bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1088: Characterizing the effects of alcohol on the ER-alpha status of HER2 breast cancer
Authors: Wong Amy W. , Smith Sarah M. , Nunez Nomeli P.
Published: June 2012

Alcohol consumption is a major risk factor for breast cancer. Approximately 400,000 cases of cancer in 2002 were attributable to alcohol consumption worldwide. In women, breast cancer accounts for 60% of these alcohol-related cancers. Over 25% of all breast cancer cases are HER2 (ErbB2) positive and have poor prognosis. Unfortunately, the mechanism by which alcohol promotes breast cancer is unclear.

Our research focuses on characterizing the effects of alcohol on HER2 positive breast cancer. More specifically, we determine if and how alcohol affects the ER-alpha status of ErbB2-expressing tumors in the presence and absence of ovarian hormones using the MMTV-neu mouse model of ErbB2-induced breast cancer. We show alcohol consumption increases tumor incidence as well as tumor proliferation as demonstrated by Ki67 immunohistochemical staining. We measure systemic estrogen levels and ER-alpha expression in the mammary tumors developed from alcohol and water consuming mice to determine if their levels are associated with the higher tumor incidence in alcohol consuming mice. We show that both systemic estrogen and ER-alpha expression levels are elevated in alcohol consuming mice. Our data suggest that alcohol increases tumor development in HER2 transgenic mice via the estrogen pathway. In fact, the effects of alcohol on tumor incidence were blocked with ovariectomy, suggesting that indeed the effects of alcohol on HER2 tumors are mediated via the estrogen pathway.

Our results suggest that the effects of alcohol on HER2 positive breast cancers can be prevented by targeting or blocking the estrogen pathway. For this purpose, we will determine in future studies if the effects of alcohol can be prevented with tamoxifen, a selective ER modulator (SERM), and letrozole, an aromatase inhibitor, both of which blocks the estrogen pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1088. doi:10.1158/1538-7445.AM2011-1088
Abstract 1092: Insulin-like growth factors modulate chemosensitivity to cisplatin and 5-fluorouracil in human esophageal cell lines
Authors: Zhao Ronghua , Casson Alan G.
Published: June 2012

Resistance to chemotherapy remains a major obstacle to survival of patients with esophageal adenocarcinoma (EADC). The aim of this study was to evaluate whether insulin-like growth factors (IGFs), recently implicated in the molecular pathogenesis of EADC, modulate chemosensitivity to Cisplatin (CDDP) and 5-Fluoruracil (5-Fu), two agents widely used in current clinical practice.

Expression of IGF1, its receptor (IGF1R), and IGF2 (mRNA and protein) were studied using quantitative (real-time) PCR and Western blot in human esophageal cell lines: Het1A (derived from immortalized normal esophageal epithelium), OE33 and JHEsoAd1 (each derived from a primary EADC). For each cell line, the effect of various concentrations of IGFs, CDDP and 5-Fu, alone and in combination, on cell proliferation and clonogenicity were evaluated by standard MTT and colony formation assays, respectively. Changes in expression of 84 critical genes downstream from IGF1R were studied by PCR-array.

Relative to Het1A, IGF1 and IGF2 mRNA were significantly (P<0.05) underexpressed by OE33; by contrast, JHEsoAd1 underexpressed IGF1 but overexpressed IGF2. Administration of either IGF1 (250ng/ml) or IGF2 (500ng/ml) to cell cultures resulted in increased cellular proliferation of JHEsoAD1 cells (after IGF1: 1.24+/-0.05 vs. 1.00+/-0.00 untreated, P<0.01; after IGF2: 1.15+/-0.06 vs. 1.00+/-0.06 untreated, P<0.05). As expected, cell proliferation and clonogenicity of all cell lines were inhibited by CDDP and/or 5-FU (at all dosages ranging from 1-4ug/ml). However, these cytotoxic effects were overcome by the administration of either IGF1 or IGF2 with the exception of: 1) JHEsoAd1 cells treated with CDDP, where IGF2 administration further reduced cellular proliferation (0.35+/-0.06 vs. 0.53+/-0.03 for cells treated with CDDP alone; P<0.05), and 2) JHEsoAd1 cells treated with 5-Fu, where IGF1 administration significantly reduced clonogenicity (0.07+/-0.07 vs. 0.49+/-0.06 for cells treated with 5-Fu alone; P<0.05). PCR-array analysis revealed up-regulation (Het1A and JHEsoAd1) and down-regulation (OE33 and JHEsoAd1) of selected PI3K/AKT pathway genes following IGF1 and IGF2 therapy.

These pre-clinical studies confirm a central role for the IGF axis in esophageal tumor biology, with potential for IGFs to enhance the cytotoxic efficacy of standard chemotherapeutic agents in esophageal cell lines. The identification of novel molecular regulatory pathways downstream from IGF1R modulated by IGF therapy may well inform future cytotoxic and targeting strategies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1092. doi:10.1158/1538-7445.AM2011-1092
Abstract 1094: Role of the CRLF2 ligand TSLP in normal human B-cell development
Authors: Payne Kimberly J. , Milford Terry-Ann , Francis Olivia , Baez Ineavely , Dovat Sinisa , Su Rui-Jun
Published: June 2012

B cell acute lymphoblastic leukemia (B-ALL) is the most common childhood malignancy. Defects that result in overexpression of CRLF2, a cytokine receptor component, have recently been linked with poor outcome in both high-risk and non-high-risk B-ALL. CRLF2, together with the IL-7R alpha chain, forms the signaling receptor for the cytokine TSLP (Thymic Stromal-derived LymphoPoietin). TSLP-induced CRLF2 signaling results in proliferation of murine B cell progenitors, however, the role of TSLP in murine B cell development remains controversial and little is known of its role in human B lymphopoiesis. Human TSLP and CRLF2, in contrast to other cytokine/receptor pairs that act on B cell progenitors, show low homology to their murine counterparts (<40%), lack species cross reactivity, and activate differing intracellular pathways than in mice. Thus, human model systems are essential for understanding the role of CRLF2 function in normal and malignant B lymphopoiesis. We have developed a novel human-only in vitro culture model that allows the study of human B cell development under highly selective cytokine stimulation. This model is based on co-culturing CD34+ hematopoietic stem cells (HSCs) with primary human bone marrow stroma supplemented with selective exogenous cytokines and/or specific cytokine neutralizing antibodies. We have previously used this model to show that IL-7 increases B cell progenitors by over 60 fold and that IL-7 is essential for normal human B cell production (J. Immunol. 2009 82:4255). Here we use our human-only in vitro model to examine the role of TSLP-induced signals in normal human B cell development. Our data show that TSLP induces an expansion of human B cell progenitor populations generated from HSCs in cord blood that is similar in magnitude to that observed for IL-7. A comparison of TSLP and IL-7 effects shows that TSLP targets early B cell progenitors (CD34+) inducing expansion of this population with limited differentiation. In contrast, IL-7 induces rapid differentiation to the CD34- stage followed by proliferation. RT-PCR and ELISA analysis show that normal human bone marrow stoma express both TSLP and IL-7, thus establishing an endogenous source of these cytokines at the site of normal and malignant hematopoiesis. Our data provide evidence that TSLP and IL-7 exert differential and stage specific effects on proliferation and differentiation of B cell progenitors and suggest that deregulation of the complex interplay between these signals in normal differentiation could set the stage for malignant transformation to B-ALL in the presence of cooperative defects in JAK2 and/or Ikaros.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1094. doi:10.1158/1538-7445.AM2011-1094
Abstract 1095: Short Poly A sequence in HGF promoter region is involved in overexpression of HGF in cancer cells
Authors: Sakai Kazuko , Arao Tokuzo , Matsumoto Kazuko , Kimura Hideharu , Fujita Yoshihiko , Kaneda Hiroyasu , Tamura Daisuke , Aomatsu Keiichi , Kudo Kanae , Nagai Tomoyuki , De Velasco Marco A. , Okamoto Isamu , Nakagawa Kazuhiko , Nishio Kazuto
Published: June 2012

Activation of the HGF-MET signaling increases invasion, metastasis and angiogenesis in cancer cells. Therefore, HGF-MET signaling is regarded as an oncogenic signaling pathway, and intensive therapeutic approaches focusing on this signaling pathway are ongoing in the field of cancer treatment. We previously reported that the concentration of serum HGF was strongly related with the treatment outcome of EGFR-tyrosine kinase inhibitors in patients with adenocarcinoma of the lung (Kasahara K. et al, Clin Cancer Res. 2010). Meanwhile, recent study has clearly demonstrated that HGF expression was regulated by short poly A sequence in its promoter region in breast cancer cells (Ma J. et al, J Clin Invest. 2009). In this study, we evaluated the poly A sequence in HGF promoter region and expression levels of HGF in 51 cancer cell lines. The five cell lines exhibited a short poly A sequence (16A-25A) in HGF promoter compared with wild-type (30A). Two of five cell lines overexpressed HGF at mRNA and protein expression levels determined by quantitative RT-PCR and ELISA. On the other hand, there were no HGF-overexpressing cell lines among 46 cell lines with wild-type poly A sequence. Of note, we have also detected the short poly A sequence in cancer cells in patients with lung cancer. Evaluation of short poly A, HGF expression and treatment outcomes of EGFR-tyrosine kinase inhibitors is under way.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1095. doi:10.1158/1538-7445.AM2011-1095
Abstract 1112: Human rhomboid family-1 gene RHBDF1 assists trafficking of TGFa for GPCR-mediated transactivation of EGFR
Authors: Zhou Zhuan , Lu Yi , Deng Weimin , Huang Paulina , Li Lu-Yuan
Published: June 2012

Epidermal growth factor receptor (EGFR) can be transactivated by ligands of G protein-coupled receptors (GPCR). GPCR signaling pathways often involve the formation of clathrin-coated endosomes, which interact with endoplasmic reticulum/Golgi complex to form recycling endosome and secretion vesicles containing EGFR pro-ligands such as pro-TGFα, which is proteolytically processed and released on the cell surface to activate EGFR in an autocrine manner. We showed previously that human rhomboid family-1 gene RHBDF1 is essential for GPCR-EGFR transactivation by assisting the secretion of TGFα, a ligand of EGFR. We show here silencing the RHBDF1 with shRNA leads to inhibition of EGFR ligand-dependent transactivation induced by GPCR ligands such as gastrin release peptide (GRP) and sphingosine-1-phosphate. However, RHBDF1 is not involved in EGFR ligand-independent GPCR-EGFR transactivation induced by isoproterenol. Inhibitors of endosome formation and membrane trafficking such as chlorpromazine, nocodazole, and cytochalasin-D also inhibit GRP-induced EGFR phosphorylation. Inhibitors of endosome acidification such as chloroquine and BFA lessen the inhibitory effect of RHBDF1 shRNA on GRP-induced EGFR phosphorylation. The RHBDF1 protein localizes mainly to the endoplasmic reticulum with pro-TGFα and GRASP55 in starved, quiescent cancer cells, but is phosphorylated and translocated to the cell surface upon GPCR ligands stimulation. GPCR ligands also activate the intracellular signaling molecule Src which then co-translocates with RHBDF1 and clathrin to the plasma membrane. Src-clathrin interaction and co-localization to the cell surface are RHBDF1-dependent. These findings are consistent with the view that, in response to GPCR activation, RHBDF1 assists pro-TGFα trafficking to the cell surface through clathrin coated trafficking.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1112. doi:10.1158/1538-7445.AM2011-1112
Abstract 1114: IGF-II regulates the expression of estrogen receptor beta 5: Role in the survival disparity between African-American and Caucasian women with breast cancer
Authors: O'Bannon Chane’ , Hamilton Nalo , De Leon Daisy
Published: June 2012

African American (AA) women with breast cancer are more likely to be diagnosed at a more advanced stage of the disease, have a higher recurrence rate and a poorer prognosis than Caucasian (CA) women. African American women are more often diagnosed with high-grade breast tumors at an earlier age most of which are negative for the estrogen and progesterone receptors. These factors result in a higher mortality rate among AA patients. This survival disparity can be attributed to socieconomic status and other related factors, however there are biological factors that may also be involved. The insulin-like growth factor II (IGF-II) plays a crucial role in fetal and cancer development by signaling through the IGF-I receptor (IGF-IR), the insulin receptor (InsR) and crosstalk with the estrogen receptor alpha (ER-α). Compared to CA women, AA women have an increased level of proIGF-II expression in breast tissues and higher signaling activation through the IGF-1 and InsRs. Since the estrogen receptor beta (ER-β) is also expressed in a majority of breast cancers we propose that IGF-II may also activate the ER-β signaling pathway. ER-β has 5 isoforms of which ER-β5 is the most abundantly expressed among AA women with breast cancer and it is associated with a poor prognosis and decreased survival. Thus, our hypothesis is that IGF-II activation of the ER-β5 contributes to the survival disparity between AA women and CA women. To test this hypothesis we analyzed the expression of ER-β5 in AA and CA breast cancer cell lines by western blot, siRNA analysis and IGF-II stimulation. IGF-II not only modulated the phosphorylation of ER-β5, but it also promoted the translocation of ER-β5 to different organelles. Thus, we conclude that ER-β5 may contribute to the survival disparity among AA breast cancer patients since ER-β5 impairs the inhibitory effect of other ER-β isoforms.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1114. doi:10.1158/1538-7445.AM2011-1114
Abstract 1121: A novel KIF5B-ALK fusion gene variant with transforming potentials in non-small cell lung cancer
Authors: Wong Daisy Wing-Sze , Leung Elaine Lai-Han , Wong Sunny Kit-Man , Tin Vicky Pui-Chi , Wong Maria Pik
Published: June 2012

BACKGROUND: The anaplastic lymphoma kinase (ALK) gene is frequently involved in chromosomal translocations resulting in fusion genes with different partners found in various lympho-proliferative conditions. In non-small cell lung cancers (NSCLC), the fusion protein encoded by echinoderm microtubule-associated protein-like 4 (EML4)-ALK fusion gene was recently reported to confer oncogenic properties. This study aimed to identify other possible ALK fusion genes and analyze their oncogenic potentials. METHODS: Immunohistochemical analysis was used to screen for aberrant ALK expression in primary NSCLC. 5’ rapid amplification of cDNA ends was used to screen for potential novel 5’ fusion partners of ALK other than EML4-ALK. Reverse transcription polymerase chain reaction and fluorescence in situ hybridization were used to confirm the identity of 5’fusion partner. The genomic breakpoint was verified using genomic sequencing. Overexpression of the novel ALK fusion gene and variants 3a and 3b of EML4-ALK was performed to assess downstream signaling and functional effects. RESULTS: We identified a novel gene resulting from fusion of kinesin family member 5B (KIF5B) exon 15 to ALK exon 20 in a primary lung adenocarcinoma. Western blot of the clinical tumor tissues showed expression of a protein whose size correlated with that of the predicted KIF5B-ALK. Overexpression of KIF5B-ALK in mammalian cells led to STAT3 and AKT activation, enhanced cell proliferation, migration and invasion. These oncogenic properties were comparable to those of EML4-ALK variants. CONCLUSIONS: The discovery of the novel KIF5B-ALK variant with transforming functions further consolidates the role of aberrant ALK signaling in lung carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1121. doi:10.1158/1538-7445.AM2011-1121
Abstract 1127: Lack of MAPK activation in significant number of uveal melanomas with somatic mutation in GNAQ and GNA11
Authors: Abdel-Rahman Mohamed H. , Boru Getachew , Cebulla Colleen M. , Christopher Benjamin , Massengill James B. , Frederick Davidorf H.
Published: June 2012

Purpose: The aims of this study were to investigate the correlation between MAPK activation and somatic mutation in GNAQ and GNA11 genes in uveal melanoma (UM) and to investigate the potential utility of MEK inhibition as a target for therapy of UM in tumors with GNAQ and GNA11 mutations.

Methods: Sequencing and restriction fragment length polymorphism (RFLP) were utilized for detection of activating mutations in codon 209 of the GNAQ and GNA11 genes in 44 primary UMs. The expression of phospho p44/42 MAPK (pERK1/2) was assessed by immunohistochemistry in 44 UMs. In addition, the expression of pERK1/2 and pMEK 1/2 were studied using Western blot analysis in 17 primary UMs (14 with GNAQ or GNA11 mutations) and five UM cell lines (C918, 92.1, OCM3, MEL202, MEL270). Three of the UM cell lines (92.1, MEL202 and MEL270) have GNAQ codon 209 mutations. The effect of three different selective MEK inhibitors (U126, PD98059 and PD184352) on UM cell proliferation and apoptosis were studied.

Results: The majority (30/44, 68.2%) of the primary UM tumors showed somatic mutation in codon 209 of either the GNAQ or the GNA11 genes. With the exception of one tumor, the mutations in either gene were mutually exclusive. Of the 30 UM primary tumors with GNAQ and/or GNA11 mutation, pERK1/2 expression was absent in 26.7%, weak in 33.3%, moderate in 23.3%, and high in only 16.7%. Western blot showed no pMEK1/2 expression in 6/14 (42.9%) of UM with either GNAQ or GNA11 mutations. Two of the UM cell lines with GNAQ mutation showed weak pERK1/2 expression. In addition one showed no pMEK1/2 expression while the other one showed very weak pMEK1/2 expression. Selective MEK inhibitors slowed UM cell proliferation but didn't induce significant apoptosis in UM cell lines regardless of GNAQ status.

Conclusions: Our results indicate that a significant number of UM with somatic mutations in GNAQ or GNA11 showed no MAPK pathway activation. In UM, MEK inhibition is mostly cytostatic suggesting that selective MEK inhibitors alone may not be sufficient to control UM.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1127. doi:10.1158/1538-7445.AM2011-1127
Abstract 1142: A specific miRNA signature characterizes metastasis in renal cell carcinoma
Authors: Heinzelmann Joana , Unrein Andre , Sanjmyatav Jimsgene , Wunderlich Heiko , Grimm Marc-Oliver , Junker Kerstin
Published: June 2012

Background: MicroRNAs (miRNAs) as small non-coding RNAs are regulators of gene expression in tumor development and progression. However, its specific role in metastasis of clear cell renal cell carcinoma (ccRCC) is almost unknown. Therefore, we aimed in defining specific miRNA expression pattern of metastatic ccRCC and identifying cellular functions of specific miRNAs deregulated in metastatic process.

Methods: We isolated total RNA of 29 ccRCC samples (including 13 non-metastatic and 16 metastatic tumors) and from 16 metastases using snap frozen tissue. Single channel microarray analysis was performed for a global miRNA expression profiling in 22 ccRCC samples. The validation of the differentially expressed miRNAs in metastatic ccRCC compared to non-metastatic tumor samples and furthermore in metastases of ccRCC was performed by using qRT-PCR.

Results: We identified a miRNA expression pattern that distinguishes between metastatic and non-metastatic ccRCC. Furthermore, we found a group of miRNAs with altered expression in early-metastatic compared to late metastatic and non-metastatic tumors. These microarray results were verified by qRT-PCR. By analyzing the expression of four of these differentially expressed miRNAs (26a, 30c, 126, 451) in metastases tissue by qRT-PCR we detected the same expression behavior as in the primary metastatic tumor counterparts.

Conclusion: Our findings indicate that miRNAs are involved in the metastatic process. Thus, we were able to identify a specific miRNA expression pattern in metastatic ccRCC. Metastases of ccRCC are characterized by the same expression value of specific miRNAs as the primary metastatic tumor tissue. These investigations showed that specific miRNAs are possible prognostic markers for metastatic disease in RCC patients. Further studies have to define the functional role of the miRNAs in the metastatic process.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1142. doi:10.1158/1538-7445.AM2011-1142
Abstract 1147: microRNA expression profiling in cisplatin resistant human non-small cell lung cancer cell lines
Authors: Suzuki Toshihiro , Irisawa Ai , Nishio Kazuto , Togawa Tadayasu , Sakuraba Hitoshi
Published: June 2012

Cisplatin combination chemotherapy is one of the important treatments for many cancers including lung cancer. Despite impressive initial clinical responses, the majority of patients eventually develop some degree of resistance to cisplatin-based therapy. To understand the clinically relevant mechanisms of resistance, many studies have been aimed at clarifying the biochemical/molecular alterations of cisplatin-resistant cells. However, cellular resistance mechanisms of cisplatin are still unclear. MicroRNA (miRNA) is small noncoding RNA that function to control gene expression. These small RNAs have been shown to contribute to the control of cell growth, differentiation and apoptosis, important features related to cancer development progression and sensitivity of chemotherapy. However, the interaction between the development of cisplatin resistance and miRNA has not been previously explored.

In this study we utilized a miRNA array to compare the differentially expressed miRNAs in cisplatin-resistant cell lines, PC-9/CPr and PC-14/CPr and their parental cell lines, PC-9 and PC-14. We verified that miR-7, miR-106 and miR-181 were up-regulated in cisplatin-resistant non-small cell lung cancer cells by means of real-time PCR and discusses the potential of these miRNAs as biomarkers for cisplatin sensitivity in combination chemotherapy. We further investigated the role and mechanisms of miR-181 in cisplatin resistance. We found that specific miR-181 precursor (Pre-miR-181) transfection significantly acquired cisplatin resistance.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1147. doi:10.1158/1538-7445.AM2011-1147
Abstract 1148: Development and validation of a second generation microRNA-based assay for diagnosing tumor tissue origin
Authors: Rosenwald Shai , Edmonston Tina , Barshack Iris , Feinmesser Meora , Huszar Monica , Mueller Wolf , Fogt Franz , Spector Yael , Aharonov Ranit , Meiri Eti
Published: June 2012

Background: Cancer of unknown primary (CUP) constitutes 3%-5% of all newly diagnosed cancer cases. It presents a major diagnostic challenge due to the therapeutic implications of the tissue origin of the cancer. MicroRNAs are a family of non-coding, regulatory RNA genes that are involved in development, differentiation and cell cycle and were shown to be involved in oncogenesis. MicroRNAs are highly tissue-specific and therefore ideal candidates for CUP diagnosis. We have previously presented a biologically-motivated classification assay to identify the tissue of origin using a set of 48 microRNAs measured on a qRT-PCR platform. This assay can differentiate 25 different tumor types from 17 origins. We present here the development and validation of a second generation assay that can identify 42 different tumor types using a custom array platform.

Methods: Over 1200 primary and metastatic tumor FFPE samples were used for the training and development of the assay. High-quality RNA, including the well-preserved microRNA fraction, was extracted from the FFPE blocks using proprietary protocols. Expression levels of all known and additional Rosetta's proprietary microRNAs were profiled using a custom array platform. A combination of a decision tree classifier with a KNN classifier was developed to identify the tissue of origin based on the microRNA expression patterns. A validation set of more than 500 independent FFPE samples of primary tumors and metastases was analyzed blindly and classified using the developed assay.

Results: The assay is able to identify 42 tumor types based on the expression of only several dozens microRNAs with very high accuracy. Based on the validation set, the sensitivity (positive agreement) for identifying the tumor origin is 85%. In more than 80% of the cases, a single answer was reported, and for these cases, the sensitivity is 90%.

Conclusions: Previous studies have highlighted the tissue-specificity of microRNA expression, and have demonstrated their potential use for classification of human malignancies. An assay utilizing microRNA expression in paraffin-embedded tissue for identification of the tissue of origin was previously developed. Here we present the development and the performance of a second generation assay, designed to identify 42 tumor types. This assay will provide an important new tool for diagnosing tumor tissue origin.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1148. doi:10.1158/1538-7445.AM2011-1148
Abstract 1153: Nanoparticle delivery of microRNA-107 reduces the tumorigenicity of head and neck squamous carcinoma (HNSCC)
Authors: Piao Longzhu , Zhang Manchao , Su Tizhi , Bhave Sanjay , Li Hong , Teknos Theodoros N. , Pan Quintin
Published: June 2012

Head and neck squamous cell carcinoma (HNSCC) is the sixth most prevalent cancer worldwide with about 600,000 new cases diagnosed in the last year. The 5-year survival rate of HNSCC has remained stagnant despite advances in the clinical management of this disease. Understanding the molecular pathways that lead to aggressive HNSCC is crucial to identify new “druggable” targets for anti-cancer drug development. Recently, our laboratory showed that miR-107 is significantly reduced in primary HNSCC tumors and HNSCC cell lines. Ectopic expression of miR-107 in CAL-27 HNSCC cells significantly inhibited cell proliferation, DNA replication, clonogenic survival, and tumor growth in nude mice. Our data demonstrate the potential application of miR-107 as a novel anti-cancer therapeutic for HNSCC. In this study, we determine the efficacy of miR-107 encapsulated nanoparticles (NP-miR-107) in a xenograft model of HNSCC. NP-miR-107 was successfully formulated by ethanol injection method. The particle size of NP-miR-107 was 47.8 ± 3.5 nm and zeta potential was +12.1 mV. NP-miR-107 significantly retarded (57.6% inhibition, p<0.02) tumor growth of CAL27 HNSCC tumors compared to NP-miR-control; tumor volume of 340.6 ± 51.6 mm3 and 804.1 ± 181.6 mm3 for the NP-miR-107 group and the NP-miR-control group, respectively. Moreover, Kaplan-Meir analysis showed a survival advantage for the NP-miR-107 treatment group (p=0.039). Our results demonstrate that our nanoparticles are an effective carrier approach to deliver therapeutic miRs to tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1153. doi:10.1158/1538-7445.AM2011-1153
Abstract 1165: PKC epsilon modulated miRNAs profiling during development of prostate cancer in transgenic adenocarcinoma of mouse prostate model
Authors: Hafeez Bilal B. , Zhong Weixiong , Verma Ajit K.
Published: June 2012

MicroRNAs (miRNAs) are small nucleotides endogenously expressed non-coding RNAs that have been implicated in a wide range of cellular processes. Dysregulation of miRNA expression has been identified in a number of cancers including prostate. Evidence indicates that miRNAs are involved in prostate cancer (PCa) development and metastasis via activating several oncogenes and inhibiting tumor suppressor genes. However, the molecules’ targeting these microRNAs is poorly understood. Protein kinase C epsilon (PKC∈), a novel PKC isoforms, is over-expressed in both human and mouse PCa. We have previously shown that PKC∈ expression correlates with the aggressiveness of PCa. We generated PKC∈-deleted TRAMP mice by crossbreeding transgenic adenocarcinoma of mouse prostate (TRAMP) (TG//WT) mice with PKC∈ knock out (TG//KO) mice. Sixteen weeks old TG Wild-type (TG//WT), TG//PKC∈ Heterozygous (TG//Het) and TG//PKC∈ KO (TG//KO) mice were evaluated for PCa development and metastasis. We observed that deletion of PKC∈ in TRAMP mice inhibited PCa development and metastasis as observed by CT-PET scanning, tumor weight and histopathology of livers, lungs, kidneys and lymph nodes. To determine the clues about the miRNAs, which are modulated by PKC∈, we performed global miRNA profiling in prostate tissues of TG//WT and TG//KO mice by using miRNA Microarray chip containing transcripts of 750 miRNAs (Listed in Sanger miRBase Release 15.0). We observed a significant down-regulation of 7 miRNAs (miRNA762, miR125b-5p, miR-16, miR-467a-1, miR-574-3p, miRNA-451 and miR-669f) and up-regulation of 6 miRNAs (miR-1187, miR-1195, miR-2133, miR-574-5p, miR-690 and miR-709) in prostate tissues of TG//KO mice compared to TG//WT mice. It has been reported that miRNA-451 plays a role in the induction of mTOR signaling, which is well documented for its role in human PCa progression. Importantly, we observed 20-fold decreased expression of miRNA-451 in prostate tissues of TG//KO mice compared to TG//WT mice. It has been reported that up-regulation of miRNA-709 inhibits expression of BORIS, a tumor promoting transcription factor. We observed two fold induction of miRNA-709 in TG//KO mice. Taken together these finding suggest that PKC∈ plays an important role in PCa development and metastasis via up-regulating oncogenic miRNA and down regulating tumor suppressing miRNA. We suggest that these miRNAs modulated by PKC∈ have role in PCa development and metastasis. However further studies warranted to understand the functional implication of these miRNAs in human PCa. (Support: NIH Grant, CA35368).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1165. doi:10.1158/1538-7445.AM2011-1165
Abstract 1179: Global increase in ultraconserved non-coding RNA expression in pancreatic adenocarcinoma
Authors: Azevedo Ana Clara P. , Jiang Jinmai , Lee Eun Joo , Gusev Yuriy , Allard David , Tuveson David A. , Calin George A. , Schmittgen Thomas D.
Published: June 2012

Transcribed ultraconserved regions (T-UCRs) are a class of 482 non-coding RNAs with 100% sequence conservation among human, rat and mouse genomes. T-UCRs are differentially expressed in colorectal cancer, leukemia and neuroblastoma, however their expression in pancreatic adenocarcinoma (PDAC) has not been studied. We used qPCR to profile all 482 T-UCRs in a variety of pancreatic cancer specimens including pancreatic adenocarcinoma, chronic pancreatitis and benign/normal pancreas and in the pancreas of genetically engineered mice (P48+/Cre;LSL-KRASG12D and PDX-1-Cre;LSL-KRASG12D). Of the 257 expressed T-UCRs in the human pancreas tissues, 166 were differentially expressed in the PDAC compared to normal and adjacent benign pancreas (≥ 1.5 fold; p < 0.05). In the genetically engineered mice, 29% of the expressed T-UCRs were differentially expressed in the pancreas of old P48+/Cre;LSL-KRASG12D mice (mean age, 231 days) compared to control. In both the patient and mouse data, the vast majority of the differentially expressed T-UCRs increased with disease progression. Interestingly, the expression pattern among groups of two T-UCRs closely correlated with each other. This was to be expected for T-UCRs directly adjacent to one another in the genome, but surprisingly we found strong correlations among T-UCRs that are located on different chromosomes. This suggests that large groups of T-UCRs are co-expressed in PDAC. In vitro models that simulate the desmoplastic reaction have been shown it to alter gene expression in PDAC. In an effort to understand the mechanism responsible for this global increase in expression in PDAC, the T-UCRs were profiled in normal pancreas (HPDE) cells that were cultured on the extracellular matrix deposited by Panc-1 cells. Ninety-one T-UCRs were increased in the HPDE co-cultures (fold-change > 1.5, p< 0.05). We report a global increase in the T-UCR expression in PDAC tissues, the pancreas of genetically engineered mice and in HPDE cells co-cultured with Panc-1 cells. The ability of the MAP kinase and TGF-β signaling pathways to regulate T-UCR expression will be investigated in the future.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1179. doi:10.1158/1538-7445.AM2011-1179
Abstract 1180: Activation of antisense transcription by Top1cc in human colon cancer cells
Authors: Marinello Jessica , Bertozzi Davide , Bueno Susana , Iurlaro Raffaella , Sordet Olivier , Chillemi Giovanni , Capranico Giovanni
Published: June 2012

Camptothecin (CPT) and derivatives are effective anti-cancer agents exerting their activity through the inhibition of DNA topoisomerase I (Top1). The cascade of events following the collision of a replication fork with CPT-trapped Top1 (TopIcc) has been deeply studied. Recently, strong evidence demonstrated that Top1cc affects specific transcriptional mechanisms as well. In particular, Top1ccs specifically favor RNA polymerase II escape from promoter-proximal pausing sites, and can alter chromatin structure and accessibility. We have also demonstrated that the CPT-induced transcriptional stress determines a changed balance between sense and antisense transcripts at the human HIF-1α locus in cancer cells [Capranico et al., BBA 2010]. Here, we have further investigated the Top1cc-related antisense transcriptional activation at the single gene locus and genome-wide. CPT treatments promote the transcription, in normal as well as hypoxic conditions, of a novel long non-coding antisense RNA at the 5’ of the human HIF-1α mRNA (5’aHIF1α), which is 5’-capped and polyadenylated, indicating that is transcribed by RNA polymerase II. The 5’aHIF1α RNA localizes preferentially in the nuclear compartment, in particular RNA FISH experiments show that it accumulates mainly in the perinuclear region of human HCT116 cells. Thus, the data may suggest that it may have a regulatory role in membrane RNA/protein trafficking. To investigate whether the described effect at HIF-1α locus is a general event after Top1cc, we have carried out a genome-wide analyses of cellular transcripts. rRNA-depleted total RNA samples from CPT-treated and untreated HCT116 cells were subjected to bisulfite C>T conversion to maintain the information of the strand polarity of transcripts, and then sequenced on the Illumina platform (Solexa). Initial analyses of the ratio between sense and antisense sequence tags indicate that Top1cc induces the activation of antisense transcription at several gene loci. The results thus indicate that activation of antisense transcription may be a general response to the transcriptional stress caused by Top1ccs, possibly contributing to the pharmacological activity of camptothecins.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1180. doi:10.1158/1538-7445.AM2011-1180
Abstract 1193: Guanylate binding protein-1, a novel oncogenic gene in glioblastoma
Authors: Li Ming , Cavenee Webster , Furnari Frank
Published: June 2012

Glioblastoma multiforme (GBM) is the most common and deadly primary malignant brain tumor in adults. The most frequent genetic lesion associated with GBM, found at 40-60% frequency, is amplification and mutation of the EGFR gene. Here we report that guanylate binding protein-1 (GBP1), an immune response large GTPase, is involved in EGFR-mediated glioblatoma invasion through the modulation of matrix metallopeptidase-1 (MMP1) expression and show that GBP1 is upregulated in human GBM tumors relative to normal brain tissues, especially in EGFRvIII-positive samples. We further show that EGFRvIII promotes GBP1 expression at the transcriptional level through stimulation of the p38 MAPK signaling pathway and targeting GBP1 by lentivirus shRNA transduction significantly reduces EGFRvIII-mediated tumorigenicity of orthotopic xenografts in the mouse. Reciprocally, overexpression of GBP1 alone in glioma cells was sufficient to enhance glioma tumorigenesis by increasing cell proliferation, angiogenesis and decreasing cell apoptosis. Taken together, these results reveal that GBP1 is a novel oncogenic gene downstream of EGFRvIII signaling and suggest that GBP1 may serve as a promising therapeutic target for the treatment of GBM patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1193. doi:10.1158/1538-7445.AM2011-1193
Abstract 1224: Repression of cancer cell senescence by PKCι
Authors: Restall Ian J. , Paget Judith A. , Daneshmand Manijeh , Amin Md. Shahrier , Islam Shahidul , Lorimer Ian A.J.
Published: June 2012

Senescence is an irreversible growth arrest phenotype adopted by cells that has a key role in protecting organisms from cancer. There is now considerable interest in therapeutic strategies that reactivate this process to control the growth of cancer cells. Protein kinase C iota (PKCι) is a member of the atypical protein kinase C family and an important downstream mediator in the phosphoinositide pathway. PKCι expression was found to be upregulated in a subset of breast cancers and breast cancer cell lines. Introduction of mutant, oncogenic PIK3CA, but not wild-type PIK3CA, into breast mammary epithelial cells increased both the expression and activation of PKCι. In breast cancer cell lines overexpressing PKCι, depletion of PKCι increased the number of senescent cells, as assessed by senescence-associated β-galactosidase, morphology and bromodeoxyuridine incorporation. This phenomenon was not restricted to breast cancer cells, as it was also seen in glioblastoma cells. Senescence induction did not require p53, p16 or Arf and was not associated with activation of the DNA damage response. Depletion of PKCι had no effect on senescence in normal mammary epithelial cell lines. We conclude that PKCι is overexpressed in a subset of cancers where it functions to suppress premature senescence. This function appears to be restricted to cancer cells and inhibition of PKCι may therefore be an effective way to selectively activate premature senescence in cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1224. doi:10.1158/1538-7445.AM2011-1224
Abstract 123: miR-145 function as a tumor suppressor targeting multiple oncogenes in prostate cancer
Authors: Fuse Miki , Nohata Nijiro , Mutallip Muradil , Kikkawa Naoko , Sakamoto Shinichi , Kojima Satoko , Chiyomaru Takeshi , Kawakami Kazumori , Enokida Hideki , Nakagawa Masayuki , Ichikawa Tomohiko , Seki Naohiko
Published: June 2012

Prostate cancer remains one of the most prevalent cancers and a major cause of morbidity and mortality in the western world. Disease progression and the development of hormone refractory disease remain major causes of cancer-related death. The understanding of the new molecular pathways of hormone refractory would be helpful in improving diagnosis and therapy of the disease.

microRNAs (miRNAs) are small, non-coding RNAs with a length of approximately 22 nucleotides. They regulate gene expression by promoting mRNA cleavage and at the posttranscriptional level by translational suppression. They play important roles in various biological and metabolic processes, including development, differentiation, signal transduction, cell maintenance, and cancer. Bioinformatic predictions indicate that miRNAs regulate more than 30% of the protein coding genes. It is estimated that approximately 1,000 miRNAs exist in the vertebrate genome.

An important role for miRNAs in the development of cancer has emerged in recent years. miRNAs are aberrantly expressed in many human cancers, and they may function as oncogenes or tumor suppressors. Up-regulated miRNAs could function as oncogenes by negatively regulating tumor suppressor genes, while down-regulated miRNAs could act as tumor suppressors, inhibiting cancers by regulating oncogenes.

Down-regulation of miR-145 has been reported in many types of human cancer, including prostate cancer, suggesting that miR-145 function as a tumor suppressor. In this study, we focused on target identification of miR-145 in prostate cancer, based on gene-wide gene expression analysis of miR-145 transfection cells. According to our expression analysis showed that down-regulated genes (FSCN1, UNG and EIF4EBP2) were identified. Loss-of-function assays revealed that FSCN1 inhibited cell proliferation, migration, and invasion in PC3 and DU145 cells, suggesting FSCN1 function as an oncogene.

The identification of tumor suppressive miRNA and their target genes could provide new insights into potential mechanisms of prostate carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 123. doi:10.1158/1538-7445.AM2011-123
Abstract 1235: Induction of a p21 mediated senescence program by p53 impairs the apoptotic response to chemotherapy and clinical outcome in breast cancer
Authors: Jackson James G. , Chang Leslie L. , Quintás-Cardama Alfonso , El-Naggar Adel K. , Lozano Guillermina
Published: June 2012

How p53 mediates response to DNA damage induced by chemotherapy treatment of breast cancer is not well understood with few mechanistic studies addressing the conflicting reports in the literature. We have used the MMTV-Wnt1 mouse model of breast cancer to examine the role of the p53 pathway in the biochemical, cellular and clinical response to doxorubicin. We found that treatment of p53 wild type breast tumors, or heterozygous tumors that retained the wild type allele, resulted in an initial mean tumor regression of only 35%, followed by stable disease (∼1 to 2 weeks) and relapse by 11 days. Surprisingly, we found that breast tumors with mutant p53 responded better to treatment, with regression of 70% and 24 days to relapse (p<0.001). Investigating the mechanism of these responses, we found that p53 mutant tumors harvested 24 hours after final treatment did not induce p53 targets after treatment when compared to untreated tumors, did not enter proliferative arrest, but did undergo p53-independent apoptosis, consistent with tumor regression data. Conversely, treated p53 wild type tumors induced p53 targets such as p21 and Puma, were negative for the proliferation marker Ki67, and did not induce apoptosis. Further characterization of the partial response in p53 wild type tumors revealed that tumors harvested in the stable arrest phase (7-14 days after treatment) had elevated markers of senescence, including p21, Dec1, Dcr2, p15, p16 and PML, compared to relapsed or resistant tumors. To more precisely address which of the myriad p53 functions was responsible for the poor response in p53 wild type tumors, we bred the MMTV-Wnt1 mice into a p21 null background and treated and monitored mice that formed tumors. We found that genetic loss of p21 in the tumors was sufficient to achieve the dramatic response observed in p53 null tumors. These data show that p53 mediated activation of a senescence program via p21 is the primary outcome in treated MMTV-Wnt1tumors, resulting in an incomplete response, stable disease and eventual early relapse, while treated p53 mutant or p21 null tumors undergo cell death and tumor regression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1235. doi:10.1158/1538-7445.AM2011-1235