bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5281: Role of SHP-2 tyrosine phosphatase in colorectal carcinogenesis
Authors: Sansfacon Jessica Gagne , Coulombe Geneviève , Bourdages Nadia , Rivard Nathalie 
Published: May 2015

Abstract
SHP-2 (Src homology-2 domain-containing phosphatase 2) is a tyrosine phosphatase ubiquitously expressed. Previous work done in SHP-2-/- fibroblasts demonstrated its implication in proliferation and migration as well as in the regulation of many signaling pathways including the Ras-Raf/MEK/ERK, the PI3K/AKT and the Jak/STAT pathways(1). Importantly, gain-of-function mutations were discovered in some colorectal cancers (CRCs)(2). However, the role of SHP-2 in intestinal tumorigenesis has never been investigated.
Methods: SHP-2 expression and catalytic activity were analyzed by Western blot, qPCR and immunohistochemistry in human CRC cells and in human tumours from different stages. SHP-2 expression was also analyzed in polyps (adenomas) from mice bearing a mutation in Apc gene (APCMin/+ mice). To elucidate the role of SHP-2, RNA interference was used to specifically downregulate its expression in intestinal epithelial cells transformed or not by the oncogenic form of KRASG12V (IEC/KRas). Proliferation (cell counting), invasion (through Matrigel in Boyden chamber), anchorage-independent growth (soft agar) and tumor formation in nude mice (xenograft assays) were analyzed.
Results: 1- SHP-2 mRNA and protein levels were significantly increased in human CRC cells and tumors, especially in adenomas, in comparison to healthy adjacent tissues. 2- Interestingly, polyps from APCMin/+ mice also exhibited marked increase in SHP-2 protein expression in comparison to normal adjacent intestinal epithelium. 3- SHP-2 silencing in IEC/KRAS cells markedly reduced their proliferation rate, their growth in soft agar, their capacity to migrate and invade Matrigel and finally, their capacity to form tumor in vivo, in nude mice. 4- Of note, phosphorylated and activated levels of MEK and ERK kinases were significantly decreased following SHP-2 silencing in IECs particularly in those expressing the mutated form of KRAS.
Conclusion: Our data suggest that SHP-2 promotes malignancy of intestinal epithelial cells by sustaining the oncogenic activation of MEK/ERK signalling during intestinal tumorigenesis.
(1)Shi ZQ, Lu W, Feng GS., J Biol Chem. 1998 Feb 27;273(9):4904-8.
(2)Bentires-Alj M. et al., Cancer Res. 2004 Dec 15;64(24):8816-20.
Citation Format: Jessica Gagne Sansfacon, Geneviève Coulombe, Nadia Bourdages, Nathalie Rivard. Role of SHP-2 tyrosine phosphatase in colorectal carcinogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5281. doi:10.1158/1538-7445.AM2014-5281
Abstract 5282: Promotion of skin tumorigenesis by deletion of Ppp6c gene
Authors: Hayashi Katsuhisa , Ogoh Honami , Watanabe Toshio , Tanuma Nobuhiro , Shima Hiroshi 
Published: May 2015

Abstract
Protein phosphatase-6 (PP6) is a member of type-2A protein-Ser/Thr phosphatases, and reportedly regulates several cellular functions including DNA damage responses, NF-kB activation, and mitotic spindle formation. Additionally, some recent papers reported somatic mutations of PPP6c gene, encoding the catalytic subunit of PP6, in about 10 % of melanoma patients, suggesting its potential role as either driver or suppressor of melanoma development.
In this study, we generated mice, in which the PPP6C gene is deleted in keratinocyte-specific manner (PP6-KKO). PP6-KKO mice were predisposed to formation of skin tumor upon DMBA-treatment. Our results demonstrate PPP6c has a tumor-suppressive function at least in skin tumorigenesis.
Citation Format: Katsuhisa Hayashi, Honami Ogoh, Toshio Watanabe, Nobuhiro Tanuma, Hiroshi Shima. Promotion of skin tumorigenesis by deletion of Ppp6c gene. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5282. doi:10.1158/1538-7445.AM2014-5282
Abstract 5285: Protein phosphatases modulate kinase signaling mediated Wnt activation
Authors: Boral Debasish , Nie Daotai 
Published: May 2015

Abstract
Multiple growth factors, signaling through their respective kinase cascades, phosphorylate beta-catenin at specific Serine residues to cause its nuclear sequestration thereby fueling Wnt driven tumor cell proliferation. Our present work identifies a group of phosphatase genes that attenuate such growth factor mediated activation of Wnt signaling in colo-rectal cancer (CRC) cells. This proposed mechanism proposes an additional mode of regulation of Wnt activity in CRC cells.
Using a lentiviral TCF-LEF reporter system, we isolated distinct SW480 (primary) and SW620 (metastatic) cell fractions based on their endogenous level of Wnt activity. Receptor Tyrosine Kinase (RTK) antibody arrays using cell lysates from these cell fractions identified that FGFR3, ALK and RET signaling pathways have a positive correlation with enhanced Wnt activity. Simultaneously qPCR arrays performed on these same cellular fractions identified a total of 10 phosphatase genes whose expression levels are significantly decreased with heightened Wnt activity, out of which 4 genes PHLPP, PHLPPL, PTPRK and PTPN6 are known to regulate the aforementioned RTK pathways.
Of these phosphatase genes, PTPRK has been reported to undergo gene fusion events with RSPO3 (a non-canonical activator of Wnt) in CRC patients. Since the fusion gene product provides no additional Wnt activation that the wild-type RSPO3, we theorized that these fusion events occur either to allow CRC cells to utilize the enhanced promoter activity of the PTPRK gene promoter, or to nullify the Wnt attenuating properties of PTPRK. To test this hypothesis, we have presently cloned and identified PTPRK and RSPO3 gene promoter regions and wish to test their activity in distinct Wnt responsive cell fractions. Our work will allow a better understanding of how phosphatase genes may be involved in controlling the Wnt pathway and how their inactivation may contribute to CRC cell proliferation.
Citation Format: Debasish Boral, Daotai Nie. Protein phosphatases modulate kinase signaling mediated Wnt activation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5285. doi:10.1158/1538-7445.AM2014-5285
Abstract 5289: Role of notch3 in epithelial-to-mesenchymal transition of HCC
Authors: Kunnimalaiyaan Selvi , Gamblin T. Clark , Kunnimalaiyaan Muthusamy 
Published: May 2015

Abstract
Introduction:
The epithelial-mesenchymal transition (EMT) program is often activated in cancer cells during invasion and metastasis. During EMT, expression of transcription factors such as Snail and Slug is induced due to the reduction in E-cadherin expression. In addition, matrix metallo-proteinases (MMPs) which digest the various components of extracellular matrix also induced during EMT. Various signaling pathways have been involved in the regulation of EMT program. We hypothesize that notch signaling pathway is a potential regulator of EMT program in HCC.
Methods:
Lysates from human HCC cell lines, Huh-7, HepG2, and Hep3B, were analyzed for the expression of active notch intracellular domain (NICD) 1-3 proteins by western analysis. To determine the role of NICD3 in cancer invasion and metastasis, siRNA against NICD3 was transfected into HCC cell lines. The effect of depletion of NICD3 in HCC cell lines, epithelial-mesenchymal transition (EMT) markers were measured by western analyses.
Results: Varied level of expression of NICD1-3 was observed in HCC cell lines tested. Transfection of siRNA against NICD3 reduced the level of NICD3 but not the NICD1 protein. Western analysis for the EMT markers expression showed an increase in E-cadherin and concomitant reduction in Snail, Slug, Matrix Metallo Proteinases (MMPs) 2 and 9.
Conclusions:
NICD3 inhibition significantly reduced the expression of markers of metastasis by increasing E-cadherin and suppressing transcription factors Snail, Slug, and MMPs. These observed change in the protein levels are occurred in the absence of NICD3 protein but the presence of NICD1 protein suggesting that NICD3 may play a role in metastasis. Therefore, targeting NICD3 may be potential strategy for treatment of HCC.
Citation Format: Selvi Kunnimalaiyaan, T. Clark Gamblin, Muthusamy Kunnimalaiyaan. Role of notch3 in epithelial-to-mesenchymal transition of HCC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5289. doi:10.1158/1538-7445.AM2014-5289
Abstract 5293: GSK3β phosphorylation at serine9 by RSK2 enhances calcium-induced stress tolerance and cell survival
Authors: Lee Ji-Young , Lee Cheol-Jung , Lee Mee-Hyun , Song Ji Hong , Cho Yong-Yeon 
Published: May 2015

Abstract
Our previous studies demonstrated that RSK2 plays a key role in cell proliferation and transformation induced by tumor promoters such as epidermal growth factor in mouse and human skin cells. However, no direct evidence has been found regarding the relationship between RSK2 and cell survival. In this study, we found that RSK2 interacted and phosphorylated GSK3β at Ser9. Notably, GSK3β phosphorylation at Ser9 was suppressed in RSK2−/− MEFs compared with RSK2+/+ MEFs by stimulation of EGF and calcium ionophore A23187, a cellular calcium stressor. In proliferation, we found that RSK2 deficiency suppressed cell proliferation compared with RSK2+/+ MEFs. In contrast, GSK3β−/− MEFs induced the cell proliferation compared with GSK3β+/+ MEFs. Importantly, RSK2−/− MEFs were induced severe cellular morphology change by A23187 and enhanced G1/G0 and sub-G1 accumulation of the cell cycle phase compared with RSK2+/+ MEFs. The sub-G1 induction in RSK2−/− MEFs by A23187 was correlated with increase of cytochrome c release, caspase-3 cleavage and apoptotic DNA fragmentation compared with RSK2+/+ MEFs. Notably, return back of RSK2 into RSK2−/− MEFs restored A23187-induced morphological change, and decreased apoptosis, apoptotic DNA fragmentation and caspase-3 induction compared with RSK2−/− /mock MEFs. Taken together, our results demonstrated that RSK2 plays an important role in stress-tolerance and cell survival, resulting in cell proliferation and cancer development.
Citation Format: Ji-Young Lee, Cheol-Jung Lee, Mee-Hyun Lee, Ji Hong Song, Yong-Yeon Cho. GSK3β phosphorylation at serine9 by RSK2 enhances calcium-induced stress tolerance and cell survival. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5293. doi:10.1158/1538-7445.AM2014-5293
Abstract 5300: Acetylation of alteration/deficiency in activation 3 (Ada3) is essential for its stability and function
Authors: Srivastava Shashank , Mohibi Shakur , Band Hamid , Band Vimla 
Published: May 2015

Abstract
In eukaryotes histone acetylation is one of the major epigenetic modifications by which gene expression is regulated. Acetylation of histones is mediated by histone acetyl transferases (HATs) that exist as multisubunit complex. The alteration/deficiency in activation 3 (Ada3) protein was discovered as an essential component of various HAT complexes and later on we identified Ada3 as a novel human papilloma virus E6 oncoprotein binding protein. We have previously demonstrated that Ada3 regulates the function of p53 by mediating its acetylation and recent findings from our laboratory show that Ada3 is a novel cell cycle regulator and is crucial for maintaining the genomic stability.
Although Ada3 is an essential mediator of histone acetylation it does not have HAT activity per se and how Ada3 performs its function remains unresolved. Our recent studies demonstrate that Ada3 itself is acetylated by GCN5 and p300 both in in-vitro and in-vivo assays. Significantly, inhibition of acetylation by garcinol dramatically decreases Ada3 protein levels, suggesting that acetylation stabilizes Ada3 protein. Current studies are directed towards identification of potential acetylation sites in Ada3 through mutational analyses. Notably, inhibition of class III but not class I and II histone deacetylases (HDACs) enhanced Ada3 acetylation. Further investigation reveals that Ada3 specifically interacts with SIRT1, a member of class III HDAC, indicating that SIRT1 might deacetylate Ada3. Overall our findings suggest that Ada3 exists in dynamic balance of acetylation and deacetylation, and its cell cycle regulatory function may depend on Ada3 acetylation and deacetylation.
Citation Format: Shashank Srivastava, Shakur Mohibi, Hamid Band, Vimla Band. Acetylation of alteration/deficiency in activation 3 (Ada3) is essential for its stability and function. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5300. doi:10.1158/1538-7445.AM2014-5300
Abstract 531: MiR-199a-5p is a biomarker for and regulator of epithelial-mesenchymal transition in triple-negative breast cancer patients
Authors: Shin Vivian Y. , Siu Man T. , Ho John C. , Kwong Ava 
Published: May 2015

Abstract
Objectives: Triple-negative breast cancer (TNBC) accounts for 15-20% of all breast cancer, and is characterized by the absence of estrogen receptor, progesterone receptor and human epidermal growth factor receptor. The lack of targeted therapy is a major obstacle for treating against this aggressive subtype, hence the search of specific biomarkers may use as potential diagnostic marker and perhaps a therapeutic target for TNBC. Methods: Plasma samples from patients with TNBC and non-TNBC were recruited for miRNA profiling. By using miRCURY LNA array containing 730 miRNAs, we compared the differential miRNA expressions in plasma of patient with TNBC (n=5) and non-TNBC (n=5), as well as healthy controls (n=5). Selected miRNAs were further validated in an independent cohort (n=250) using real-time RT-PCR. Functional study of miRNA was also carried out in the cell lines. Results: Microarray data revealed miR-16, miR-21 and miR-199a-5p have differentially expression between TNBC and non-TNBC. We found that miR-16, miR-21 and miR-199a-5p were underexpressed in TNBC cases when compared with healthy controls. Moreover, post-operative expression levels of miR-16, miR-21 and miR-199a-5p were significantly increased than that of pre-operative plasma of TNBC. We then stratified the patients by TNM stage and associate with miRNA expression level. Plasma miR-199a-5p expression in TNBC patients had significant difference when compared with healthy controls (stage I, p&lt;0.002; stage II, p&lt;0.001; stage III, p&lt;0.011). Transfection of miR-199a-5p mimic significantly reduced cell proliferation and restored the epithelial-mesenchymal transition (EMT) markers (E-cadherin, ZEB1 and TWIST2). Conclusions: Our data implicate that miR-199a-5p is a potential marker for discriminating TNBC cases and non-TNBC cases. The association of miR-199a-5p with EMT markers uncovers an important pathway in metastatic breast cancer. This study provides insights for the potential use of miRNA as diagnostic marker and targeted therapy for the treatment of TNBC.
Citation Format: Vivian Y. Shin, Man T. Siu, John C. Ho, Ava Kwong. MiR-199a-5p is a biomarker for and regulator of epithelial-mesenchymal transition in triple-negative breast cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 531. doi:10.1158/1538-7445.AM2014-531
Abstract 5312: Quantitative proteomic analysis of cell-surface membrane proteins: Biomarker discovery in esophageal squamous cancer
Authors: Harada Emi , Serada Satoshi , Takahashi Tsuyoshi , Fujimoto Minoru , Naka Tetsuji 
Published: May 2015

Abstract
Background. Esophageal squamous cell cancer (ESCC) is associated with a poor prognosis and high recurrence rate. Identification of proteins that are useful for prognostic assessment and therapeutic targets in ESCC is required. Quantitative proteomic analysis provides a powerful approach in screening for alterations in protein levels. An analysis of the differential expression of cell-surface membrane proteins would be useful for identification of proteins associated with cell proliferation, cell migration, drug resistance and therapeutic target for antibody-based therapy.
Purpose of study. To identify cell-surface membrane proteins overexpressed in ESCC compared with normal esophageal epithelial cells, which may represent novel biomarkers of disease and therapeutic targets.
Procedures. We performed a biotinylation-based approach for cell-surface membrane enrichment combined with iTRAQ (isobaric tags for relative and absolute quantitation) technology using nano-LC-MS/MS analysis, to identify cell surface proteins overexpressed in ESCC cell lines compared with control normal esophageal epithelial cell line.
Results. By combining biotinylation of cell-surface membrane proteins and iTRAQ analysis, 1635 proteins were identified, which contained 402 cell surface membrane proteins. We identified overexpression of 12 membrane proteins, whose expression increased more than twofold in at least four of six ESCC cell lines compared with a normal esophageal epithelial cell line (HEEpic). By this proteomic approach, we identified several tumor antigens which included antigens previously associated with ESCC and antigens not previously reported in this disease. Furthermore, we also successfully identified novel antigen associated with proliferation of ESCC. Cell surface expression of this antigen on ESCC was confirmed by FACS analysis and immunohistochemistry. By transfection of siRNA specific for this novel antigen, significant growth suppression was shown by WST8 assay and induction of apoptosis was detected by caspase-3 assay.
Conclusions. By using a high throughput cell-surface membrane proteomic analysis combining biotinylation of cell-surface membrane protein and iTRAQ technology, several ESCC candidate antigens were identified. Further studies on the physiological role of novel tumor antigens overexpressed in ESCC and their application as biomarkers/therapeutic targets are underway.
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Note: This abstract was not presented at the meeting.
Citation Format: Emi Harada, Satoshi Serada, Tsuyoshi Takahashi, Minoru Fujimoto, Tetsuji Naka. Quantitative proteomic analysis of cell-surface membrane proteins: Biomarker discovery in esophageal squamous cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5312. doi:10.1158/1538-7445.AM2014-5312
Abstract 5318: LC-MS/MS quantification of proteins CD47 and SIRPA in cell subtypes selected by flow cytometry from blood
Authors: Fernandez-Metzler Carmen , Capocasale Renold 
Published: May 2015

Abstract
Flow cytometry and mass spectrometry are widely used analytical tools in cancer research. Flow cytometry is known for cell counting, cell sorting and cancer biomarker discovery. Mass spectrometry is unparalleled in its ability to selectively detect and quantify target analytes at the molecular level. The combination of flow cytometry to isolate specific cell subtypes from biological fluids such as blood and mass spectrometry to quantify proteins contained in the selected cells provides a new approach to studying protein expression in specific cell subtypes. We have used this combination to investigate the normal levels of CD47 and SIRPA proteins in CB8+ T-cells, CD4+ T-cells, CD14+ monocytes, CD33+ myeloid cells and CD56+ NK cells isolated from blood specimens as a first step toward better understanding of how the CD47/SIRPA protein levels in these cells are affected by cancer and cancer treatment.
Citation Format: Carmen Fernandez-Metzler, Renold Capocasale. LC-MS/MS quantification of proteins CD47 and SIRPA in cell subtypes selected by flow cytometry from blood. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5318. doi:10.1158/1538-7445.AM2014-5318
Abstract 5320: Novel computational methods reveal subtle clonal mosaicism
Authors: Scheet Paul , Xia Rui , Lucas F. Anthony San , Hahn Christina , Fowler Jerry , Vattathil Selina 
Published: May 2015

Abstract
Recent genome-wide surveys have revealed age-related genetic mosaicism and established a link between clonal mosaicism and risk of hematological and solid-tumor cancers (Laurie et al, 2012, Nat Genet, 44:642; Jacobs et al, 2012, Nat Genet, 44:651; Forsberg et al, 2012, AJHG, 90:217). Existing methods to identify chromosomal aberrations leading to mosaic allelic imbalance (AI) rely on the detection of increased variance of the estimated within-sample allele frequencies at heterozygous loci, ignoring the dependence among allele frequencies in regions of an AI-inducing event, such as a deletion or copy-neutral loss of heterozygosity (LOH), expected from the loss or gain of a chunk of an inherited chromosome. As a result, they break down when the proportion of DNA from an aberrant source is below 10-15%. Here we present a suite of new methods (hapLOH, J-LOH, hapLOHseq) that harness the important information contained in the germline haplotypes to help distinguish signal from noise, accommodate multiple samples of different tumor purities (to deconvolute aberration type and tumor purity) and infer co-occurrence of aberrations in heterogeneous mixtures. For example, our initial method, hapLOH (Vattathil & Scheet, 2013, Gen Res 23:152), can identify 10 Mb regions with just 4% of the signal coming from the tumor genome in data simulated from a series of experiments from a 317K SNP microarray. Application of hapLOH to data analyzed by Laurie et al (2012) indicates that we have substantially greater sensitivity to detect clonal mosaicism, particularly events harbored in a small proportion of cells (&lt; 5-10%). We identify more than twice as many mosaic events, demonstrating the potential role for our methods to aid in profiling cancer risk. Our suite of tools is designed for both microarray and next-generation sequencing data and is available at scheet.org.
Citation Format: Paul Scheet, Rui Xia, F. Anthony San Lucas, Christina Hahn, Jerry Fowler, Selina Vattathil. Novel computational methods reveal subtle clonal mosaicism. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5320. doi:10.1158/1538-7445.AM2014-5320