bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3992: Proteomic profiling of breast tumors using a microfluidic quantitative immunohistochemistry platform
Authors: Kim Minseok S. , Kwon Seyong , Lee Jungmin , Lee Eun-Sook , Park Je-Kyun 
Published: February 2011

Abstract
In breast cancers, one of the most troublesome characteristics for cure is the heterogeneity in point of genetic alterations, treatment-response variability, and histopathological features. This characteristic urgently requires personalized tailored therapy, getting out of a simple and widespread prescription. Therefore, the investigation of multiple biomarkers is inevitable for complete recovery. Immunohistochemistry (IHC) has been a major diagnostic method in various malignancies and many studies showing the relationship between immunohistochemical profiles and molecular classification support that IHC might play a significant role in subclassification of breast cancer patients for personalized medicine. However, conventional IHC method is not adequate for examination of multiple biomarkers since it requires consumption of many tissue slices and proportionally increases diagnostic cost which can give a big burden to patients. In addition, subjective decision for biomarker expression is another major problem. Therefore, an innovative multiplexed IHC platform satisfying not only investigation of multiple biomarkers with reducing consumption of tissue slice but also quantification for the biomarkers is the current demand for personalized medicine.
Here, a novel microfluidic multiplexed and quantitative IHC platform was developed for proteomic profiling of breast tumors. The large-scale microfluidic device was designed to have 20 microchannels, and multilayer soft lithography and two-step lithography technologies were used for fabrication. Fluidic resistance was considered for equivalent flow rate of individual reaction channel. Ten biomarkers known as predictive and prognostic indicators in breast cancers, including SMM-HC, CK5, CK14, E-Cadherin, p53, ER, p63, PR, HER2, and Ki-67, were examined on four breast cancer cell lines (MCF-7, SK-BR-3, AU-565, and HCC70) to confirm the massive multiplexed IHC. Results showed that the platform successfully realized the expressions of each biomarker on a cellblock at the same time and proteomic profiling for 10 biomarkers could be quantitatively analyzed from the image analysis. The platform was also applied to a breast cancer tissue sample and the result was corresponded to the results of conventional IHC method. This microfluidic platform is expected to realize precise subtyping of cancer patients by the proteomic profiling even though the size of tumor samples acquired by operation has been getting smaller due to early detection and neoadjuvant therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3992.
Abstract 3995: Differential expression of CD10 immunohistochemistry in stroma and epithelium of prostatic adenocarcinoma
Authors: Fahim Lisa , Lockwood Gina , Fleshner Neil , Sweet Joan 
Published: February 2011

Abstract
Introduction:
CD10 is a transmembrane metallo-endopeptidase involved in cleavage and inactivation of certain peptide hormones important for signal transduction.
CD10 is strongly expressed by normal prostatic luminal epithelial cells. Immunohistochemical (IHC) studies have shown expression of CD10 in prostatic adenocarcinoma. (PCa). Loss of CD10 expression is a common, early event in human prostate cancer (PCa); however, CD10 positive cancer cells frequently appear in lymph node metastasis.
Only a few studies are available on the expression of CD10 in the prostatic stromal and cancer cells. We investigated the IHC expression of CD10 in epithelium and stroma of PCa using TMAs.
Material and Methods:
Prostate tissue from 72 radical prostatectomies was used to construct two TMAs; 35 case of Gleason score (GS) 6(3+3) and 37 cases of high grade (HG) prostate cancer (GS 8, 9, 10) were included. Each case was represented by three, 1mm cores for a total of 105 cores for the GS6 and 111 cores for HG prostate carcinomas. TMAs were stained with H&E and IHC was performed for CD10. Digital images of the TMA slides were scanned using the T3 ScanScope by Aperio Technologies. Epithelial and stromal expression of CD10 was scored as negative or positive.
Results:
CD10 was expressed in epithelium and stroma of PCa. CD10 had heterogeneous expression within individual patients in both epithelium and stroma. This heterogeneity was not significantly different between GS6 and HG prostate cancer.
CD10 was differentially expressed in epithelium of GS6 and HG prostate cancer. HG prostate cancer showed significantly more epithelial staining with CD10 (p=0.001) and less stromal staining with CD10 (p=0.003) compared to GS6.
Conclusion:
GS6 and HG prostate cancer demonstrate a reverse relationship between CD10 expression in the epithelium and the stroma.
Epithelial CD10 expression is an unfavorable risk factor in prostate cancer. Prostate tumors expressing high levels of epithelial CD10 correlate with a poor disease outcome. CD10 may be important in the biology of the tumor stromal microenvironment and its contribution to tumor progression requires further investigation.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3995.
Abstract 4: 5′ - 3′ UTR interactions regulate p53 mRNA translation and provide a target for modulating p53 induction after DNA damage
Authors: Chen Jing , Kastan Michael 
Published: February 2011

Abstract
Induction of p53 protein is a significant determinant of cellular outcome following exposure to DNA damage or other cellular stresses. Though the increase of p53 protein level after DNA damage is largely attributed to an increase in its protein half-life, optimal induction of p53 protein after DNA damage requires RPL26-mediated increases in p53 mRNA translation (Takagi et al., 2005). We report here the existence of a double-strand RNA region containing complementary sequences of the 5′- and 3′- untranslated regions (UTR) of p53 mRNA that is critical for its translational regulation by RPL26. Mutating as few as three bases in either of the two complementary UTR sequences to disrupt this UTR interaction abrogates the ability of RPL26 to bind to p53 mRNA and to stimulate p53 translation. Compensatory mutations that restore the above UTR interaction rescue RPL26 binding as well as translational regulation of p53 by RPL26. Short, single-strand oligonucleotides containing critical sequences from this RNA interaction region inhibit the binding of RPL26 to p53 mRNA and reduce p53 induction and p53-mediated cell death after several different types of DNA damage and cellular stress. These observations demonstrate a novel mechanism of regulating cellular protein levels and provide a reagent (oligonucleotides) that can be used to modulate stress induction of p53 in cells, a potentially clinically useful intervention. These mechanistic insights also set the stage for identifying small molecules other than oligonucleotides which could modulate p53 induction by targeting this mechanism.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4.
Abstract 40: Developmental profile and regulation of the glycolytic enzyme hexokinase 2 and its association with aerobic glycolysis
Authors: Wolf Amparo M. , Agnihotri Sameer , Munoz-Gajadhar Diana M. , Hawkins Cynthia , Guha Abhijit 
Published: February 2011

Abstract
Proliferating embryonic and tumor tissues rely on aerobic glycolysis, or the metabolism of glucose to lactate under oxygenated conditions, to assist in the synthesis of biosynthetic precursors necessary for proliferation. The reliance on aerobic glycolysis may be mediated by the expression of specific metabolic enzymes. Mammalian Hexokinases HK1 and HK2 are 100kDa proteins that phosphorylate glucose to glucose-6-phosphate as the first step of the glycolytic pathway. In normal adult tissues, HK1 is ubiquitously expressed but is particularly prominent in the brain and kidney. HK2 is generally expressed at low levels within adipose tissue and skeletal tissue and negligently in normal brain. In this study, we wished to determine the ontogeny of these isoforms within the developing brain and determine how their expression relates to the extent of aerobic glycolysis in Glioblastoma Multiforme (GBM), a highly aggressive malignant brain tumor. Mining of existing published microarray data found strong expression of HK1 in 1, 2, 4 cell stage but a switch to stronger expression of HK2 in the blastocyst stage, previously reported to rely heavily on aerobic glycolysis. We performed HK2 immunohistochemistry in mice from gestational age E12, E16, post natal 1 month and 2 months. Our results demonstrated an age and cell specific HK1 and HK2 immunoreactivity in embryonic brain tissue with decreased expression of HK2 post-natally. Subsequently, we investigated the expression of HK1 and HK2 isoforms in a panel of GBM cell lines with varying levels of dependence on aerobic glycolysis, as measured by lactate levels and O2 consumption. HK2 but not HK1 expression was higher in GBM cells that had low O2 consumption and high extracellular lactate levels, supporting an association of HK2 with aerobic glycolysis. As HK2 expression is nearly silent in adult brain but expressed in fetal tissue and GBM cells, we hypothesized that DNA methylation/demethylation events may be playing important in its regulation. Adult normal human brain and GBM cell lines that had no HK2 expression were found to be methylated at CpG islands within intron 1 by bisulfite sequencing in contrast to fetal tissue and HK2-expressing GBM cells. The degree of methylation correlated with transcript expression in GBM cell lines. Treatment of U343 and A172 cell lines with 5-azacytidine restored HK2 transcript expression, supporting that HK2 maybe epigenetically regulated. Overall, our results demonstrate that the expression of the HK2 isoform, in contrast to HK1, may be particularly important in tissues relying on aerobic glycolysis for proliferation including embryonic tissue and GBMs.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 40.
Abstract 4002: C-terminal phosphorylation of human GSTP1 by EGFR and site-directed mutation allows JNK reactivation in glioma cells
Authors: Okamura Tatsunori , Ali-Osman Francis 
Published: February 2011

Abstract
In malignant gliomas, the c-jun N-terminal kinase (JNK) pathway regulates tumor growth and resistance to apoptosis, and correlates with histologic grade and epidermal growth factor receptor (EGFR) tyrosine kinase expression. We recently reported that glutathione S-transferase P1 (GSTP1), a major metabolizing and stress response signaling protein that is frequently overexpressed in brain tumors, is a downstream target of EGFR and undergoes EGFR-dependent tyrosine phosphorylation, in vitro and in vivo, resulting in an enhancement of its enzymatic function and increased drug resistance. Although the dimeric GSTP1 exists in a reversible equilibrium with its monomeric form, the monomeric GSTP1 binds to JNK and acts as an endogeneous JNK inhibitor. In our previous study, the EGFR-phosphorylation of GSTP1 shifted the GSTP1 dimer-monomer equilibrium toward the monomeric state and the c-Jun phosphorylation by JNK was enhanced by phosphorylated-GSTP1 and suppressed by unphosphorylated-GSTP1. EGFR activation resulted in dissociation of GSTP1 from the GSTP1-JNK1 complex in a cell-free system and in glioma cells. The C-terminal domain of GSTP1 is reported to be involved in the interaction with JNK. Accordingly, we synthesized the C-terminal domain peptides of human GSTP1 containing the phospho-acceptor Tyr198, unique to hominidae among species, by EGFR (Y198WT) and its phenylalanine-replaced mutant peptide (Y198F). Both the C-terminal domain peptides significantly suppressed the c-Jun phosphorylation by JNK1 in a cell-free system. The binding assay using the synthetic peptides showed that the C-terminal domain peptides of human GSTP1 bound to JNK1 with a lower affinity when Tyr 198 was phosphorylated by EGFR than when it was unphosphorylated. Mutagenesis in human glioma cells revealed that the EGF-induced c-Jun phosphorylation by JNK decreased in the GSTP1-Y198F mutant, whereas increased in the phospho-mimic GSTP1-Y198D mutant, compared with vector control. Together these results indicate that in malignant glioma cells the C-terminal tyrosine phosphorylation of human GSTP1 by EGFR leads to dissociation of GSTP1 from the GSTP1-JNK1 complex. Thus, the GSTP1-JNK1 direct interaction rather than JNK upstream signaling elicits reactivation of JNK. This regulation of JNK signaling function by the EGFR-GSTP1 crosstalk, defines a novel signaling network with a potential to regulate the biology and therapeutic outcome in malignant gliomas. Supported by NIH grants RO1 CA127872, RO1 CA 112519, P50CA108786 and P30-CA14236.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4002.
Abstract 4008: p75NTR is overexpressed in vestibular schwannomas and protects cells from apoptosis due to suppressed c-Jun N-terminal kinase (JNK) activity
Authors: Yue WeiYing , Woodson Erika , Tryggvason Geir , Clark Jason J. , Hansen Marlan R. 
Published: February 2011

Abstract
Vestibular schwannomas (VSs) represent benign Schwann cell (SC) neoplasms arising from the eighth cranial nerve and account for 8-10% of all intracranial neoplasms. Denervation by axotomy leads to increased p75NTR expression, JNK activation, and eventual death of SCs. In contrast to SCs, VS cells survive in the absence of axons and here we investigated the interactions of p75NTR and JNK signaling on the survival response of cultured primary human VS cells. Using qRT-PCR and western blotting, we find that p75NTR is over-expressed in VS cells compared with normal nerve. However, in contrast to normal SCs, treatment of cultured VS cells with pro-NGF, a high affinity ligand for p75NTR, failed to induce apoptosis. We confirmed that VS cells express sortilin, a co-receptor with p75NTR to bind pro-NGF and induce apoptosis. We further show that VS cells display persistent JNK phosphorylation and activity compared with normal SCs. Treatment of VS cultures with the JNK inhibitors, I-JIP or SP600125, results in decreased proliferation, increased apoptosis, and increased accumulation of mitochondrial superoxides. Overexpression of mitochondrial superoxide dismutase prevents apoptosis due to suppressed JNK. These results suggest that persistent JNK contributes to the survival of VS cells, at least in part, by suppressing accumulation of mitochondrial superoxides. Treatment of VS cultures with proNGF protected the cells from apoptosis due to JNK inhibition. This protective effect is specific to proNGF and not seen with other neural derived growth factors such as neuregulin-1 (NRG-1). Indeed treatment with NRG-1 increased apoptosis in cultures with suppressed JNK activity. Further, VS cells lack Trk expression supporting the notion that the protective effect of proNGF is mediated by p75NTR. These results suggest that, in contrast to normal SCs which undergo apoptosis in response to p75NTR and JNK signaling, these signals promote VS cell survival. Such differences may account for the ability of VS cells to proliferate and survive in the absence of axons.
Support: DOD NF050193 and NIDCD R01DC009801
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4008.
Abstract 4009: Identification of Y726 and Y793 as novel phosphorylation sites in human Flt3 and their role in signaling
Authors: Razumovskaya Elena , Sun Jianmin , Rönnstrand Lars 
Published: February 2011

Abstract
Fms-like tyrosine kinase-3 (FLT3), a growth factor receptor expressed in hematopoietic progenitor cells. FLT3 mutations, ITD and D835, are found in approximately 30% of AML patients and correlate with a poor prognosis, thus making receptor a potential therapeutic target. Flt3 mutations cause constitutive activation of the receptor, leading to alteration in signal transduction. The aims of this study are to analyze the kinetics and specificity of FLT3 autophosphorylation in wild-type FLT3 and FLT3 mutants, to identify new phosphorylation sites in FLT3 receptor and to determine related signaling pathways.
Using a panel of “homemade” phosphospecific antibodies directed against potential tyrosine phosphorylation sites in FLT3, we identified three novel phosphorylation sites and studied the kinetics and specificity of phosphorylation in living cells.
Our results show that activation of the wild-type Flt3 receptor is ligand- dependent and responding in a time dependent manner in contrast to FLT3-ITD and D835Y that are constitutive active and ligand-independent. Eight phosphorylated tyrosine residues (pY589, pY591, pY599, pY726, pY768, pY793, pY842 and pY955) were shown to have different phosphorylation characteristics in the wild-type receptor compared to the mutated receptors. Furthermore, we showed that Y726, Y793 and Y842 are novel phosphorylation sites of FLT3 receptor. Further study on stable transfected cells with Y-to-F mutations (726F and 793F), showed that these sites might be important for cell survival, proliferation and transformation. We could also see decreased phosphorylation of Erk1/2 and SHP2 in mutants compare to wild-type FLT3 or FLT3-ITD. Potential binding partners for Y726 and Y793 were predicted by using synthetic phosphopeptides for probing of SH2 domain arrays. The candidate binding partners will be verified as true binding partners using co-immunoprecipitation experiments.
From this study we could observe not only quantitative changes but also qualitative differences in the phosphorylation patterns of wild-type and mutated FLT3 receptors. We have also identified novel phosphorylation sites in FLT3 receptor that can be involved in different signaling pathways. Using this data we can further study the mechanisms of signaling pathways of the Flt3 receptor that are involved in many biological responses and understand the mechanism by which FLT3-ITD and -D835Y functions in pathological conditions.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4009.
Abstract 401: CD5+CD23+ leukemic cell populations in Tcl1 transgenic mice show significantly increased proliferation and Akt phosphorylation
Authors: Efanov Alexey A. , Zanesi Nicola , Maximov Vadim , Nazaryan Natalya , Santanam Urmila , Palamarchuk Alexey , Croce Carlo M. , Pekarsky Yuri 
Published: February 2011

Abstract
B-cell chronic lymphocytic leukemia (B-CLL) is the most common human leukemia. Deregulation of the T-cell leukemia/lymphoma 1 (TCL1) oncogene in mouse B-cells causes a CD5 positive leukemia similar to aggressive human B-CLLs. We recently reported that TCL1 expression in B-CLL is regulated by miR29 and miR181. To determine whether treatment with microRNAs can inhibit
B-CLL in mice by targeting TCL1 we generated TCL1 transgenic mice using a construct containing 3’ and 5’ UTRs of TCL1 under B-cell specific Eμ promoter (Eμ-TCL1fl). At the age of 12-18 months these mice showed B-CLL like disease. Immunophenotyping revealed accumulation of CD5+CD23+B220+ population in spleens and lymph nodes. Our results show that CD5+CD23+ B-cell populations from Eμ-TCL1fl mice actively proliferate and display significantly increased levels of phospho-Akt. Eμ-TCL1fl mice showed immunological abnormalities similar to human B-CLL, including hypoimmunoglobulinemia, abnormal levels of cytokines, and impaired immune response. These findings revealed biochemical and immunological similarities between Tcl1 driven B-CLL in mice and human B-CLL.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 401.
Abstract 4011: CYP3A4 over-expression promotes ER-positive breast cancer cell growth, in part, through Stat3 activation
Authors: Guo Zhijun , Mitra Ranjana , Potter David A. 
Published: February 2011

Abstract
Cytochrome P450 3A4 (CYP3A4) is an important monooxygenase that metabolizes a wide range of xenobitics including many drugs and environmental chemicals, as well as endogenous substances such as steroids. Expression of CYP3A4 has been observed in breast tumors, but its studies in breast cancer cells have mainly focused on carcinogen bioactivation, hormone metabolism and chemotherapy resistance. Whether CYP3A4 has a direct effect on breast cancer progression is unknown. To address these questions, we performed RNAi knock down studies, demonstrating roles for CYP3A4 in proliferation, motility, and clonogenicity of the MCF7 line. CYP3A4 knock down also inhibited phosphorylation of key regulatory proteins such as c-Src and STAT3, implicating its importance in breast cancer progression. To further study the effect and mechanisms of CYP3A4 in breast cancer progression, stable CYP3A4 over-expressing MCF7 cell lines were isolated and characterized. CYP3A4 over-expressing clones exhibited a higher proliferation rate than the vector control clone, consistent with the flow cytometry cell cycle analysis in which CYP3A4 over-expressing clones exhibited a markedly higher S phase component. Apoptosis was not increased due to CYP3A4 over-expression. The CYP3A4 over-expressing clones adhere poorly to plastic and require poly-D-lysine coated tissue culture plate to grow robustly. Adhesion to fibronectin was also reduced as the result of over-expression. In immunofluorescence microscopic study, the highest CYP3A4 over-expressing clone displayed disorganized focal adhesions, potentially explaining the poor adhesion. Consistent with the knock down study, CYP3A4 over-expressing clones exhibited elevated phosphorylation of both c-Src and Stat3 that could be responsible for the observed promotion of proliferation. These results support the model that CYP3A4 is an important signaling protein, linked to the c-Src/Stat3 pathway that influences breast cancer progression.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4011.