bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4082: U6 upregulation in cancer
Authors: Hwang Larn , Lin Xiaoqin , Desai Neil , Trieu Vuong 
Published: February 2011

Abstract
Background: U6 snRNA is a non-coding RNA that is a component of the splicesome, which is involved in RNA splicing of the pre-mRNA. In view of the critical function performed by U6 snRNA in RNA splicing and therefore cellular proliferation, it is possible that U6 would play a role in cancer development. In this study, U6 levels were examined across various tumor types.
Methods: Detection of U6 snRNA in various normal and tumor tissue was performed with Tissue Microarray by In Situ Hybridization technique. The 5′-DIG-labeled U6 LNA probe (sequence 5′-CACGAATTTGCGTGTCATCCTT-3′) from Exiqon was used for this study and the Vector Red was used as substrate. The U6 stained tissue microarray was then scanned by GenePix 6.0 (Molecular Devices) at 532 nm to detect the staining intensity from each tissue sample. Each sample on the TMA was also visually scored by the intensity on a scale of 0 to 4. A quantitative real-time PCR assay was also used to confirm the overexpression of U6 snRNA in breast and other cancers.
Results: A baseline study using tumor microarrays and visual scoring suggested that U6 snRNA levels were increased in tumors relative to normal tissues. This was confirmed using the rapid U6 snRNA ISH for colon carcinoma, melanoma, pancreatic carcinoma, renal carcinoma, and lung carcinoma. In all cases, U6 levels were significantly elevated in tumor versus normal by t-test. By RT-PCR, U6 snRNA levels were also found to be increased relative to the corresponding normal tissue in carcinoma of the breast, colon, kidney, liver, lung, and ovary. No increase over the corresponding normal tissue was found in prostate and thyroid carcinomas. In breast carcinoma, there was no correlation between U6 snRNA levels and estrogen receptor status, progesterone receptor status or erb-B2 expression levels.
Conclusions: Using a scanning system and tumor tissue array, we were able to demonstrate the use of microarray scanner to quantitate ISH signal. Application of this technology to U6 snRNA demonstrated that U6 is significantly elevated in most cancer types examined.
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 4082.
Abstract 4083: Long stress-responsive non-coding transcripts identified in a whole genome screen have altered expression in cancer
Authors: Silva Jessica M. , Perez Damon S. , Pritchett Jay , Halling Merideth , Smith David I. 
Published: February 2011

Abstract
The number of non-coding transcripts (NCTs) exceeds the number of transcripts that code for protein. Their diverse and important functions are now being identified and utilized in the lab as well as in the clinic. Utilizing the Affymetrix Gene Chip Human Tiling 1.0R Array Set, we analyzed the transcription across the entire genome in both normal human bronchial epithelial cells (NHBE) alone and cells exposed to the tobacco carcinogen (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone), NNK. We focused our efforts on the identification and function of novel long non-coding transcripts that had increased expression after exposure to NNK. We report on the identification of 12 long NNK-induced non-coding transcripts (NiTs) that are 300 nucleotides and larger in length. We first validated that these transcripts were indeed stress responsive using quantitative real-time RT-PCR (qPCR) and by Northern blot analysis revealed that these transcripts were between two and four kilobases in length. We also show using qPCR that these transcripts are more abundantly expressed in normal rapidly growing tissues or tissues that are under cellular stress. We show that there is altered expression of NiTs in a panel of lung cancer cell lines and many of these transcripts had increased expression in many breast cancer cell lines. We further analyzed one of these transcripts, NiT5 and identified it to be a 2.5 kb antisense, polyadenylated transcript by RLM-RACE and by Northern Blot analysis. Isolating nuclear and cytoplasmic fractions in BT474 cells, we identified Nit5 to be localized in the nucleus and found it to be transcribed by RNA polymerase III. NiT5 also has increased expression in breast, ovarian and cervical cancer-derived cell lines relative to normals from these three sites, but not increased expression in lung cancer cell lines. In summary, our results suggest that these novel long non-coding transcripts play an important role in stress also may play a role in cancer.
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 4083.
Abstract 410: Expression of class 3 semaphorins in prostate cancer and modulation by hypoxia
Authors: Williamson Magali , Blanc Veronique , Nariculam Joseph , Freeman Alex , Masters John 
Published: February 2011

Abstract
Class 3 semaphorins are secreted proteins that act as guidance cues for migrating cells via their transmembrane receptors, plexins. Semaphorins have a role in cancer affecting tumour growth both directly and indirectly, by affecting angiogenesis. The aims of this study were to determine if semaphorins have a potential role in prostate cancer and if their expression is regulated by hypoxia.
Here we show that semaphorins and their transmembrane receptors, plexins and neuropilins, are widely expressed in prostate cancer cell lines and tissue with a significant overexpression of Sema3E in tumour tissue.
Co-expression of semaphorins and their receptors by tumour cells suggests that semaphorins may act through an autocrine or paracrine mechanism in prostate cancer cells and have a direct effect on prostate tumour progression. We found that Sema3E affected integrin-mediated adhesion to fibronectin of prostate cancer cells, either enhancing or inhibiting adhesion, depending on cellular context. Sema3E also inhibited the migration of prostate cancer cells.
As well as affecting tumour progression directly, class 3 semaphorins act as angiogenesis factors. We found that expression of several semaphorin genes is affected by growth in hypoxic conditions. Notably, expression of sema3C is upregulated and Sema3A and Sema3E are down regulated by hypoxia in prostate and colon cancer cells. The down-regulation of sema3A and 3E by hypoxia is consistent with a role for Sema3A and 3E as anti-angiogenic factors in prostate cancer. These results indicate a potential role for Sema3E in prostate cancer.
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 410.
Abstract 4101: Fibroblast growth factor receptor (FGFR) 2 IIIc, is a novel molecular target for suppressing pancreatic cancer cell growth and migration
Authors: Ishiwata Toshiyuki , Matsuda Yoko , Kawahara Kiyoko , Yamamoto Tetsushi , Teduka Kiyoshi , Uchida Eiji , Korc Murray , Naito Zenya 
Published: February 2011

Abstract
Background: Fibroblast growth factor receptor (FGFR) isoforms IIIb and IIIc arise as a consequence of alternative splicing of the carboxy-half of the third immunoglobulin (Ig)-like domain, with IIIb isoforms being mainly expressed in epithelial cells and IIIc isoforms expressed mainly in mesenchymal cells. In pancreatic ductal adenocarcinoma (PDAC), the FGFR1 IIIb isoform mediates inhibition of cancer cell proliferation, migration, and invasion, while FGFR1 IIIc mediates cancer cell proliferation. We have previously reported that the FGFR2 IIIb isoform, also known as keratinocyte growth factor receptor/KGFR, is expressed in PDAC, and its expression correlates with VEGF-A expression and venous invasion. Furthermore, there was a significant correlation between co-expression of FGFR2 IIIb and KGF, and a poor therapeutic response. Single nucleotide polymorphisms (SNPs) of the FGFR2 gene are associated with breast and endometrial cancers, and missense mutations or copy number gains of the FGFR2 gene occur in breast and gastric cancers. A class switch of FGFR2IIIb to FGFR2IIIc is reportedly associated with a more-malignant phenotype in prostate and bladder cancers. Accordingly, in the present study we examined the role of FGFR2 IIIc in PDAC. Methods: FGFR2 IIIc expression levels were examined in six pancreatic cancer cell lines using real-time PCR and western blot analyses. Immunohistochemical analysis using anti-FGFR2 IIIc antibodies was performed on 51 pancreatic cancer patients, and FGFR2 IIIc mRNA expression was confirmed by in situ hybridization analysis. FGFR2 IIIc stably transfected KLM-1 cells were prepared and the growth rates in vitro and in vivo were compared with mock cells. The effects of specific anti- FGFR2 IIIc antibodies on the growth and migration of PANC-1 cells were also examined. Results: FGFR2 IIIc was localized in cancer cells in 37 of 51 PDAC samples (73%), and the expression correlated with retroperitoneal invasion. FGFR2 IIIc expression was detected in all six pancreatic cancer cell lines, and the expression level was high in PANC-1 cells and low in KLM-1 cells. The growth rates of FGFR2 IIIc stably transfected KLM-1 cells were higher than those of mock cells in vitro and in the subcutaneous tissue of nude mice, whereas anti-FGFR2 IIIc antibodies inhibited the growth and migration of PANC-1 cells. Conclusion: These findings indicate that FGFR2 IIIc is commonly overexpressed in PDACs, where it promotes cancer cell growth and migration. Thus, FGFR2 IIIc may serve as a novel therapeutic target in PDAC.
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 4101.
Abstract 4104: Protease-activated receptor 2, a KLK14 target in colon cancer cells
Authors: Gratio Valérie , Loriot Céline , Oikonomopoulou Katerina , Chung Hyunjae , Diamandi Eleftherios P. , Hollenberg Morley D. , Darmoul Dalila 
Published: February 2011

Abstract
Serine proteinases signal through proteinase-activated receptors (PARs). Our studies have implicated PAR1 and PAR4 (thrombin receptors) and PAR2 (trypsin receptor) in human colon cancer growth. However, endogenous activators of PARs in colon tumors are still unknown. Recently, members of the kallikrein-related peptidase (KLK) family have been shown to activate PARs in many cell systems. Thus, our aim was to determine if KLK14, a KLK family member, is expressed in colonic tumors and to see if this KLK can activate PARs in human colon cancer cells. The presence of KLK14 in colon cancer cell lines was assessed by RT-PCR, ELISA and immunofluorescence, and its expression in human colonic tissue was assessed by immunohistochemistry. The effects of recombinant KLK14 in parallel with PAR1/2 agonists (SFLLRN-NH2, an activating peptide for both PAR1 and PAR2; TFLLR-NH2, a selective activating peptide for PAR1 and the selective PAR2 agonists, 2-furoyl-LIGRLO-NH2; SLIGRL-NH2) were assessed for calcium flux, receptor internalization and ERK1/2 phosphorylation in a human colon cancer-derived HT29 cell line that expresses PAR1 and PAR2. We found that: a) KLK14 mRNA (RT-PCR) was present in 16 out of 16 human colon cancer cell lines, b) KLK14 protein is present (ELISA) in HT29 conditioned media and other colon cancer cell lines; c) there was strong staining of KLK14 in the cytoplasmic compartment of HT29 cells and of human colonic tumors (immunofluorescence and immunohistochemistry; paraffin sections) d) KLK14 (0.1 µM) caused prompt increases in intracellular calcium ([Ca2+]i) in HT29 cells (Microspectrofluorimetry with Fura 2/fluo-3). The KLK14-induced Ca2+-flux was abrogated after an initial challenge of the cells with SFLLRN-NH2 (100 µM), which desensitizes PAR1 and PAR2 simultaneously. Desensitization with 2-furoyl-LIGRLO-NH2 (10 µM) a potent PAR2 agonist, attenuated most of the Ca2+ flux induced by KLK14, whereas responses to TFLLR-NH2, the PAR1 agonist, were minimally affected. Consistently, a first challenge with thrombin did not affect KLK14-induced Ca2+ flux. These results strongly suggest that KLK14 activates preferentially PAR2 in HT29 cells. e) KLK14 initiates a dramatic loss of cell surface PAR2 (microscopic analysis) due to its internalization and f) KLK14 induces a rapid (within 5-10 min) and significant ERK1/2 phosphorylation in HT29 cells. This KLK14-PAR signaling pathway may represent a novel therapeutic target for colon tumorigenesis.
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 4104.
Abstract 4106: Methods for the development of carcinoid cell lines and carcinoid associated fibroblasts
Authors: Eric Sceusi L. , Rashid Asif , Zhou Yunfei , Samuel Shaija , Fan Fan , Xia Ling , Ye Xiang-Chang , Lu Jia , Tozzi Federico , Gaur Puja , Yao James , Meyn Raymond , Ellis Lee M. 
Published: February 2011

Abstract
Background: Research in carcinoid and neuroendocrine tumors (NETs) is limited by the lack of existing midgut derived cell lines. The two existing NET cell lines, BON and H727, are foregut derived. Foregut NETs are phenotypically distinct from midgut NETs, thus the development of a midgut derived NET cell line is of critical importance. Human NETs contain areas of stroma that likely effect carcinoid cell growth. In other types of cancer, cancer associated fibroblasts (CAFs) have been shown to secrete factors that mediate tumor cell growth. In our preliminary studies, cultured human NET cells showed better attachment to plates when they were in close association with fibroblasts. Unfortunately, the relatively slow growth of NET cells allows fibroblast overgrowth and ultimately detachment of the cancer cells. We have developed a novel approach to processing and culturing NET cells through the isolation of autologous CAFs and their use as irradiated feeder layers.
Methods: Human NET surgical specimens are procured under sterile conditions and processed in our lab. The tissue is digested to a single cell suspension. Anti-fibroblast labeled beads (Miltenyi Biotec) are used to separate CAFs from tumor cells. CAFs are then cultured and irradiated to produce feeder layers for the tumor cells. The NET cells are initially maintained on extracellular matrix protein coated plates and then plated on top of the CAF monolayer.
Results: CAFs in culture can survive 40 Gy of irradiation and will not proliferate after this treatment. These cells were viable at 12 days, at which point they were split. These irradiated fibroblasts were able to reattach to standard culture flasks and were viable, but did not proliferate after splitting, while the non-irradiated CAF controls had reached confluence by day 12 of this second passage. We currently have 5 patient NET samples that have been processed and plated in vitro on autologous CAFs. NET cells can then be separated from the irradiated fibroblasts using standard trypsinization followed by fibroblast separation using anti-fibroblast beads as described above. The tumor cells can then be used for analysis or further culturing. Early observations shows that the cancer cells appear more elongated and flattened when adjacent to fibroblasts, possibly indicating better attachment and survival.
Conclusions: While NET cells remain difficult to culture, we have developed a system that utilizes autologous CAFs in our efforts to potentially establish new NET cell lines for in vitro and in vivo studies.
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 4106.
Abstract 4108: Syndecan-1 overexpression enhances colon cancer growth by modulating heparan sulfate binding growth factors sensitivity
Authors: Hayashi Hiroaki , Minamiguchi Kazuhisa , Kanoh Akira , Ishida Keisuke , Terada Tadashi , Matsuo Kenichi 
Published: February 2011

Abstract
Syndecan-1 is a multifaceted transmembrane heparan sulfate proteoglycan known for its significant role in a wide range of cellular processes by interacting with a multitude of extracellular components. Although loss of syndecan-1 correlates well with the gain of cancerous characteristics in a wide range of cancers, increased expression of syndecan-1 also corresponds to adverse outcomes in some cancers. The biological function is yet to be completely understood in most of malignancies, including colon cancer. In the present study, to evaluate biological functions of syndecan-1, its expression level was altered, and subsequent outcomes were examined using human colon cancer LoVo cells. Ectopic expression of syndecan-1 in LoVo cells (LoVo/SDC1) led to larger tumor formation than did control LoVo cells (LoVo/mock) in mice. Lower concentrations of HB-EGF and HGF could activate their receptors in LoVo/SDC1 than in LoVo/mock. And, the prolonged phosphorylation of EGFR induced by HB-EGF was observed in LoVo/SDC1. Furthermore, HB-EGF more effectively promoted cell growth of LoVo/SDC1 than that of LoVo/mock. Blockade of syndecan-1 using small-interfering RNA (siRNA) gene knockdown canceled sensitization of LoVo/SDC1 to HB-EGF and diminished LoVo/SDC1 cell growth stimulated by HB-EGF. These results suggest that syndecan-1 overexpression may contribute to heparan sulfate binding growth factors sensitization and their promoting effects to cancer growth. Finally, we examined the effects of syndecan-1 on chemotherapy in the presence of heparan sulfate binding growth factors. In comparison to LoVo/mock, LoVo/SDC1 was resistant to some cytotoxic drugs (paclitaxel, 5-FU, or CPT-11) in the presence of HB-EGF or HGF. Blockade of syndecan-1 using siRNA diminished chemotherapy resistance of LoVo/SDC1 caused by HB-EGF. These findings demonstrate that syndecan-1 confers chemotherapy resistance in colon cancer through concomitant heparan sulfate binding growth factors sensitization. In summary, our results identify syndecan-1 as a functional coreceptor for heparan sulfate binding growth factors that activate their receptors and promote their downstream pathway in colon cancer cells. Syndecan-1 overexpression may contribute to tumor growth and the chemotherapy resistance characteristic of colon cancer. Therefore, therapeutic invention aimed at inhibiting this molecule may be useful in cancer treatment.
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 4108.
Abstract 4111: Induction of CXCL8 by TNFalpha and LIF (Leukemia Inhibitory Factor) in pancreatic carcinoma cells: Impact of CXCL8 as an autocrine growth factor
Authors: Kamohara Hidenobu , Hiyoshi Yoshiharu , Kurashige Junji , Kinoshita Koichi , Iwatsuki Masaaki , Baba Hideo 
Published: February 2011

Abstract
&lt;INTRODUCTION&gt;
Pancreatic carcinoma is one of the most lethal in the gastroenterogical malignant tumors. Chronic inflammation cause to cancer development and progression. CXCL8 is a CXC chemokine, which play an important role in neutrophil chemotaxis and activation. We previously reported that CXCL8 was produced by a variety of human carcinoma cells and tissues and that CXCL8 also promoted proliferation in pancreatic carcinoma cells. In the present study, we analyzed whether various cytokines affect cell proliferation by CXCL8 induction in pancreatic carcinoma cells.
&lt;METHODS and RESULTS&gt;
Eight examined pancreas carcinoma cell lines expressed CXCL8, CXCR1 CXCR2 and TNFRII mRNA constitutively in RPMI1640 medium without FBS. TNFalpha, LIF, IL1beta, IL6, CXCL8, or IFNbeta enhanced the expression of CXCL8 mRNA, but IL10 did not in Hs-700T cells. Especially, TNFalpha was a robust inducer. Actinomycin D suppressed and cycloheximide augmented CXCL8 mRNA which was induced by TNFalpha, LIF or not. The half-life of CXCL8 mRNA was 36.5 min by TNFalpha and 35.2 min without stimulation. That indicated the stability of CXCL8 mRNA was not affected by the stimulation of TNFalpha. Nextly, we examined the effect of CXCL8 on cell growth in Hs700T cells. Addition of recombinant TNFalpha or CXCL8 did not promoted cell growth of Hs700T. High amount of CXCL8 induced by TNFalpha or CXCL8 did not reponse to cell growth. On the other hand, LIF promoted cell growth after activation of STAT3 phosphrylation in Hs-700T cells by IP-Western blotting. The cell growth mechanism by LIF would depend on STAT3 activation. Anti-CXCL8 IgG significantly suppressed cell growth Hs-700T cells. Optimized concentration of CXCL8 would act as an autocrine growth factor in Hs-700T cells, which expressed CXCL8 mRNA highly without stimulation. Curcumin (Diferuloylmethane), NF-kappaB inhibitor, suppressed cell proliferation in Hs-700T cells.
&lt;CONCLUSION&gt;
These results suggest that CXCL8 play a pivotal role in development and progression of pancreatic cancer, and its expression is regulated by inflammatory cytokines. High concentration of CXCL8 produced by cancer cells could affect neigboring cells in pancreatic tumor microenvironment.
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 4111.