bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3059: CCL2 overexpression in an epithelial ovarian cancer cell line results in reduced tumorigenic potential
Authors: Wojnarowicz Paulina M. , Gambaro Karen , Desaulniers Kim Leclerc , de Ladurantaye Manon , Provencher Diane , Mes-Masson Anne-Marie , Tonin Patricia N. 
Published: June 2012

Abstract
Chromosome 17 abnormalities are frequently observed in epithelial ovarian cancer (EOC), suggesting that one or more genes mapping to this chromosome contribute to EOC development and/or progression. We have previously performed a comparative transcriptome analysis of primary cultures of normal ovarian surface epithelial (NOSE) cells and malignant, serous subtype ovarian tumor (TOV) samples in order to identify differentially expressed chromosome 17 genes. CCL2, at 17q12, was significantly differentially underexpressed at least 3-fold in all TOV samples. We have confirmed underexpression in an independent series of 99 serous EOC tumor samples. Interestingly, CCL2 was identified as a significantly up-regulated gene occurring as a consequence of tumor suppression in the characterization of a novel EOC cell line model developed by transferring chromosome 3 fragments into the tumorigenic OV-90 EOC cell line. We have further characterized CCL2 expression in a series of EOC cell lines that differ in their tumorigenic potential, and have shown the lowest levels of gene expression occur in tumorigenic cell lines. DNA sequencing did not reveal any evidence of mutations. To determine if overexpression affects tumorigenicity or growth characteristics, we have generated stable overexpressing OV-90 clones using a commercially available pDream: CCL2 expression vector. Overexpression was verified by ELISA. In cell culture, the CCL2 overexpressing clones appeared less tightly packed and more elongated as compared with OV-90 cells. The cell viability of the CCL2 clones appears to be reduced when compared to the parental OV-90 cell line in XTT cell proliferation assays. The ability of the CCL2 clones to form colonies was also reduced as compared to OV-90 in a colony formation assay. There is also evidence of an increase in the percentage of senescent cells in the CCL2 clones based on a beta-galactosidase staining assay. Both CCL2 overexpressing clones exhibit latency in tumor formation in intraperitoneal tumorigenicity assays involving SCID mice as compared with the parental OV-90 cell line. This is the first report of stable CCL2 transfection into an EOC cell line. Our results suggest that CCL2 expression may reduce the tumorigenic potential of EOC cells, and draws attention to the role of the innate immune system and the tumor microenvironment in EOC development and progression.
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 3059. doi:10.1158/1538-7445.AM2011-3059
Abstract 306: Development of EC-DG as a molecular theranostic personalized medicine
Authors: Yang David J. , Ford Richard J. , Mendez Richard , Zhang Yin-Han , Bryant Jerry , Oh Chang-Sok , Huang Jack , Pham Lan , Kohanim Saady , Kim Edmund E. 
Published: June 2012

Abstract
Purpose: D-glucosamine has been reported to inhibit proliferation of cancer cells in culture and in vivo. We have then synthesized Tc-99m- ethylenedicysteine-glucosamine (EC-DG). We found Tc-99m-EC-DG was involved in cell proliferation in lung, breast and head and neck cultures and could assess breast cancer treatment outcome in vivo by planar scintigraphy. Tc-99m-EC-DG is a safe imaging agent in lung cancer patients. This study was amied to (1) assess a novel response to unlabeled rhenium-EC-DG (Re-EC-DG) involving the translation regulation of hypoxia inducible factor (HIF)-1alpha expression in lymphoma cells, and, (2) evaluate feasibility of using EC-DG for theranostic approaches in cancers.
Methods: For theranostic assessment studies, we synthesized cold Re-EC-DG. Re-EC-DG was synthesized via a two-step synthesis. The first step was to synthesize Re-EC by reacting rheniumoxo trichloride with EC. The second step was to react Re-EC with D-glucosamine tetraacetate, followed by de-acetylation. Twelve types of DLBCL cells were incubated with Re-EC-DG at various concentrations (0-10 mM) and TUNEL assays were used to determine cell apoptosis. To ascertain the mechanism of the anticancer properties for Re-EC-DG, DLBCL-LY10 cells were treated with Re-EC-DG (0-5 mM) for 48 hrs. Immunoblotting were then performed on nuclear extracts with 50 µg. For radiotheranostic assessment studies, 13762 breast tumor-bearing rats were imaged with In-111-EC-DG and tumor/muscle ratios were determined at 0.5-24 hrs. Radiation absorbed dose was estimated for the use of Y-90-EC-DG.
Results: There was a dose response relationship of Re-EC-DG inhibition in DLBCL cells. Extensive apoptosis was observed at 24 hrs in lymphoma cell cultures. Re-EC-DG showed significant tumorcidal activity compared to normal B-lymphocyte activity at doses &gt;0.17 µmol. Re-EC-DG caused a decreased expression of HIF-1alpha under normoxic conditions in DLBCL-LY10 cells. Tumor-to-muscle ratios for In-111-EC-DG were 5.43±0.45 to 7.80±0.05 whereas In-111-EC had 3.24±0.32 to 4.64±0.16 at 0.5-24 hrs. Radiation exposure of In-111-EC-DG to whole body, blood-forming organs, gonads, and effective dose equivalent for a single dose at 5 mCi was below the limits of 3 rad annually and 5 rad total. The absorbed dose in all other organs was below the limits of 5 rad annually and 15 rad total.
Conclusion: EC-DG is a useful molecular theranostic compound. In-111-EC-DG has favorable dosimetry, providing a potential use of Y-90-EC-DG to treat cancers. Re-EC-DG inhibits HIF-1alpha expression and is an attractive anti-proliferation compound.
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 306. doi:10.1158/1538-7445.AM2011-306
Abstract 3060: Target genes of chromosomal 9p13.3 amplification in prostate cancer
Authors: Latonen Leena , Leinonen Katri A. , Saramäki Outi R. , Visakorpi Tapio 
Published: June 2012

Abstract
The development and progression of cancer is caused by alterations that accumulate in the genome of malignant cells. We are studying genetic alterations occurring in prostate cancer, the most common male malignancy in Western countries. We have previously identified a frequent (39%) gain at 9p13-q21 in 13 prostate cancer xenografts and 5 cell lines by aCGH (Saramäki et al., Int J Cancer 119:1322-1329, 2006). Preliminary results indicate that in clinical prostatectomy samples, gains and amplifications are found in approximately 30% and 10% of the cases, respectively, and that the amplification frequency seems to increase with disease progression to hormone-refractory stage. The 9p13.3 amplification is also found from several breast cancer cell lines, raising the possibility that the amplification target gene(s) in this area may take part in breast cancer formation in addition to prostate cancer.
We aim at identifying the target gene of the 9p13.3 amplification in prostate cancer. A minimal region for the 9p13.3 amplification, determined by fluorescence in situ hybridisation (FISH), spans nearly 3.5 Mb and contains more than 40 known and hypothetical protein-coding genes. We initially screened mRNA expression levels by quantitative RT-PCR and chromosomal copy number alterations by FISH for these genes in 7 prostate cancer cell lines and 19 prostate cancer xenografts. Thus, we narrowed down the list of amplification target candidates to eight protein-coding genes (C9orf25, GALT, PIGO, UBAP1, UBAP2, UBE2R2, UNC13B and VCP). We have performed siRNA-mediated downregulation of these genes in cell lines with either a normal copy number status or a gain/amplification in 9p13.3 (prostate cancer cell lines PC-3 and 22Rv1; breast cancer cell lines MCF-7 and BT-474). Downregulation of several of these genes decrease the proliferation rate of the cells significantly (GALT, PIGO, UBAP1, UBAP2, VCP). As decreased levels of VCP induce cell death in PC-3 cells, downregulation of UBAP1 induces a cell cycle arrest in the G2/M phase. In addition, invasion of PC-3 cells in Boyden chamber assay is affected by knock-down of GALT and PIGO. Genes exhibiting alterations in the siRNA assays have been further analyzed for their expression in clinical prostate cancer tumor material by qRT-PCR and immunohistochemistry. These genes are also studied by overexpressing their cDNAs in cell lines with normal 9p13.3 copy number status and screening for altered cellular growth, survival and invasion in vitro. We aim at identifying a novel diagnostic and/or prognostic marker for prostate cancer, and potentially also a target for new cancer therapies, from the 9p13.3 region.
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 3060. doi:10.1158/1538-7445.AM2011-3060
Abstract 3064: A novel method for mutation analyses using genomic DNA obtained from IHC-stained sections
Authors: Desai Rupal M. , Patel Rajesh , Amler Lukas , Stern Howard , Raja Rajiv 
Published: June 2012

Abstract
Biopsies obtained from patients are often limited in size. The ability to perform multiple diagnostic assays is dependent on the quantity and quality of tissue samples available from patients. Over the last few years, our laboratory has focused our efforts on developing novel methodologies for increased efficiency in use of tissue samples. Here we describe a method for performing immunohistochemistry (IHC) and genotyping using the same tissue section. We isolated genomic DNA (gDNA) from IHC-stained and unstained formalin-fixed paraffin-embedded (FFPE) tissue sections obtained from the same paraffin block. Quality and quantitylity of gDNA obtained from either sections were compared. We demonstrate that quality of gDNA obtained is acceptable for down-stream analysis using PCR techniques. We also show comparable specificity and sensitivity in detecting mutations using either sources of gDNA. This method has proven highly valuable in performing mutation analysis in certain tissue types such as lung cancer where tissue can be extremely limited.
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 3064. doi:10.1158/1538-7445.AM2011-3064
Abstract 3066: Direct detection and enumeration of circulating tumor cells (CTCs) in blood by CastRT-qPCR
Authors: Deng David X. , Sprout Scott , Bao Yun , Merrill David , Brzoska Pius , Wang Yu , Chen Caifu 
Published: June 2012

Abstract
Statistical data suggest that only approximately 25% of cancer patients will respond to the same treatment. Enumeration and molecular characterization of circulating tumor cells (CTCs) promise to be valuable for cancer cell diagnosis, survival prognosis, and treatment guidance. However, CTCs are very rare. There is no sensitive and specific assay available for analyzing CTCs from whole blood without extensive enrichment processes. Sample enrichment causes target cells loss, expensive and time-consuming. Here we report a highly specific competitive allele-specific TaqMan-based RT-qPCR (castRT-qPCR) for rare copies of DNA mutation detection. CastPCR can detect rare copies of mutant alleles for up to one in 10 million with a wide dynamic range of more than 6-log orders and &lt;5 copy sensitivity. We have successfully designed and validated cast-PCR assays for more than 80 cancer-related SNPs of KRAS, EGFR, BRAF, KIT, pTEN, p53 and other oncogenes or tumor suppressor genes. CastRT-PCR was used to specifically detect and enumerate CTCs in blood samples with lung cancer cell lines spiking-in and lung cancer patient blood without prior biochemical enrichment processes. EGFR mutation (p.L858R) was detected in all 5 blood samples from late stage lung cancers. Approximately 50% of circulating lung tumor cells with positive EGFR p.L858R mutation had positive EGFR p.T790M mutation. In normal blood samples spiked in cell lines of known Kras and EGFR mutation at the level of 10 cells/mL of whole blood (close to actual patient samples), CastRT-PCR accurately detected the known mutations. Our data suggest that digital castRT-qPCR can be used to enumerate CTCs and detect cancer-related mutations in the blood, which paves the way for future individualized cancer treatment by monitoring CTC level and molecular characterization.
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 3066. doi:10.1158/1538-7445.AM2011-3066
Abstract 3070: Simultaneous amplification of multiple genetic/epigenetic tumor markers in a single reaction for high throughput genotyping and aberrant methylation profiling
Authors: Liu Liansen , Liu Chunmei , Stevens Junko , Wang Jonathan T. 
Published: June 2012

Abstract
Matching appropriate therapeutic regimes with tumor genotypes and epigenotypes is of great medical and economical importance. A prerequisite for this approach is thorough and accurate genotype and epigenotype profiling of tumor specimens, which should be performed in precisely defined homogeneous tumor cell populations with minimum non-tumor cell contaminations. However, this can be very challenging due to the genetic and epigenetic heterogeneities and the scarcity of homogeneous tumor cells within a tumor specimen. Multiplex PCR allows high throughput profiling by maximizing the number of targets/reaction in each sample and can reduce the limitations of such sample shortage. Although several multiplex PCR kits have been commercially available for this purpose for some time, their usefulness is restricted by small amplicon size range, less satisfactory amplification uniformity of different amplicons, and occasional failure in amplifying some amplicons, or false positive amplifications. To solve these problems, we have devised a highly specific, optimization-free multiplex PCR formulation (Applied Biosystems Platinum® Multiplex PCR Master Mix) with novel reaction chemistry. We show here that this new formulation is excellent for simultaneous amplification and high throughput profiling of SNPs, deletions, insertions and their wild types of many targets in a single reaction in terms of robustness, uniformity, amplicon size range, compatibility with amplicon GC contents, and the reduction of non-specific primer dimers and false positive/negative products in multiplex PCR. In combination with a primer system of varied lengths and compositions of 5’ stem-loop tag sequences, we demonstrate that even mixtures of multiple adjacent or overlapping alleles in the same locus of EGFR/KRAS/BRAF mutations can be specifically amplified in the same reaction and distinguished by amplicon size differentiation in a single lane of an agarose gel. Moreover, using primer sets specific for methylated and converted DNA, and those specific for unmethylated and converted DNA, we demonstrate that EGFR/KRAS/BRAF mutations and their associated promoter methylation status can be amplified from bisulfite-treated genomic DNA of colon cancer cell lines in a single reaction. In conclusion, the Applied Biosystems Platinum® Multiplex PCR Master Mix and the stem-loop primer system are a “best in class” solution to simultaneous amplification of various types of genetic and epigenetic tumor markers of multiple targets for high throughput genotyping and aberrant methylation profiling.
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 3070. doi:10.1158/1538-7445.AM2011-3070
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

Abstract
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

Abstract
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