bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3078: CD44 mRNA stability is regulated by RNA binding protein HuR in ovarian cancer cells
Authors: Miletti-Gonzalez Karl E. , Hostetter Christine L. , Ravindranath Abhilash K. , Muthukumaran Neelakandan , Keen Judith C. , Rodriguez-Rodriguez Lorna
Published: June 2012

CD44, a major hyaluronic acid (HA) receptor, is involved in cell migration, metastasis and tumor progression. While in normal ovarian epithelium CD44 is not expressed, ovarian carcinoma cells express multiple CD44 isoforms as the tumor progresses and eventually CD44 expression is abrogated in advanced ovarian cancer metastatic sites. We as well as others have observed that the levels of CD44 mRNA transcript not always correspond to the CD44 protein expression levels. This observation suggests that CD44 is post-transcriptionally regulated. To test this hypothesis we first analyzed the CD44 3’ untranslated region (3’ UTR) sequence and found numerous AU-rich elements (ARE) in this region. Some of these sequences are known HuR binding site consensus sequences. HuR is an RNA binding protein that is known to stabilize transcripts. HuR is expressed in the nucleus and is transported to the cytoplasm where it functions as an mRNA stabilizer. We performed mRNA half-life experiments that showed a two-fold reduction in the stability of CD44 transcript in siHuR transfected ovarian cancer cells. Consistent with this result, treatment of these cells with inhibitors of DNA methylation and histone deacetylation (i.e., 5-azacytidine and Trichostatin A, respectively) known to diminish the presence of HuR in the cytoplasm, also decreased the stability of CD44 mRNA. Western blot analysis showed the expected reduction in cytoplasmic HuR as well as down-regulation in CD44 protein expression. A direct interaction of HuR with the CD44 3’ UTR was detected by biotin pulldown assays. These results indicate that HuR posttranscriptionally stabilizes the expression of CD44 in the cytoplasm.

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 3078. doi:10.1158/1538-7445.AM2011-3078
Abstract 3086: Interactions of nucleolin and ribosomal protein RPL26 in the translational control of human p53 mRNA
Authors: Chen Jing , Guo Kexiao , Kastan Michael B.
Published: June 2012

Our lab has previously shown that the ribosomal protein RPL26 enhances p53 translation and induction after DNA damage and that this regulation depends upon interactions between the 5′ and 3′ untranslated regions (UTR) of human p53 mRNA (Takagi et al Cell, 2005; Chen & Kastan, Genes & Dev, 2010). In contrast, nucleolin (NCL), a multifunctional nucleolar protein, suppresses the translation of human p53 mRNA and its induction after DNA damage. Interestingly, RPL26 and NCL also interact with each other. In order to understand how these two proteins cooperate with/antagonize each other to regulate p53 translation after stress, we first recapitulated the translational control of p53 by NCL in both an in vitro transcription/translation system and a cell-based, luciferase reporter system. In both systems, NCL repression required the presence of both the 5′UTR and 3′UTR of p53 mRNA. Further, NCL binds to the same 5′-3′ UTR interaction region that is critical for RPL26 recruitment after DNA damage. We also observed that NCL protein interacts with itself; the ability of NCL to dimerize/oligomerize and its reported ability to stabilize double strand nucleic acid structures suggests the possibility that NCL may stabilize the double-strand RNA structure involving the two p53 mRNA UTR's. Domain mapping demonstrated that the RNA binding domain of NCL participates in binding to p53 mRNA, is required for both NCL dimerization and NCL-mediated translational repression, and is the domain of NCL that interacts with RPL26. RPL26 was able to disrupt NCL dimerization in cell-based immunoprecipitation assays. We also identified the domain of RPL26 that is required to interact with nucleolin and found that point mutations in this region of RPL26 reduce RPL26-NCL interactions and attenuate both RPL26 binding to human p53 mRNA and p53 induction by RPL26. These observations taken together suggest a model for the interplay of NCL and RPL26 in the translational control of human p53 mRNA after DNA damage: NCL binds both p53 UTR's and NCL dimerization/oligomerization facilitates the double strand RNA structure by stabilizing the base pairing between the 5′ and 3′ UTR's. This “closed” structure of p53 mRNA might inhibit its translation by affecting the access of general translational machinery to the message. Upon stress stimulation, RPL26 binds to this double-stranded RNA structure containing the two UTR's and occupied by NCL, leading to a disruption of NCL dimerization and enhancement of p53 translation.

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 3086. doi:10.1158/1538-7445.AM2011-3086
Abstract 3095: The molecular, cellular, and physiological consequences of disrupting the HIF-1α interaction
Authors: Reece Kelie M. , Pisle Stephen T. , Richardson Emily D. , Cook Kristina M. , Price Douglas K. , Figg William D.
Published: June 2012

Many cancers contain regions of hypoxia due to rapid cell proliferation and the presence of blood vessels within the tumor that are structurally and functionally abnormal, resulting in the heterogeneity of blood flow. This state of hypoxia is associated with increased risk of treatment failure, metastasis and patient mortality. A principal mechanism by which cancer cells adapt to the hypoxic microenvironment is through the activity of hypoxia-inducible factor 1 (HIF-1), a heterodimeric transcription factor composed of an O2-regulated HIF-1α subunit that dimerizes with a constitutively expressed HIF-1β subunit. HIF-1α expression under hypoxic conditions regulates hundreds of genes including those that play key roles in invasion/metastasis, angiogenesis, tumor cell immortalization, and resistance to chemotherapy and radiation. Therefore, the inhibition of transcription driven by HIF has the potential for cancer treatment. One way to reduce the activity of this protein is by disrupting the complex that HIF forms with p300, an essential transcriptional coactivator. Previous data from our laboratory showed that several members of the epidithiodiketopiperazine (ETP) family of natural products are able to block the interaction between HIF-1α and p300. Here, we extend these studies by examining the comprehensive effect that select ETPs have on the disruption of the HIF/p300 complex at the molecular, cellular, and physiological levels. We began our studies by performing a series of rat aortic ring assays to determine the antiangiogenic effect of selected ETPs, namely chetomin, chaetocin, and gliotoxin. Chetomin and chaetocin concentrations of 50 nM inhibited approximately 90% of outgrowth, while 500 nM of gliotoxin was needed to achieve a similar effect; these compounds had GI50 of 22, 11, and 175 nM, respectively. In ongoing work, we hope to determine if the antiangiogenic effects that were observed in our rat aortic ring assays, are due to disruption of the HIF/p300 complex. To this end we are immunoprecipitating the complex from cells in the absence or presence of the aforementioned ETPs. The molecular consequences of blocking the HIF/p300 interaction will be assayed by qRT-PCR for hypoxia-regulated genes such as VEGF and GLUT1; cell proliferation and migration assays will also be performed to monitor cellular effects. Together, these results will be key in determining the feasibility of targeting this interaction for cancer therapeutics.

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 3095. doi:10.1158/1538-7445.AM2011-3095
Abstract 314: Distinctive genomic signature patterns of common Hematological malignancies uncovered by chromosomal microarray analysis (CMA) using a cancer-specific microarray
Authors: Li Marilyn M. , Hu Xiaofeng , Freeman Christopher E. , Carmack Condie E. , Iqbal Anwar
Published: June 2012

The application of chromosomal microarray analysis (CMA) in cancer research has produced a wealth of useful information about copy number alterations (CNAs) and their implications in cancer classification, disease progression, therapy response, and patient outcome. However, only a handful of clinical laboratories are offering CMA for cancer diagnosis. This is largely due to the lack of effective diagnostic standards and guidelines for cancer applications, and the absence of a cancer microarray database to aid in post-analytic interpretation. We have designed a combined targeted-/whole-genome array specific for cancer using the microarray CGH format. Approximately 20,000 high quality oligonucleotide probes were selected from the Agilent Tech. (Santa Clara, CA) eArray system to target all exons and exon/intron boundaries of more than 500 cancer genes, over 100 known cancer-associated genomic regions and all subtelomeric chromosome regions. Intervals between aforementioned genes or regions were filled relatively evenly with oligonucleotide probes to cover the whole genome. Using this custom designed array, we studied 250 cases of hematological malignancies and 30 normal bone marrow or blood controls. In addition to confirming and clarifying cytogenetic and FISH results, the array revealed many previously unknown CNAs, including intra-gene deletions and duplications in patients with normal or abnormal karyotype. Many apparently balanced translocations were found to harbor cryptic CNAs at or near the breakpoints. Many CNAs displayed strong association with specific category of malignancies. GeneSpring Hierarchical Clustering analysis showed distinctive genomic signature patterns for different types or sub-types of haematological cancers. Using BlueFuse Multi software, we were able to create a few disease classification/decision tracks, which are of great clinical significance in post-analytic interpretation and genotype-phenotype correlation for cancer.

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 314. doi:10.1158/1538-7445.AM2011-314
Abstract 3145: Comparing the reproducibility and prognostic value of proliferation markers Ki67, MAI, PPH3 in node negative breast cancer
Authors: Baak Jan P. , Gudlaugsson Einar , Skaland Ivar , Malpica Anais , Zhimin Shao , Janssen Emilius
Published: June 2012

Background. The proliferation factor Ki67 is prognostic but determination methods differ. Their reproducibility and independent prognostic value is largely unknown.

Aim. To evaluate the reproducibility of different Ki67 assessment methods and compare their prognostic value with Mitotic Activity Index (MAI), Phosphohistone-3 (PPH3) and other prognosticators.

Methods. In 240 T1,2N0M0 breast cancers without systemic adjuvant treatment, standardized automated quantitative immunohistochemistry was used for Ki7 and PPH3 and a strictly formalized protocol for MAI assessment. The percentages of Ki67 positive nuclei were assessed by two pathologists, computerized interactive morphometry (CIM) and fully automated digital image analysis (DIA). Reproducibility was studied by duplicate blind assessments. Continuous variables were discretized. We analyzed the prognostic value of widely used Ki67 thresholds (15%, 20%), others). We also tested at which places within the tumour (periphery, center), Ki67 was prognostically strongest. Univariate and multivariate survival analyses were performed.

Results. The Ki67 results of the two pathologists differed and their optimal prognostic thresholds greatly varied (4% and 14%). Moreover, Ki67% by one pathologist was well reproducible but not strongly prognostic, the other was strongly prognostic but not well reproducible. On the other hand, subjective Ki67% assessments were strongly prognostic (<0.0001) over a wide range of Ki67% thresholds (between 5% and 20%) (with an optimum Hazard Ratio for thresholds between 10%-14%). DIA-Ki67 and CIM-Ki67 were very well reproducible (r>0.99 and 0.94) and also strongly prognostic (P<0.0001 for both methods). Ki67 determinations are prognostically strongest in the periphery of the tumor. DIA-Ki67-6.5 is the strongest prognosticator, overshadowing subjective Ki67 assessments, Ki67-15% and 20% thresholds, tumor diameter, grade, ER, PR, Her2, CK5-6. CIM-Ki67 is a cheap well reproducible strongly prognostic alternative. MAI (<3 versus ≥3) or PPH3 (<13 versus ≥13) were slightly prognostically stronger than Ki67-DIA and considerably stronger than Ki-67-CIM.

Conclusions. In node negative breast cancer without adjuvant systemic treatment, Ki67 by digital image analysis or computerized morphometry but not by subjective counts is well reproducible and prognostically strong. MAI or PPH3 were prognostically slightly stronger.

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 3145. doi:10.1158/1538-7445.AM2011-3145
Abstract 3153: Detecting PTEN deletions in prostate cancer using circulating tumor cells
Authors: Atwal Siminder Kaur , Savage Heidi , Mohan Sankar , Pandita Ajay , Hampton Garrett , Amler Lukas , Lackner Mark , Punnoose Elizabeth
Published: June 2012

The PI3K/Akt pathway is frequently activated in human cancers. A number of the components of this pathway are activated, mutated, amplified and deleted in a wide variety of human cancers highlighting the key role of this pathway. PTEN is a negative regulator of this pathway that is frequently inactivated in a variety of tumors. PTEN loss could be useful as a predictive biomarker to identify the tumors that are dependent on the PI3K/Akt pathway and therefore likely to respond to PI3K/Akt targeted agents.

PTEN is frequently deleted in prostate cancer with incidence as high as 75% reported in advanced disease. Both genetic models of PTEN loss in prostate cancer as well as associations of PTEN deletions with poor clinical outcome support a role for PTEN in prostate cancer progression. Determining PTEN status is challenging in metastatic prostate cancer since biopsies are infrequently performed in this setting. Circulating tumor cells (CTCs) could provide an alternate non-invasive source of tumor cells and relatively higher counts of CTCs have been reported in prostate cancer compared to other indications. Here we evaluate the utility of CTCs in determining PTEN status of patients with castration resistant prostate cancer (CRPC).

We evaluate whether we can detect PTEN loss in CRPC patient CTCs and in their respective tissue. We developed a PTEN FISH assay to score PTEN in spiked CTCs in blood and compared PTEN loss in CTCs (captured on the Cellsearch platform) with archival tissue available from CRPC patients. Our data suggests that predictive biomarker analysis in CTCs is possible and CTCs could be potentially used to predict PTEN loss.

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 3153. doi:10.1158/1538-7445.AM2011-3153
Abstract 3155: Identification of biomarkers of chemoresistance in serous epithelial ovarian cancer by integrative molecular profiling
Authors: Koti Madhuri , Vidal Ricardo , Haslehurst Alexandria , Nuin Paulo , Weberpals Johanne , Childs Timothy , Bryson Peter , Feilotter Harriet , Squire Jeremy , Park Paul C.
Published: June 2012

Development of primary resistance to carboplatin and paclitaxel pose a major challenge in the management of ovarian cancer. To identify the molecular mechanisms underlying this process, we used microarrays to profile the 1) copy number alteration and SNP, 2) mRNA, 3) miRNA and 4) methylation signatures in 11 chemoresistant and 13 sensitive tumour samples, as defined by the RECIST criteria. The profiles are analyzed by Bayes statistics, based on R/Bioconductor packages, and the relevant pathways determined using the Ingenuity Pathway Analysis. The data from each array platforms are integrated using bioinformatic analytical and visual tools developed in house to not only decipher the most critical biological pathways, but also to identify the molecular mechanisms driving the pathways. Analysis to date identified the metabolic network involving HNRNPC, JAK1, Erbb2, ARF1, among others, which converge to deregulate the PI3K pathway. Interestingly, this is consistent with PTEN loss which is frequently observed in serous low grade tumours. The complexity of this pathway is reflected by its implication in multiple biological functions including growth promoting pathways, proliferation, differentiation, anti-apoptosis, tumorigenesis and angiogenesis. The integrated analysis will dissect and elucidate the roles that the CNA, SNP, methylation and miRNA play in the deregulation of this and other pathways involved in primary chemoresistance. Our research findings will yield diagnostic and prognostic biomarkers that will lead to development of specific treatment regimens for the improved control of serous epithelial ovarian cancer.

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 3155. doi:10.1158/1538-7445.AM2011-3155
Abstract 3160: Clinical utility of circulating tumor cell monitoring for therapeutic response in gastric cancer patients
Authors: Hashimoto Yuuri , Uno Futoshi , Tsujino Yukio , Urata Yasuo , Kagawa Shunsuke , Fujiwara Toshiyoshi
Published: June 2012

Background: Circulating tumor cells (CTCs) in the peripheral blood are considered as a useful biomarker for disease outcome and treatment response because the presence of CTCs is associated with short survival. We previously established the biological imaging system to detect viable CTCs among millions of peripheral blood leukocytes using telomerase-specific replication-selective adenovirus expressing green fluorescent protein (GFP) (Kojima et al, J Clin Invest, 2010). In present study, we demonstrate the clinical potential of our method for monitoring the efficacy of treatment in patients with gastric cancer.

Methods: We used a GFP-expressing adenovirus variant, in which the telomerase promoter regulates viral replication (OBP-401), to selectively label human tumor cell with fluorescence. Whole blood cells collected from 7.5ml of peripheral blood were infected OBP-401 after elimination of anti-adenovirus antibodies and incubated at 37 oC. Viruses were inactivated 24 hours after virus-infection and then red blood cells were lysed. The GFP-expressing cells were detected using fluorescence microscopy.

Results: A total of 72 blood samples from 39 patients with histologically-confirmed gastric cancer (stage IA to IV) were analyzed. CTCs were detected in 32 of 39 patients (82%, range 1 to 40<) before treatment, however, there was no apparent relationship between the number of CTCs and clinical status. We further assessed the CTC dynamics in 24 patients who were undergoing surgery, chemotherapy or endoscopic submucosal dissection (ESD). In patients with early gastric cancer, 7 patients had decrease in CTC numbers after surgery or ESD, and 6 patients had decreased CTC numbers following temporary increase after treatment. As for advanced cancer, the number of CTCs dropped in 3 patients after complete resection and increased in 2 patients with poor prognosis despite chemotherapy.

Conclusion: Our data suggest that patients with early gastric cancer also have CTCs in peripheral blood and the dynamic change of CTCs correlate with treatment response. Although numerous samples and long-term outcome should be analyzed, the enumeration of CTC using this GFP-expressing virus-based method might be useful for monitoring the efficacy of treatment.

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 3160. doi:10.1158/1538-7445.AM2011-3160
Abstract 318: Hierarchical clustering analysis of array CGH data identifies clinicopathologically distinct groups of lymph-node-negative invasive breast cancer
Authors: Kikuchi Ryoko , Ishii Yasuyuki , Kobayashi Takayuki , Kanai Yae , Shibata Tatsuhiro , Matsubara Osamu , Imoto Issei , Inazawa Johji , Tsuda Hitoshi
Published: June 2012

If high-risk group of lymph node-negative (pN0) invasive breast cancer is accurately identified by a molecular diagnosis of biopsy or surgically resected tumor specimens, it would be beneficial for choosing the most appropriate adjuvant or neoadjuvant therapies.

In this study, we examined genomic copy-number alterations of the 800 cancer-related genes in 51 pN0 primary invasive breast cancers (20 relapsed and 31 non-relapsed cases) surgically resected from 51 patients at the NCCH, Tokyo, between 1990 and 1994. DNA isolated from acetone-fixed paraffin-embedded tissues using microdissection were labeled directly with fluorescence dyes and subjected to array-based comparative genomic hybridizaton (array-CGH). Although copy number differences between relapsed and non-relapsed cases were unclear, unsupervised hierarchical clustering analysis was by using these 800 cancer-related genes resulted in identification of three cluster groups with significant differences in clinical outcome. Group 1 was mostly (9 of 13) composed ER(−)/HER2(−) cases, group 2 comprised 9 ER(+)/HER2(−), 8 ER(+)/HER2(+), and 6 ER(−)/HER2(+) cases, and group 3 was mostly (12 of 15) composed of ER(+)/HER2(−) cases. By the log-rank test for overall survival (OS), group 1 showed significantly worse outcome than group 2 (p = 0.0138), but no significant difference was observed in relapse-free survival (RFS) between these groups (p = 0.17). Among 20 patients who suffered relapsed later, both OS and RFS rates of group 1 were significantly higher than those of group 2 (p = 0.0083 and 0.0018). Representative genes that amplified in group 1 were CCND2(12p13.32), CDKN1B(12p13.1), whereas those kind genes in group 2 were ERBB2(17q12), NGFR(17q21.33).

This work demonstrates that hierarchical clustering of array-CGH data by using 800 cancer related genes can classify pN0 invasive breast cancers into clinically relevant groups and might pave the way to develop a new tool to estimate the risk of relapse and death in patients with pN0 breast cancer.

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 318. doi:10.1158/1538-7445.AM2011-318
Abstract 3181: The combined use of urine and serum biomarkers in early detection strategies for pancreatic cancer
Authors: Nolen Brian M. , Lomakin Aleksey , Grizzle William E. , Jhala Nirag , Eloubeidi Mohamad A. , Christein John D. , Arnoletti Juan P. , Oelschlager Denise K. , Vickers Selwyn M. , Lokshin Anna E.
Published: June 2012

The devastatingly high level of mortality associated with pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death in the United States, has remained virtually unchanged over the past 50 years due to an inability to effectively detect and treat the disease at an early stage. Currently used imaging techniques including CT and MRI have demonstrated an inability to distinguish benign from malignant pancreatic lesions resulting in both a low sensitivity for pancreatic cancer and a high degree of unnecessary referral for highly invasive endoscopic ultrasound (EUS). A biomarker-based test that could effectively rule out the possibility of malignancy in patients presenting with an abnormal finding by CT or MRI would greatly reduce the number of unnecessary EUS procedure and the associated risk of complication as well as lead to an optimization of resources for more effective early detection of the disease. Serum biomarkers such as CA 19-9 have been under investigation in pancreatic cancer for a number of years. Recently, the use of urine in biomarker studies has garnered considerable interest based on its ease of sampling, high stability, and relative simplicity as a testing matrix. In order to evaluate the use of urine and serum biomarkers in pancreatic cancer screening, we analyzed 77 cancer-associated biomarkers in urine obtained from 55 patients diagnosed with PDAC and 44 patients diagnosed with benign pancreatic conditions. Twenty of the most relevant biomarkers were also tested in sera obtained from the same patient cohort. Forty of the 77 urine biomarkers as well as nine of the twenty serum biomarkers were significantly altered between the benign and malignant groups. Of the twenty biomarkers measured in both urine and serum, seven were more significantly altered in urine, while seven others were more significantly altered in serum. A multivariate analysis of the results identified a four-biomarker panel comprised of both serum and urine biomarkers which could discriminate PDAC from benign disease with optimal levels of sensitivity and specificity. Our results indicate a promising role for the combined use of urine and serum biomarkers in the development of improved screening strategies for pancreatic cancer.

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 3181. doi:10.1158/1538-7445.AM2011-3181
Abstract 3182: PTM ELISA microarray for breast cancer biomarker discovery
Authors: Jin Hongjun , Daly Don S. , Tan Ruimin , White Amanda M. , Marks Jeffrey R. , Zangar Richard C.
Published: June 2012

Post-translational modifications (PTMs) of proteins are known to be altered during breast cancer development. These PTMs are potentially useful biomarkers for breast cancer. In order to study the potential of PTMs in the early detection of breast cancer, ELISA microarray protocols for analyzing several PTMs in a panel of 24 proteins that are found in blood but known to be secreted by breast tissue and/or breast tumors. We evaluated these PTMs in a case-control study with 68 plasma samples. The control samples were from age-matched women with benign breast disease. The sensitivity and the specificity of each assay and the combined assays were evaluated using the areas under (AUC) the receiver operator curves. In the best multivariate analysis, the AUC values ranged from 77% to 87% for 4 different PTMs. Overall, our data suggest that the PTM ELISA microarray platform is a promising tool for discovery and evaluation of biomarkers that have potential for the early detection of breast cancer.

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 3182. doi:10.1158/1538-7445.AM2011-3182
Abstract 320: Association of higher Gleason grade with presence of PTEN deletion in prostatic adenocarcinoma
Authors: Yoshimoto Maisa , Evans Andrew , Sweet Joan , Ludkovski Olga , Ding Keyue , Trottier Greg , Song Kyu S. , Squire Jeremy A.
Published: June 2012

Prostatic adenocarcinoma (CaP) is one of the most histologically heterogeneous of epithelial malignancies. The Gleason score is widely accepted as the standard for histologic grading and determining prognosis of prostate malignancies. The tumor often presents with a multifocal distribution though the gland and different Gleason grades may be present in different foci. The current study was undertaken to compare the association of genomic alterations with the Gleason grades of individual adenocarcinoma foci. Genomic deletion of the PTEN gene in routine formalin-fixed paraffin-embedded (FFPE) pathology sections can be readily assayed by fluorescence in situ hybridization (FISH). Data from our group and others has shown that the genomic deletion of the PTEN tumor suppressor gene and the disruption of the ETS gene family, play a central role in prostate cancer and are likely to be associated with Gleason grade. In this study CaP samples were thoroughly analyzed to identify PTEN heterogeneity, within or between foci in multifocal tumors. This study was designed to retrospectively assess the overall frequency with which PTEN deletions and TMPRSS2-ERG fusion rearrangements occur individually and simultaneously, and their clinical impact through FISH analysis using well-annotated high-throughput tissue microarrays (TMAs) derived from 128 patients with CaP. In addition, four-color FISH analysis of the PTEN deletion was used to precisely detect the interstitial deletion of genomic content from this region of chromosome 10q and evaluate the tumor heterogeneity. Genomic PTEN deletion was observed in 55 of 128 (43%) patients with CaP (hemizygous = 41 and homozygous = 14 CaP). TMPRSS2-ERG fusion was observed in 58 of 129 (45%). We specifically examined the relationship between PTEN deletion and TMPRSS2-ERG fusion status, Gleason pattern and clinical behavior of CaP. Importantly, PTEN deletions were significantly associated with a higher primary Gleason pattern grade 4 (p=0.000), lymphovascular invasion (p=0.051), and higher percentage of prostate involved by tumor (p=0.0064). Although TMPRSS2-ERG fusions showed a strong relationship with PTEN deletions (p=0.007), TMPRSS2-ERG fusions did not show any clinical correlation. In conclusion, our findings indicated that PTEN deletion FISH can be applied to CaP samples to accurately stratify patients into risk categories and to guide decision-making with respect to the optimal management of CaP 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 320. doi:10.1158/1538-7445.AM2011-320
Abstract 3219: Isolated tumor cells (ITC) in regional lymph nodes predict colorectal cancer (CRC) relapse
Authors: Mescoli Claudia , Albertoni Laura , Fassan Matteo , Lazzarin Vanni , Lanza Cristiano , Giacomelli Luciano , Pucciarelli Salvatore , Nitti Donato , Rugge Massimo
Published: June 2012

Involvement of the lymph nodes (LN) is the most important prognostic factor in CRC and pN+ve status identifies patients requiring adjuvant chemotherapy. Among pN0-patients, 20-30% develop recurrent disease. ITC+ve LNs in such patients (as assessed by immunohistochemistry) may potentially predict cancer recurrence. No prospective long-term follow-up trial has consistently validated the prognostic meaning of ITCs.

ITCs in regional LNs were prospectively assessed in a cohort of 361 consecutive pN0-CRC patients. From each gross surgical specimen, LNs were microdissected according to a standardized protocol (mean LNs per patient=16.4; range 5-107). In all, 5,920 pN0-ve LNs were collected and immunohistochemically assessed (MNF116 staining was performed in 2 serial histology sections of each LN [11,840 histology specimens in all]). Overall cancer death and recurrence rates were calculated. The prognostic impact of ITC was also tested by multivariate statistics.

ITCs were documented in 200 of 361 patients (55.5%). No relationship was documented between ITC+ve status and overall survival. CRC recurred in 36/361 (9%) cases and 29/36 (80%) recurrences were documented among ITC+ve patients. The prevalence of recurrent CRC among ITC+ve and ITC-ve patients was 14.5% and 4.3%, respectively. ITC+ status significantly correlated with cancer recurrence (p = 0.002, OR = 3.57; 95% CI=1.46-9.95; case-control Pearson's chi-squared) and disease-free interval (p=0.0005). In multivariate analysis, ITC-status was the only variable significantly associated with cancer recurrence.

These results suggest considering ITC+ve status a reliable marker of cancer recurrence, and that pN0 ITC+ve cancer patients form a subgroup of candidates for adjuvant chemotherapy.

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 3219. doi:10.1158/1538-7445.AM2011-3219
Abstract 322: Triple combination array analysis detects the Collagen type1 alpha1 (COL1A1) gene as a new tumor related gene in hepatocellular carcinoma
Authors: Masamichi Hayashi , Nomoto Shuji , Okamura Yukiyasu , Hishida Mitsuhiro , Nishikawa Yoko , Fujii Tsutomu , Sugimoto Hiroyuki , Takeda Shin , Nakao Akimasa
Published: June 2012

Background: Epigenetics is one of the most important mechanisms for inactivating tumor suppressor genes or tumor-related genes in various human cancers. Hepatocellular carcinoma (HCC) is one type of human cancer in which epigenetic gene suppression is frequently found. To effectively detect a tumor-related, we performed triple combination array analysis, which consists of an expression array, a single nucleotide polymorphism (SNP) array, and a methylation array of the same surgical HCC specimen.

Methods: Triple array analysis of one HCC sample from a 68-year-old female patient was performed. Gene expression profiles were determined using Affymetrix HGU133A and B Gene-Chips using RNA. The SNP chip array was conducted using the standard protocol for Affymetrix Gene Chip Mapping 500K arrays. Methylation array analysis used an Illumina Infinium HumanMethylation27 Beadchip Kit. The gene encoding Collagen type1 alpha1 (COL1A1) was identified and verified using HCC cell lines and 48 primary HCC tissues from patients who had undergone hepatectomy at our hospital.

Results: Expression array analysis revealed that COL1A1 gene expression was decreased in tumor tissues (Log2 Ratio -1.1). In addition, SNP array analysis showed that the COL1A1 locus, 17q21, did not show a chromosomal deletion. Importantly, the methylation value (0-1.0) in the tumor tissue was higher (0.557) than that of the adjacent normal tissue (0.008). We hypothesized that the expression of this gene was suppressed by promoter hypermethylation. The suppressed expression of the COL1A1 gene in HCC cell lines was reactivated after 5-aza-2′-deoxycytidine (5-aza-dC) treatment. Sequence analysis identified hypermethylated CpG sites in the COL1A1 promoter region. In 48 HCC samples, 21 cases (44%) also showed COL1A1 gene promoter hypermethylation. The expression values of methylated cases, examined by real-time RT-PCR, were significantly downregulated (p=0.004) compared with unmethylated cases. Downregulation of the gene's expression was significantly correlated with pathological fc positive (p=0.026) and liver damage scores (p=0.011).

Conclusion: Triple combination array analysis successfully revealed that the COL1A1 gene acts as a tumor suppressor gene in HCC via promoter hypermethylation.

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 322. doi:10.1158/1538-7445.AM2011-322
Abstract 3228: Preparation and in vivo evaluation of a phytosphingosine derivative, N,N,N-Trimethylphytosphingosine (TMP), in liposomes for the treatment of tumor metastasis
Authors: Song Chung Kil , Balakrishnan Prabagar , Shim Chang-Koo , Chung Suk-Jae , Chong Saeho , Choi Myeong Jun , Kim Dae-Duk
Published: June 2012

Purpose: The aim of this study is to evaluate the cytotoxicity and anti-metastatic effect of a phytosphingosine derivative, N,N,N-trimethylphytosphingosine.iodide (TMP-I), and to compare it with that of TMP-I liposome. Background: Sphingolipids are known for ortho-signaling and anti-adhesion features as well as intracellular signal messenger in cell differentiation, proliferation and apoptosis. Sphingosine and sphinganine, structural analogs of sphingolipids were recognized as negative modulator of transmembrane signaling through protein kinase C (PKC) that plays an important role in the cancer treatment. Among them, phytosphingosine has been known for antitumor and anti-invasive effect through phytosphingosine-induced apoptosis mechanism. Recently it has been reported that N-monomethylphytosphingosine and N,N-dimethylphytosphingosine induced apoptosis via mitochondria-involved apoptosis such as caspase-8, caspase-3 and caspase-9 and this prompt us for this study. Methods: TMP-I solution was evaluated for cytotoxicity in various cancer cell lines and hemolysis in rat blood cells at various concentrations. The prepared TMP-I liposomes (DPPC:Chol:TMP-I at 5:5:1 mole ratio) were characterized by particle size and studied for in vitro cytotoxicity, cell migration and in vivo angiogenesis in rats. Furthermore, to evaluate the anti-metastatic effect of TMP-I liposomes, C57BL6 mice (n=5) were inoculated with B16F10 cells in tail vein for direct metastasis and footpad for spontaneous metastasis studies. Results: The prepared TMP-I liposomes were with a mean size of 150 nm. TMP-I solution showed significantly higher cytotoxicity and hemolysis (≥100 μg/ml) compared to that of TMP-I liposomes in several cancer cell lines. TMP-I liposomes showed significantly low cell migration and angiogenesis compared to control formulations. The in vivo study in mice showed that the numbers of lung nodules in the mice treated with TMP-I liposomes were significantly lower compared to that of control formulations. Conclusion: Our results suggested that the TMP-I liposome could be used as a potential carrier for the treatment of tumor metastasis.

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 3228. doi:10.1158/1538-7445.AM2011-3228
Abstract 3238: Synthesis and evaluation of 17-oximino-estrone derivatives as potential anti-breast cancer agents
Authors: Joseph Patrick , Cooperwood John S. , Wan Zhongliang
Published: June 2012

The selective estrogen receptor modulator (SERM) tamoxifen (TAM) is the gold standard used in the treatment of estrogen-dependent breast cancer. Although it significantly benefited women with breast cancer, time-dependent tumor resistance and the risk of endometrium cancer are associated with it. Consequently, there is an urgent need for alternative treatments. PURPOSE: Incorporating the oxime moiety in conjunction with various basic side chains associated with the pharmacophore of the SERMs into estrone may yield various estrone derivatives with antiestrogenic activity. The estrone derivatives were compared to 4-hydroxy-tamoxifen (4-OH-TAM) and raloxifene (RAL). METHODS: The estrone derivatives were synthesized using established chemical procedures. The Southern Research Institute in Birmingham, Alabama carried out the cytotoxicity studies in MCF-7, MDA-MB-231 and Ishikawa cancer cells. They tested different concentrations of the six different estrone derivatives ranging from .01 nM to 100,000 nM on 5000 MCF-7 cells/well, on 5000 MDA-MB-231 cells/well and on 5000 Ishikawa cells/well. Cells pre-treated with the various estrone derivatives were incubated at 37°C for 3 days followed by MTS assay. Their IC50 values were generated using GraphPad Prism 5.0. RESULTS: Our compounds demonstrated anti-proliferative activity in MCF-7 cells but they were not as potent as 4-OH-TAM(2µM) or RAL (3µM). Compounds 2, 3, 4, 6, and 7 (31µM, 29µM, 30µM, 30µM, and 29µM respectively) were comparable in potency to RAL (28µM) in MDA-MB-231 cells but all were not as potent as 4-OH-TAM (22µM). Compounds 3, 6, and 7 (20µM, 20µM, and 17µM respectively) were comparable or greater in potency to 4-OH-TAM (20µM) in Ishikawa cells and compounds 2, 3, 6, and 7 (24µM, 20µM, 20µM, and 17µM respectively) were comparable or greater in potency to RAL (23µM). CONCLUSION: Applying the oxime moiety in conjunction with the pharmacophore of the SERMs to estrone produced agents with antiproliferative activity. They also showed antiproliferative activity in estrogen-receptor negative MDA-MB-231 cells. These results suggest that several of the estrone derivatives show promise as potential anti-breast cancer agents.

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 3238. doi:10.1158/1538-7445.AM2011-3238
Abstract 3243: Discovery of novel Rho kinase inhibitors: Hit generation and lead optimization
Authors: Alexander Brian G. , Pireddu Roberta , Sun Nan N. , Yu Huidong , Forinash Kara D. , Martin Matthew P. , Schonbrunn Ernst , Guida Wayne C. , Sebti Said M. , Lawrence Nicholas J.
Published: June 2012

Rho associated protein kinases (ROCKs) are Ser/Thr kinases which require activation by small GTPases of the Rho family. ROCK is often found to be over expressed in ovarian, testicular, bladder, and pancreatic cancers. ROCK has been implicated in cell motility, adhesion, smooth muscle contraction and stress fiber formation. Inhibition of Rho kinase is a novel approach for the treatment of cardiovascular disease, CNS disorders, and, more recently, cancer. ROCK has received increased attention for its affect on cell adhesion and invasion, two key steps in metastasis. We detail the development of ROCK inhibitors from initial small molecule library screening and in-house HTS screening data. We report on the identification and resynthesis of one screening hit (ROCK1 IC50 38.5 ± 3.4 µM). Focused chemical libraries were developed and structure activity relationships will be described. All compounds have been screened in a Z-lyte kinase assay to determine their activity against both ROCK1 and ROCK2. Hit-to-lead optimization and SAR studies resulted in compounds capable of inhibiting ROCK1 at low nanomolar concentrations, RPM1510 (ROCK1 IC50 0.15 ± 0.02 µM) and RPM1533 (ROCK1 IC50 0.12 ± 0.02 µM). Progress in obtaining structural information by X-ray crystallography will be reported along with ROCK inhibitory data. Trends in isoform and kinase selectivity and potency will also be discussed. Members of this class have been shown to inhibit potently, in human lung cancer cells, the phosphorylation of MYPT1, a surrogate for ROCK activity in vivo.

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 3243. doi:10.1158/1538-7445.AM2011-3243
Abstract 3267: Inhibition of angiogenic potential of neuroblastoma cells through regulation of microRNA expression profiles by tumor suppressor gene dipeptidyl peptidase IV
Authors: Lee Raymond , Dempsey Robert J. , Wesley Umadevi V.
Published: June 2012

Neuroblastoma (NB), a childhood cancer of peripheral nervous system generally progress to highly angiogenic and therapy resistant fatal metastatic tumor. We have recently shown that a cell surface protease called dipeptidyl peptidase (DPPIV) is expressed in normal neural crest derived differentiated cells and its expression is lost in proliferating NB, the malignant counterpart. We also demonstrated that re-expression of DPPIV in NB cells leads to their differentiation, apoptosis, and suppression of their migratory and angiogenic potential. Importantly, DPPIV inhibits the tumorigenic potential of NB cells in a xenograft mouse model. To further identify DPPIV-responsive genes and mechanisms, we used Affymetrix microarray analysis. This study showed an association of DPPIV expression with up-regulation of thrombospondin-1 (THBS1), an angiogenesis inhibitor. Interestingly, some of the genes regulated by DPPIV are also altered by microRNA (miRNAs), indicating that at least some of the miRNAs might act as intermediate effectors in mediating DPPIV function. PCR array screening of cancer specific miRNAs, showed that DPPIV indeed alters miRNA profile in NB cells. In particular, DPPIV down-regulated miR-19a which is a part of the miR-17-92 cluster that is known to decrease THBS1 levels. Furthermore, bioinformatics data mining of Sangers MicroCosm Targets v5 predicted THBS1 as a potential target of miR-19a suggesting that down-regulation of miR19a leading to up-regulation of THBS1 was a possible mechanism involved in DPPIV tumor suppressor function. In support of this idea, silencing of DPPIV expression by RNA interference resulted in increased miR-19a and decreased THBS1 expression. Furthermore, transfection of NB cells with anti-miR-19a decreased miR19a and increased THBS1 levels as shown by real time quantitative RT-PCR, immunofluorescence and western blot analysis. Furthermore, DPPIV expression decreased the levels of cMYC, a transcription factor that up-regulates miR-19a expression. These effects were associated with decreased levels of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), two strong downstream pro-angiogenic factors. Taken together, our work identifies a novel and an important mechanism wherein DPPIV inhibits angiogenesis and growth of NB tumor cells through down regulation of miR-19a mediated c-MYC expression and increased expression of THBS1, an angiogenesis inhibitor.

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 3267. doi:10.1158/1538-7445.AM2011-3267