bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2644: Advanced nanoparticles with anticancer and MRI function for liver cancer in rats and rabbits
Authors: Hong Soon-Sun , Maeng Jin-Hee , Lee Ju-Hee , Choi Myung-Joo , Lee Hyunseung , Jung Kyung Hee , Lee Jeong-Sang
Published: February 2011

Recently drug delivery systems are focusing on the improvement of efficacy, the reduction of side effect, and diagnostics. In the current research, we developed a novel polymeric nanoparticle (YCC) with anticancer effect as well as magnetic resonance (MR) imaging, which encapsulate doxorubicin and superparamagnetic iron oxide (Fe3O4) with the size of 86.4 nm. The efficacy of the nanoparticles was evaluated in hepatocellular carcinoma (HCC) rats induced by the treatment of diethylnitrosamine for 13 weeks as well as in rabbit model with VX2 carcinoma in the liver, comparing with free doxorubicin (FD) and a commercial liposome drug DOXIL®. Each formula was administered intravenously at a dose of 2 mg/kg as doxorubicin three times with 5 day intervals, and anticancer efficacy of YCC were confirmed by MRI (T2) and hematoxylin-eosin (H&E) staining. The tumor size using MRI and the change of body weight were monitored for 10 days after the last administration of each formula. On the end of the study (Day 20th), all the formulas containing doxorubicin suppressed the tumor growth compared with the saline-treated group (control). Especially the relative tumor volume of YCC group was decreased three and four fold compared with the FD and DOXIL® group, respectively. Nevertheless, the loss of body weight as a side effect in YCC group was lower than FD or DOXIL® group. The results from Perls staining and transmission electron microscopy (TEM) confirmed that YCC was distributed in the liver, especially in tumor region rather than non-tumor region. In rabbit model with VX2 carcinoma in the liver, the relative tumor volume in YCC group was decreased two and four fold compared with the FD and DOXIL® group, respectively, as a result of MRI and tissue staining. Like in rat model, the loss of body weight as a side effect in YCC group was lower than FD or DOXIL® group. In both rat and rabbit model, YCC showed the MRI sensitivity comparable to a conventional MRI contrast agent (Resovist®) even in its lower iron content. In the immunohistochemical analysis, YCC group showed the lower expression of CD34 and Ki-67, markers of angiogenesis and cell proliferation, respectively, while apoptotic cells were significantly rich in the YCC group in terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Taken together, a novel nanoparticle YCC had anticancer efficacy better than conventional doxorubicin formulas (free doxorubicin and DOXIL®) with the function of MR imaging in rat and rabbit model with carcinoma in the liver. It was considered to a good candidate for the treatment of liver cancer monitoring the progress of the cancer using MRI.

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 2644.
Abstract 2658: Anti-GD3 antibodies are targeting molecules for delivery of RNAi to melanoma
Authors: Michael Wu W. , Pare Genevieve , Tron Victor A.
Published: February 2011

Passive non-targeted delivery of RNAi therapy into tumours has been shown to be neither very effective, nor selective in vitro and in vivo. Consequently in this study, siRNA will be linked to a drug delivery system that will express a ligand that specifically targets melanoma cells. It has been previously shown that melanoma over-expresses the cell surface ganglioside GD3, thus, the anti-GD3 R24 monoclonal antibody can function as a targeting molecule. To do so, a targeting antibody was conjugated to a coated cationic liposome (CCL) exterior to effectively deliver RNAi to melanoma. Cell surface expression of GD3 on Malme 3M, LOX IMVI and SK-MEL-5 cells was verified by flow cytometry. Internalized levels of siRNA within CCLs were the quantified via spectrometry. Encapsulation efficiency of siRNA was calculated to be greater than 70%. The quantification of antibody expression on CCLs was measured, illustrating that while non-targeted CCLs showed insignificant values of antibody as expected, antibody-conjugated CCLs presented a decreasing trend of expression that can be explained by the multi-step purification process for the final CCL products. Fluorescent microscopy results, from the combination of internalized siRNA and Tex615 RNA within CCLs, displayed greater fluorescence with antibody-conjugated CCLs than non-targeted CCLs when Malme 3M, LOX IMVI, and SK-MEL-5 cells were treated. Taken together, these results conclude that the use of GD3-targeted liposomes can promote siRNA delivery to melanoma cells.

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 2658.
Abstract 2673: In silico design of E2F1 mimics as potential anticancer agents
Authors: Kerrigan John E. , Abali Emine E. , Banerjee Debabrata , Hsieh Yi-Ching , Xie Xiaoqi , Bertino Joseph R.
Published: February 2011

Introduction. We report the construction of a homology model of the human E2F1-DP1 transcription factor complex with DNA to be used as a tool for design of drugs targeted to the promoter of this oncogene based on interference of transcription factor binding. Using shape-based docking and force field calculations, we investigate the development of novel penetratin-based (PED) peptides, obtained by phage display, (E2F1 mimics) as potential binders to the major groove of GC-rich regions of DNA.

Computational Procedures. The homology model of E2F1-DP1 complex with DNA was built using the Modeller (9v5) program. We used the DNA binding domains of the E2F1 and DP1 sequences (NCBI accession #'s AAC50719 and NP009042 respectively), and the x-ray crystal structure of the E2F4-DP2 complex with DNA (1CF7.pdb) as the primary template. The DNA from the crystal structure was modeled in place using the default spatial constraints in Modeller.

The PED based peptide models were built using the x-ray crystal structure of the antennapedia homeodomain-DNA complex (9ANT.pdb) as a template. The alignment is shown below (The penetratin portion is shown in bold and the variable portion is in italics.).

_aln.pos 10 20 30 40 50 60

9antA RQTYTRYQTLELEKEFHFNRYLTRRRRIEIAHALSLTERQIKIWFQNRRMKWKK—–EN

brt1 ————————————–RQIKIWFQNRRMKWKKHHHRLSH

_consrvd ****************

The PatchDock program was used to dock the peptides to the DNA structure from 1CF7.pdb based on shape complementarity. The best shape scoring major-groove binding orientation was selected and refined via energy minimization using the Amber ff99SB force field. Interaction energy scores were computed from the non-bonded energy terms of the Amber force field (Eint = ∑ Evdw + ∑ Eelec).

Data Summary. The HHHRLSH sequence is predicted to form a more stable complex compared to the GGGALSA sequence. The PED-HHHRLSH peptide was shown to have potent activity against several cancer cell lines (Xie, et al. AACR 2009). In the case of the HHHRLSH sequence, the model predicts enhanced stability in going from L-Arg (−646.0 kcal/mol) to D-Arg (−688.0 kcal/mol).

Conclusions. The α3 helix of E2F1 composed of KRRIYDITNVLEGI binds to the major groove of GC-rich DNA. The PED-HHHRLSH peptide complex with DNA (−646.0 kcal/mol) has similar interaction energy to that of the E2F1-DNA complex (−653.0 kcal/mol), is a stable α-helical structure, and is a potential E2F1 mimic.

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 2673.
Abstract 2680A: Small molecule induces apoptosis through the inhibition of signal transducers and activators of transcription 3 in hepatocellular carcinoma cells
Authors: Chen Kuen-Feng , Chen Wei-Lin , Tai Wei-Tien , Cheng Ann-Lii , Chen Pei-Jer , Shiau Chung-Wai
Published: February 2011

Sorafenib, a multiple kinase inhibitors, shows therapeutic effects in renal carcinoma (RCC) and hepatocellular carcinoma (HCC). Our data shows that the inhibition of signal transducers and activators of transcription 3, a transcription factor involving in cell proliferation and survival, represents a major mechanism whereby sorafenib induces apoptosis in HCC cells. Sorafenib inhibits the phosphorylation of STAT3 and subsequently reduced the STAT3-regulating protein, Mcl-1, survivin and cyclinD1. In addition, the activity of SHP-1, a protein tyrosine phosphatase, is increased in HCC cells by the treatment of sorafenib. We hypothesize that the antitumor effect of sorafenib can be attributed to the level STAT3 and the activity of SHP-1. Therefore, we design and synthesize a sorafenib derivative, SC-1, which is devoid of kinase activity. SC-1 shows higher potency in inducing apoptosis than sorafenib. Therefore, SC-1 provides a proof of concept for the design of a new class of agents that enhance SHP-1 activity along and synergize with therapeutic agent in HCC.

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 2680A.
Abstract 2691: Prognostic significance of heat shock protein 90 expression in patients with advanced gastric cancer
Authors: Dote Hideaki , Nishimura Rieko , Yamamoto Tamami , Iguchi Haruo , Nozaki Isao , Hato Shinji , Kobatake Takaya , Ohta Kouzi , Aogi Kenjiro , Kubo Yoshiro , Tanada Minoru , Kurita Akira
Published: February 2011

Purpose: Heat shock protein 90 (HSP90) is a chaperone mediating the folding and stabilization of many oncoproteins. Considerable attention has been focused on the role of the HSP90 in the therapeutic strategy of molecular targeting HSP90. Recently it has reported that Trastuzumab, a recombinant monoclonal antibody against HER2, plus chemotherapy improved survival in HER2-positive gastric cancer patients. This study was designed to delineate the clinical implications of Hsp90 and HER2 immunoexpression in advanced gastric cancer.

Material and Methods: The study group comprised 47 patients who underwent gastrectomy at Shikoku Cancer Center Hospital excluding patients with stage I. According to the TNM classification, 18 tumors were identified as being stage II, 25 stage III, 4 stage IV. Using immunohistochemical techniques, we analyzed the expressions of HSP90 and HER2 on formalin-fixed paraffin-embedded specimens of surgically removed primary tumors. Immunostaining was graded as follows: low-HSP90 (defined as weaker staining compared with adjacent normal gastric mucosa), moderate-HSP90 (defined as equal), high-HSP90 (defined as stronger). HER2 expression was graded using a 4-point scale according to the criteria of HER2 membranous staining, which have been widely accepted. Chi-square test, Kaplan-Meier and Cox regression analysis were used for statistical analysis.

Results: Low-HSP90 expression was found in 13 tumors (28%) and HER2 high-expression (moderate to strong membrane staining in > 10% of tumor cells) in 13 tumors (28%). Low-HSP90 was significantly detected in diffuse type (p = 0.0047) and was associated with clinicopathological parameters involved with tumor progression, including the depth of tumor invasion (p = 0.049) and advanced stage tumor (p = 0.029). Of note, High-HSP90 and HER2 overexpression (3+) were mutually exclusive (P=0.031). Kaplan-Meier survival analysis determined that tumors with low-HSP90 expression were statistically associated with worse disease-free survival (p = 0.036), and tended to detect poor postoperative survival (p = 0.084). Multivariate survival analysis showed that HSP90-negative expression [hazard ratio (HR) 3.05, 95% confidence interval (95% CI) 1.02-9.18; p = 0.046] and lymph node metastasis (HR 5.55; 95% CI 1.68-18.2; p = 0.005) were significant predictors of poor postoperative survival.

Conclusions: Our results indicate that low-HSP90 expression correlated with more aggressive disease and poor prognosis, and that HSP90 and HER2 overexpression might separately be molecular targets in gastric 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 2691.
Abstract 2795: Serum IGF-I, IGF-II, IGFBP-3, and IGF-I/IGFBP-3 molar ratio and risk of pancreatic cancer in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial
Authors: Douglas Jason B. , Silverman Debra T. , Pollak Michael N. , Tao Yuzhen , Soliman Amr S. , Stolzenberg-Solomon Rachael Z.
Published: February 2011

Experimental evidence suggests that an overexpression of insulin-like growth factor-I (IGF-I) is implicated in human pancreatic tumors. Moreover, increased IGF-II and decreased insulin-like growth factor binding protein-3 (IGFBP-3) concentrations are found in a number of cancers. We conducted a nested case-control study in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial cohort of men and women 55-74 years of age at baseline, to test whether pre-diagnostic circulating IGF-I, IGF-II, IGFBP-3, and IGF-I/IGFBP-3 molar ratio concentrations were associated with exocrine pancreatic cancer risk. Between 1994 and 2006, 187 incident cases of pancreatic adenocarcinoma occurred (follow-up to 11.7 years). Two controls (n=374),who were alive at the time the case was diagnosed, were selected for each case and matched by age, race, sex and date of blood draw. We calculated odds ratios (ORs) and 95% confidence intervals (CIs) using conditional logistic regression, adjusting for smoking. In the multivariable adjusted model, IGF-I, IGF-II, and IGFBP-3 concentrations were not associated with pancreatic cancer (highest compared with lowest quintile, OR=1.23, 95% CI 0.67-2.25, p-trend=0.12; OR=0.98, 95% CI 0.52-1.85, p-trend=0.52; and OR=1.15, 95% CI 0.62-2.16, p-trend=0.90). However, a significant positive trend was observed with high IGF-I/IGFBP-3 molar ratio levels in the smoking-adjusted model (OR=1.46, 95% CI 0.78-2.74, p-trend=0.04). A higher IGF-I/IGFBP-3 molar ratio may represent increased free IGF-I and be a risk factor for pancreatic 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 2795.
Abstract 2927: Gefitinib, an EGFR inhibitor, prevents progression of PanINs to pancreatic carcinoma in a preclinical conditional LSL-K-rasG12D transgenic mice model
Authors: Mohammed Altaf , Janakiram Naveena B. , Qian Li , Madka Venkateshwar , Ely Misty , Lightfoot Stan , Steele Vernon E. , Rao Chinthalapally V.
Published: February 2011

Pancreatic ductal adenocarcinoma (PDAC), the most common pancreatic malignancy is a devastating disease with dismal prognosis (5-year survival rate of less than 5%), and is the fourth leading cause of cancer deaths in the USA. Developing novel strategies to prevent or delay the progression of pancreatic cancer is currently of intense interest. We studied chemopreventive efficacy of gefitinib, an EGFR inhibitor on pancreatic intraepithelial neoplasms (PanINs) and adenocarcinoma in conditional LSL-K-RasG12D transgenic mice. LSL-K-RasG12D and P48Cre mice were bred and offspring of activated K-rasG12D were generated. Six-week old male KrasG12D (20/group) and wild-type (12/group) mice were fed (AIN-76A) diets containing 0, 100, and 200 ppm gefitinib for 35 weeks. At the time of sacrifice, pancreas were collected, weighed, and were evaluated histopathologically for PanINs and PDAC. To understand the molecular mechanisms, expression levels of PCNA, p38, cyclin D1, C2GNT, RhoA, β catenin, pERK, EGFR and Cav1, were determined in pancreatic tissues by IHC, immuno-fluorescence, immunoblotting and/or RT-PCR. Results suggest that mice fed with 100 and 200 ppm gefitinib had significantly reduced (p<0.0001) ductal adenocarcinomas incidence than control diet fed mice. Importantly, significant reduction of pancreatic ducts with carcinoma (>94%, p< 0.0001) and suppression of PanIN 3 (∼34%) was observed in mice fed diet with gefitinib. We did find statistically significant dose dependent inhibition of PanIN 1 & 2 lesions between control and gefitinib fed groups. Also, mice fed gefitinib showed reduced pancreatic tumor weights (∼50%) compared to control diet fed mice. Furthermore, treated mice showed >75% of pancreas free from carcinoma. Also, pancreas of mice fed gefitinib diets showed significantly reduced expressions of PCNA, p38, EGFR, RhoA, C2GNT, β-catenin, pERK, caveolin-1, and a significant increase in cyclin B1 (p<0.005) levels compared to pancreas of control diet fed mice. In summary, our results suggest that gefitinib prevents the progression of pancreatic cancer precursor lesions in PanIN 1, 2 and 3 stages from further progressing to adenocarcinoma in a preclinical model. These data highlight the promise of chemoprevention in pancreatic cancer and administration of EGFR inhibitors represents an important strategy to prevent pancreatic cancer in high-risk patients.

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 2927.
Abstract 3001: MicroRNAs regulating wnt signaling are downregulated in malignant pleural mesothelioma
Authors: Gee Gretchen V. , Koestler Devin C. , Christensen Brock C. , Sugarbaker David J. , Ugolini Donatella , Ivaldi Giovanni P. , Resnick Murray B. , Houseman E. Andreas , Kelsey Karl T. , Marsit Carmen J.
Published: February 2011

Malignant pleural mesothelioma is a rapidly fatal disease whose diagnosis, particularly through less invasive techniques such as analysis of pleural effusion, can be challenging. Currently, a commercially available diagnostic test based on microRNA expression patterns is purported to distinguish between mesothelioma and lung adenocarcinoma. Yet, the biological basis of this technology has not been reported in the literature, and little research has been aimed at determining how differential miRNA expression contributes to the differences in pathogenesis between these diseases, both of which can be caused by asbestos exposure. We sought to illuminate the molecular differences between mesothelioma and lung adenocarcinoma by using microRNA microarrays to identify patterns in the most differentially expressed microRNAs. From this, we identified a panel of microRNAs, including members of the miR-200 gene family, that were all downregulated in malignant pleural mesothelioma compared to lung adenocarcinoma. Using the more sensitive detection method of quantitative RT-PCR on an independent series of tumors, we validated the specificity of these alterations in 100 malignant pleural mesotheliomas and 32 lung adenocarcinomas. Further examination of the relationship between these microRNAs revealed that they are predicted to act as redundant regulators of wnt signaling, and suggests a role for this pathway in tumor progression. This research points to novel approaches utilizing miRNAs whose decreased expression is unique to mesothelioma as potentially suitable for rapid diagnosis, and reveals important new targets for the treatment of this deadly disease.

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 3001.
Abstract 3030: An exosome isolation system for serum-based cancer biomarker discovery
Authors: Wu Fangting , Antes Travis J.
Published: February 2011

Exosomes are 40-100 nm membrane vesicles secreted by most cell types, including all cancer cells. While the exact biological functions of exosomes remain unclear, exosomes can mediate communication between cells, facilitating processes such as antigen presentation and in trans signaling to neighboring cells. The abundance of exosomes present in the circulation of cancer patients increases with tumor grade. Studies identifying protein and microRNA biomarkers in these circulating exosomes can reveal the tumor type and origin. These recent studies highlight the growing use and need of standardized exosome enrichment. What is missing in this research area is a method for simple, rapid and reproducible exosome purification to facilitate research into the role of exosome in both physiological and pathological process, as well as possible diagnostic and therapeutic potentials. In this study, SBI utilizes proprietary polymer mixture (ExoQuick) to precipitate exosome from human serum samples. Using ExoQuick, we successfully observe small RNAs in the exosome pellets and detect microRNAs by qPCR. Exosome precipitation from serum samples was verified using Western blot analysis on the ExoQuick supernatant and pellet fractions and probed with antibodies against two commonly found exosome biomarker proteins, CD63 and CD9. Robust signals for CD63 and for CD9 were observed only in the exosome pellet fractions but not in the serum supernatant. These preliminary data provide proof-of-concept for our approach in using polymer-based, one-step exosome purification method from human serum samples. The technologies developed in this study will advance the exosome research field by providing a simple and standardized exosome isolation method that will allow high-throughput exosome analysis to enable major strides forward in serum-based microRNA biomarker discoveries for cancer diagnostics and prognostics tools.

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 3030.
Abstract 3051: mRNA and microRNA expression profiles integrated with drug sensitivities of the NCI-60 human cancer cell lines
Authors: Liu Hongfang , D'Andrade Petula , Fulmer-Smentek Stephanie , Lorenzi Philip , Kohn Kurt , Weinstein John , Pommier Yves G. , Reinhold William C.
Published: February 2011

We present here new NCI-60 profiling studies of mRNA and miRNA expression using the 41,000-probe Agilent Whole Human Genome Oligo Microarray and the 15,000-feature Agilent Human miRNA Microarray V2. Expression levels were determined for ∼21,000 genes and 723 human miRNAs. The resulting data sets are freely available and searchable online at http://discover.nci.nih.gov in our CellMiner relational database package. The profiling included technical replicates, with six and eight cell lines assayed in quadruplicate for mRNA and miRNA, respectively. The remaining cell lines were assayed in duplicate. Our analysis indicates high reproducibility for both platforms and an essential biological similarity across the various cell types. The expression levels were integrated with our previously published 1,429-compound database of anticancer activity in the NCI screen. Large blocks of both mRNAs (∼2000) and miRNAs (∼200) with unidirectional correlation to ∼1300 drugs including 121 drugs with known mechanisms of action were identified. The data sets presented here will facilitate recognition of the groups of mRNAs, miRNAs and drugs that potentially affect and interact with one another.

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 3051.
Abstract 3095: JMJDX interacts with DEAH (Asp-Glu-Ala-His) box polypeptide 9 (DHX9)
Authors: Unoki Motoko , Nakamura Yusuke , Hamamoto Ryuji
Published: February 2011

We identified JMJDX as a candidate oncogene. Expression of JMJDX was up-regulated in various cancers including bladder cancer. Knockdown of JMJDX suppressed cell growth, suggesting that the protein may be involved in cancer cell growth. The protein possesses a JmjC domain. The domain is often found in lysine demethylases. To identify substrates of JMJDX, we performed immunoprecipitation and subsequent mass spectrometric analysis. In the result, we found that JMJDX physically associated with DHX9, which is also known as RNA helicase A. DHX9 is a DNA helicase and also works as an RNA helicase. DHX9 has a nuclear transport domain (NTD), which is known to necessary and sufficient for its bi-directional nuclear trafficking. It was reported that arginine methylation at the NTD domain is required for the NTD-mediated nuclear import. Thus, JMJDX may demethylate the methylated arginines at the domain and control its transportation between nucleus and cytoplasm. If our hypothesis is true, JMJDX will be the first arginine demethylase, because JMJD6, which was reported as the first arginine demethylase, was recently turned out that it is actually a lysine hydroxylase. DHX9 is thought to have oncogenic functions in cancer cells. In addition, expression levels of JMJDX are very low in normal tissues. Therefore, we think that JMJDX can be an anti-drug target with less adverse reactions.

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 3095.
Abstract 3124: Platelet-derived growth factor receptor α cooperates with loss of p16/INK4a and p19/ARF to enhance tumorigenesis through the PI3K- and SHP-2-mediated signaling in the brain
Authors: Liu Kunwei , Hu Bo , Feng Haizhong , Kazlauskas Andrius , Smith Erin , Symed Karen , Bachoo Robert , Cheng Shi-Yuan
Published: February 2011

Glioblastomas are the most common and lethal cancer in the central nervous system. Comprehensive genetic studies showed that in high-grade gliomas, overexpression of platelet-derived growth factor receptor alpha (PDGFRα) ranks third among top amplified genes and frequently co-occurs with loss of tumor suppressor genes, p16/INK4a and p14/ARF (p19/ARF in mice), in these high-grade glioblastomas. However, the biological consequences of PDGFRα overexpression in INK4a/ARF-deficient glial cells and glioma cells remain elusive. Here we show that activation of PDGFRα signaling by overexpressing PDGFRα and/or PDGF-A transformed Ink4a/Arf-deficient mouse astrocytes in vitro and rendered these cells highly tumorigenic in the brain of mice. Consistently, in INK4a/ARF null but not wild-type human glioma cell lines that express endogenous PDGFRα, overexpression of PDGF-A significantly enhances glioma formation in the brain. Re-introduction of p16/INK4a into Ink4a/Arf-null astrocytes suppressed PDGFRα-induced cell transformation in vitro and glioma formation in the brain. Conversely, cellular depletion of endogenous p16/INK4a by RNA interference in human glioma cells markedly enhances colony formation in vitro when compared to the controls. Mechanistically, abrogation of intrinsic tyrosine kinase activity of PDGFRα or interruptions of signaling modules at PDGFRα (Y to F mutations at specific tyrosine residues) that lost binding capacity for PI3K or Shp-2, but not PLC gamma or c-Src, significantly diminished PDGFRα-induced glioma formation in the brain. In vitro, specific YF mutations or inhibitors of PI3K or Shp-2 attenuated Akt activation and PDGFRα-induced colony formation. Furthermore, inhibition of Shp-2 by its inhibitors and siRNAs disrupts the interaction of PI3K with PDGFRα that is critical for gliomagenesis of Ink4a/Arf null cells. In summary, our data suggests that activation of PDGFRα signaling, particularly through the PI3K and Shp-2 pathways, in INK4a/ARF deficient astrocytes and glioma cells induce gliomagenesis in the brain. Our findings provide critical insights into interactions of genetic lesions frequently found in clinical glioma specimens and will be useful for identification of new therapeutic targets for treatments of patients with malignant gliomas.

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 3124.
Abstract 3130: Interacting protein of granulin-epithelin precursor in liver cancer
Authors: Lam C.Y. , Poon T.c.w. , Cheng K.C. , Cheung S.T.
Published: February 2011

Background and Objective: Granulin-epithelin precursor (GEP), a novel growth factor, has been identified as a potential therapeutic target from our earlier genome-wide expression profile study on liver cancer. GEP over-expressed in liver cancer, and regulated proliferation, invasion and tumorigenesis. We demonstrated the therapeutic approach of GEP targeted therapy in human cancer using the home-made GEP monoclonal antibody on animal model, and GEP antibody showed dose-dependent inhibitory effect on tumor growth. However, the exact mechanism on how GEP executes its biological function, and in particular its interacting protein partners, is largely unknown. The present study aims to identify and characterize the GEP interacting protein in liver cancer.

Methods: Coimmunoprecipitation (co-IP) was employed to investigate the protein-protein interacting partners of GEP. Total protein was extracted from liver cancer cells Hep3B, and specific anti-GEP monoclonal antibody was used to precipitate its interacting protein partner. The precipitated proteins were analyzed by SDS-PAGE. The extra band (compared with control 1: lysate alone, and control 2: antibody alone) was excised and analysed for protein identification using mass-spectrometry. The interaction of the GEP partner was validated in additional liver cancer cells Hep3B and HepG2 with swap co-IP and western blot using specific antibodies. The expression of GEP and the interacting partner in liver cancer cells and clinical samples were further examined by immunohistochemistry, western blot and real-time quantitative RT-PCR.

Results: The GEP co-IP analysis showed an extra band at 30kDa, and the protein was identified as tropomyosin 3 (TPM3). The interaction of GEP and TPM3 was demonstrated in additional liver cancer cells Hep3B and HepG2, with co-IP using specific TPM3 antibody and detection by western blot using GEP monoclonal antibody. Elevated TPM3, both on mRNA and protein levels, were observed in the liver cancer tissues compared to cirrhosis, hepatitis and normal livers. Overexpression of TPM3 was demonstrated in 74% of liver cancer tissues compared to their paralleled adjacent non-tumor livers. Immunohistochemistry demonstrated cytoplasmic TPM3 protein, and co-localized with GEP protein. High TPM3 level was associated with advanced tumor stage.

Summary: TPM3 locates at 1q21.2, a region with recurrent chromosomal gain in liver cancer. The TPM3-ALK fusion gene has been reported to associate with transformation, proliferation, invasion and metastasis. The current study demonstrated that TPM3 is an interacting partner of GEP in liver cancer, and strong TPM3 was associated with advanced stage. Thus, TPM3/GEP signaling should play important role in liver 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 3130.
Abstract 3198: Growth factors protect intestinal stem cells from radiation-induced apoptosis by suppressing PUMA through the PI3K/AKT/p53 axis
Authors: Qiu Wei , Leibowitz Brian , Zhang Lin , Yu Jian
Published: February 2011

Gastrointestinal toxicity is the primary limiting factor in abdominal and pelvic radiotherapy, but has no effective treatment currently. We recently showed a critical role of the BH3-only protein PUMA in acute radiation-induced GI damage and GI syndrome in mice. Growth factors such as IGF-I and bFGF have been shown to protect against radiation-induced intestinal injury, though the underlying mechanisms remain to be identified. We report here the suppression of PUMA through the PI3K/AKT/p53 axis in the intestinal stem cells (ISCs) as a novel molecular mechanism of growth factor-mediated intestinal radioprotection. IGF-I or bFGF impaired radiation-induced apoptosis, and the expression of PUMA and p53 in the crypt cells and intestinal stem cells. Using colonic epithelial cells that undergo PUMA-dependent and radiation-induced apoptosis, we found that a PI3K inhibitor, dominant-negative PI3K, or Mdm2 antagonist restored the induction of PUMA, p53 and apoptosis in the presence of growth factors. In contrast, overexpression of AKT suppressed the induction of PUMA and p53 by radiation. Furthermore, inhibiting PI3K or activating p53 abrogated growth factor-mediated suppression of apoptosis and PUMA expression in the intestinal crypts and stem cells following radiation.

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 3198.
Abstract 3212: Development of a chemiluminescence-based single telomere length analysis (STELA) method
Authors: Heaphy Christopher M. , Haffner Michael C. , Meeker Alan K.
Published: February 2011

Carcinogenesis is a multistep process characterized by the accumulation of genetic alterations that drive the transformation of normal cells to malignant cells. Telomeres are specialized nucleoprotein structures that function to protect and stabilize the ends of eukaryotic chromosomes by preventing chromosome fusions, masking double strand DNA break damage signals and preventing exonucleolytic chromosomal degradation. It is well established that telomere shortening is present in precursor and invasive stages of many carcinomas. Additionally, numerous investigations have demonstrated that altered telomere length predisposes cells to possess the properties necessary for metastasis formation, and therefore represents a predictor of clinical outcome. However, most of these studies have used assays that measure the average telomere length of all the cells in a particular population. To circumvent this problem, Baird et al. (Nature Genetics, 2003) previously described a PCR-based single telomere length analysis (STELA) method that could accurately measure the full spectrum of telomere lengths from a specific chromosomal arm. Although this method represents a substantial improvement over existing methods, the assay requires the use of radioactive material. Here, we describe a modification of the STELA assay which uses chemiluminescence instead of radioisotopes for product visualization, while maintaining the overall sensitivity of the detection of a wide range of telomere lengths. This new methodology has tremendous potential to illuminate the mechanisms that support the initiation of cancer and may provide an avenue for investigators to address the full extent and significance of telomere shortening in the tumorigenic process at the single chromosome level. The modified assay described here will allow for more widespread use of this technique among cancer researchers.

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 3212.
Abstract 3216: CREG1, an epigenetically regulated gene, cooperates with p16INK4a in cellular senescence
Authors: Moolmuang Benchamart , Tainsky Michael A.
Published: February 2011

By passing cellular senescence, an irreversible growth arrest of cells that is activated in normal cells upon shortening of telomere and other cellular stress, to become immortal is one of the prerequisites for carcinogenesis. Using spontaneously immortal Li-Fraumeni Syndrome (LFS) fibroblasts, we found that CREG1 (Cellular Repressor of E1A-stimulated Genes1) is one of the genes whose expression fit the criteria of senescence-associated genes, decrease expression during immortalization and increase in senescence. Moreover, we found that epigenetic mechanism regulates CREG1 expression. CREG1 is a secreted glycoprotein that was shown to bind RB-family pocket proteins and inhibits E2F transactivation activity. Therefore, we hypothesize that CREG1 plays a role in cellular senescence involving p16INK4a/Rb pathway.

Ectopic expression of CREG1 in the immortal LFS cell lines decreases cell proliferation but does not directly induce senescence. We confirmed this in osteosarcoma and fibrosarcoma cancer cell lines commonly seen in Li-Fraumeni Syndrome. Because CREG1 was shown to interact with RB pocket proteins in vitro, we tested that growth suppression by CREG1 may depend on the phosphorylation status of Rb. We found that p16INK4a is also downregulated in immortal cells and that coexpression of CREG1 and p16INK4a, and inhibitor of CDK and Rb phosphorylation, has a greater effect than CREG1 and p16INK4a alone to reduce cell growth, induce cell cycle arrest and cellular senescence in immortal LFS, osteosarcoma and fibrosarcoma cell lines. Moreover, cooperation of CREG1 and p16INK4a inhibits the expression of cyclin A and cyclin B by inhibiting promoter activity thereby decreasing mRNA and protein levels; these proteins are required for S-phase entry and G2/M transition. Whether CREG1 directly involved in transcriptional repression of cyclin A and B mediated by p16INK4a will be confirmed by ChIP assay. In conclusion, we are the first to find that CREG1 enhances p16INK4a-induced senescence by transcriptional repression of cell cycle mediated genes.

Analysis of how CREG1 and its cooperation with p16INK4a regulate cell cycle and cellular senescence may provide better understanding of cellular immortalization, an early step in human tumorigenesis.

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 3216.
Abstract 3228: Dual HER2 and EGFR inhibitor BIBW2992 have dramatic activity in a HER2 wild type/EGFR L858R murine lung cancer model
Authors: Li Danan , Chen Zhao , Chen Liang , Liang Mei-chih , Walton Zandra , Sanofsky Benjamin , Janne Pasi , Wong Kwok-Kin
Published: February 2011

Dual HER2 and EGFR inhibitor BIBW2992 have dramatic activity in a HER2 wild type/EGFR L858R murine lung cancer model

HER2 (ERBB2) is a proto-oncogene that belongs to EGFR tyrosine kinase receptor family. HER2 genomic locus amplification and HER2 protein over-expression detected by IHC have been identified in many human cancer types including breast, brain and lung cancer. Simultaneous over-expression of both HER2 and EGFR occurs in a subpopulation of lung cancer patients. Here we developed a transgenic mouse model with inducible wild type HER2 expression in type II pneumocyte. Continual HER2 over-expression in the lung epithelial compartment leads to peripheral lung adenocarcinoma that responds dramatically to targeted anti-HER2 therapeutics. Mice over-expressing both EGFR L858R and the wild type HER2 alleles do not significant accelerate tumorigenesis or alter of tumor histology as compared to tumors generated by the wild type HER2 single alleles. The HER2/EGFR driven murine lung tumors showed mild and transient response to the EGFR inhibitor, gefitinib. This result mimics the observation from clinical patients. However, the dual HER2/EGFR inhibitor, BIBW2992, is much more effective against these HER2/EGFR driven lung cancers as measured by tumor regression and over all survival. Our data suggested that targeting therapy against both EGFR and HER2 is a better strategy in lung cancer patients with co-expression of EGFR and HER2 when compared with treatment against EGFR alone. Clinical trial for dual HER2/EGFR inhibitors in this population should be considered.

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 3228.
Abstract 3266: Alpha2 and beta-adrenergic action on human breast cancer cells growing in nude mice
Authors: Piñero Cecilia Pérez , Entschladen Frank , Lüthy Isabel Alicia
Published: February 2011

Breast cancer, the most common cancer amid women in the majority of countries, is among the most stressful of diseases. The principal effectors of the stress system include the catecholamines epinephrine and norepinephrine. The catecholamines bind to α1-, α2- and β-adrenoceptors (AR). Our group has recently described α2-adrenoceptors in human tumor and non-tumor breast cell lines by reverse transcription (RT)-PCR, immunocytochemistry and binding assays. Moreover, the stimulation by α2-adrenoceptor agonists was associated with increased cell proliferation and tumor growth. On the other hand, we observed a reduction in tumor growth when the animals are treated with the pharmacological α2-antagonist RAUWOLSCINE (RAW). The expression of β2-AR has been described in different experimental models for breast cancer. The aims of the present work were: to assess the effect of β-adrenergic compounds in the human breast cancer cell lines growing in nude mice (IBH-4 and IBH-6); to study the migration potential of these cell lines using video microscopy in three dimensional collagen lattices knowing that migration of tumor cells is a prerequisite for invasion and metastasis development. The daily administration of 1,2mg/kg of the SALBUTAMOL (SALB) significantly diminished tumor growth. The same result was seen when using the α2-antagonist RAW (0,5mg/kg). As an example for tumor IBH-4 treated with SALB, on day 20: 444.16 ± 76.59mm3 vs. 843.40 ± 189.41 mm3, p<0.05; RAW on day 24: 775.04 ± 68.12mm3 vs 1237.84 ± 134.97mm3 p<0.01. Tumor IBH-6, on day 21: SALB: 125.40 ± 25.42mm3 vs. 579.40 ± 207.96mm3 p<0.01. The combined treatment of SALB + RAW did not exert any synergic or additive effect compared with each independent treatment. ERK phosphorylation is in agreement with tumor growth results. When analyzing the migration potential of the cells in response to adrenergic stimulation by three dimension video microscopy in collagen lattices, cell migration was decreased when the cells were treated with RAW, SALB and PROP and increased when treated with epinephrine. The migratory activity for each treatment was calculated as the portion of cells which was locomotory active (% locomoting cells). We conclude that both RAW, acting as α2-AR antagonist and SALB, acting through β-AR, inhibit tumor growth. Regarding cell migration, the results are promising and in agreement with the in vivo experiments. These compounds, devoid of serious side-effects, could eventually be employed to inhibit tumor growth. In the future, it might be useful to analyse the metastatic potential of the cells.

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 3266.
Abstract 3297: CD90/Thy-1-positive cells in the peripheral blood of tumor patients co-express CD44
Authors: Wagner Kathleen , Berndt Alexander , Hochhaus Andreas , Clement Joachim H.
Published: February 2011

Aims: Interactions between epithelial tumor cells and the surrounding milieu are an essential regulatory component of tumor development. Especially the contribution of the crosstalk between epithelial tumor cells and tumor-associated fibroblasts (TAFs) on tumor progression and metastasis formation is of emerging interest. Therefore, we ask whether circulating TAFs could be detected in the peripheral blood of tumor patients and how they could be further characterized.

Methods: CD90/Thy-1 is a putative marker of TAFs. A fluorescence-scanning-cytometer (scanR) was used to detect and quantify vital CD90-positive cells in blood samples from individual tumor patients (e.g. breast, bladder, kidney). For further analysis CD90-positive cells were separated from leukocyte fractions pooled from different tumor patients using an immunomagnetic cell separation technology (ROBOSEP®). The CD90-positive fraction was subsequently analyzed by immunofluorescence and immunohistochemistry. We focused on the expression of other stroma markers like FAP-alpha and cell surface proteins like CD44 which are involved in metastasis formation.

Results: In cell culture experiments we established CD90 as a highly specific marker for fibroblasts. The amount of CD90 positive cells in whole blood samples varied from 0 up to 54,000 cells/ml and changed over time. The CD90-positive cells were enriched immunomagnetically from the leukocyte fraction pooled from tumor patients. By immunofluorescence we approved the cell vitality and verified that the separated cells do not belong to the sub-population of CD34-positive blood stem cells. Blood samples from more than 300 patients with solid tumors were tested for the presence of CD90-positive cells. Immunofluorescence double staining of the separated cells showed the co-expression of CD90 and CD44, which participates in a wide variety of cellular functions like hematopoiesis and tumor metastasis formation. Further immunohistochemical analysis demonstrated the expression of CD29 which is involved in metastasis formation and CD105 a marker for activated fibroblasts in a subset of the CD90-positive cell fraction. Conclusions: We could show that CD90-positive cells circulate in the peripheral blood of tumor patients. The additional expression of other stroma and tumor associated proteins on the cells could argue for their involvement in aspects of tumor progression. Ongoing studies should evaluate their suitability as prognostic factor.

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 3297.
Abstract 3300: MYC regulation of a “poor prognosis” metastatic cancer cell state
Authors: Wolfer Anita , Wittner Ben S. , Irimia Daniel , Flavin Richard J. , Lupien Mathieu , Gunawardane Ruwanthi N. , Meyer Clifford A. , Lightcap Eric S. , Tamayo Pablo , Mesirov Jill P. , Liu Xiaole S. , Shioda Toshihiro , Toner Mehmet , Loda Massimo , Brown Myles , Brugge Joan S. , Ramaswamy Sridhar
Published: February 2011

Gene expression signatures are used clinically as prognostic tools to determine the risk of individual patients with localized breast tumors developing distant metastasis. However, we lack a clear understanding of whether these correlative biomarkers link to a common biological network that regulates metastasis. METHODS: In order to address this question, we analyzed thirteen different well-described (including two in clinical use) “poor outcome” cancer signatures in a cell line model of estrogen receptor positive breast cancer (MCF7) manipulated by diverse oncogenic stimuli. RESULTS: We find that the c-MYC oncoprotein coordinately regulates the expression of these thirteen different “poor outcome” cancer signatures. In addition, functional inactivation of MYC in human breast cancer cells specifically inhibits their distant metastasis in vivo and invasive behavior in vitro. CONCLUSION: These results suggest that MYC oncogene activity, as reflected by “poor prognosis” signature expression, may be necessary for the translocation of “poor outcome” human breast tumors to distant sites.

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 3300.