of MDSCs and ALDH-high cells. 3) Co-incubation of ME180 with MDSCs cells increased the number of ALDH-high cells. 4) A positive correlation was observed between the tumor G-CSF expression and ALDH1A1 expression in human cervical cancers.
Conclusions: G-CSF-induced MDSCs increase the number of cancer stem cells in uterine cervical cancer.
Citation Format: Hiromasa Kuroda. Granulocyte-colony stimulating factor (G-CSF) enhances stemness by inducing myeloid-derived suppressor cell (MDSC) in cervical 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 1416. doi:10.1158/1538-7445.AM2015-1416
Abstract 1418: Screening and identification of key surface markers of mesenchymal stem cell osteogenic differentiation and osteosarcoma development
Authors: Ren Tingting , Piperdi Sajida , Park Amy Y. , Gorlick Richard 
Published: August 2015

Abstract
Introduction: Osteosarcoma (OS) is the most common primary malignant tumor of bone in children and young adults. Knowledge about its cell of origin is limited, making new treatments difficult to develop. Our aim is to screen and identify the key surface markers of mesenchymal stem cell (MSC) to osteogenic differentiation and OS malignant transformation.
Methods: A list of differentially expressed genes with a statistically significant p-value &lt;0.05 was obtained by microarray analysis from MSCs and osteoblasts (OB). MSCs were differentiated into osteoblasts, adipocytes and chondrocytes and confirmed by alizarin red staining, oil red O staining and collagen II immunohistchemistry, respectively. Western blots were then performed to identify the expression of selected surface markers during the MSC differentiation and compared to those of OS cells.
Results: Nine genes were obtained with a p-value &lt;0.02, which are differentially expressed between MSC and OB. Four of them were correlated with cell adhesion and metastasis. The expression profile of CD49b was distinct, where it was down regulated only during MSC osteogenic differentiation and over expressed in OS cell lines.
Conclusions: Cell adhesion may have an important role during MSC differentiation. CD49b is a unique marker during MSC osteogenic differentiation and may play a significant role in osteoasarcoma malignant transformation. In the future we plan to determine the function and mechanism of CD49b during MSCs osteogenic differentiation and osteosarcoma malignant transformation in vitro. We will define the clinical significance of CD49b in osteosarcoma through immunohistochemistry of patient samples and functional analyses.
Citation Format: Tingting Ren, Sajida Piperdi, Amy Y. Park, Richard Gorlick. Screening and identification of key surface markers of mesenchymal stem cell osteogenic differentiation and osteosarcoma development. [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 1418. doi:10.1158/1538-7445.AM2015-1418