bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2728: Clonal nature of benign and malignant salivary gland neoplasms
Authors: Gomes Carolina Cavalieri , Pereira Claudia Maria , Brito Joao Artur R , Amaral Fabricio Rezende , Barbosa Alvimar Afonso , de Marco Luiz , Gomez Ricardo Santiago
Published: June 2012

The pathogenesis of salivary gland neoplasms (SGN) has not been established. There is a real debate on clonality issues and all the definitive evidence is on the side of polyclonal tumor origin. Evidence that many tumor types may be polyclonal in origin has been published. Despite all this body of evidence, scientists have incorporated the clonal theory proposed by Knudson (1985) and only strengthen these results (and do not discuss contrary findings), because they think it is expected. There are few clonality studies on benign salivary neoplasms and almost no data on malignant neoplasms. The HUMARA assay was used in the present study to investigate the clonality of 27 samples of SGN (17 benign and ten malignant). Briefly, samples were digested with HhaI and HpaII methylation-sensitive enzymes and a PCR carried out. Eighteen out of 27 cases revealed monotypy, while three were non-informative and six showed heterotypy, indicating a true polyclonality. In summary, our results show evidence that a subset of SGN might be clonal in origin, while some samples are polyclonal.

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 2728. doi:10.1158/1538-7445.AM2011-2728
Abstract 2745: Genetic determinants of telomere length – a genome wide association study
Authors: Chen Meng , Gu Jian , Ye Yuanqing , Lin Jie , Chang David , Wu Xifeng
Published: June 2012

Telomere plays a critical role in maintaining genome integrity. Telomere shortening is associated with the risk of many age-related diseases. Classical twin studies have shown that genetic components may contribute up to 80% of the heritability of telomere length. Previous association studies have reported several genetic loci in regulating telomere length, but the results have been inconsistent. In this study, we adopted a three-stage genome-wide association study (GWAS) design to identify the genetic determinants of telomere length. The mean telomere length was measured by quantitative real-time PCR. Genotyping were done by illumin's whole-genome SNP array in stage I and II, and by TaqMan assay in stage III. Multivariate linear regression analysis was applied to assess the association between genetic variants and telomere length adjusting for age and gender. In stage I, we analyzed 300K SNPs in 459 healthy controls and found 15,351 SNPs that were associated with telomere length at P<0.05. In phase II, we analyzed these 15,351 SNPs in two independent populations with GWAS data with 952 control subjects and 272 control subjects, respectively, and identified 18 SNPs, each with a combined P value < 10−4. Among them, two loci located at chromosome 20q11.22 and 14q21 were associated with telomere length with a significant P value < 10−7. We are in the process of genotyping these 18 SNPs and measuring telomere lengths in approximately 1,200 independent control subjects for stage III validation.

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 2745. doi:10.1158/1538-7445.AM2011-2745
Abstract 275: Burkitt lymphoma cells express GLI1 but are not responsive to Hedgehog signaling
Authors: Yoon Joon Won , Gallant Marisa , Lamm Marilynn , Leong King-Fu , Iannaccone Philip , Walterhouse David O.
Published: June 2012

Activation of the Hedgehog (HH) signaling pathway has been associated with a wide variety of human cancers. Some B-cell lymphomas express components of the HH signaling pathway, including patched (PTCH), smoothened (SMO), and GLI1. There is also evidence that HH ligands function as survival factors in these malignancies. However, specific roles for HH signaling in Burkitt lymphoma have not been characterized. Previously we showed that constitutive expression of c-MYC in Burkitt lymphoma cells up-regulates GLI1 and that GLI1 functions as an anti-apoptotic factor in this setting. Similar to other B-cell malignancies, we have shown expression of PTCH, SMO, and GLI1 in five Burkitt lymphoma cells lines. To test the role of HH signaling in Burkitt lymphoma, we exposed the cells to Sonic Hedgehog peptide or to SHH-containing conditioned media and measured the level of PTCH and GLI1 mRNA expression by quantitative RT PCR; cell viability by MTT assay; cell proliferation by BrdU incorporation; and apoptosis by caspase 3/7 assay. Although control cells (MC3T3) showed dramatic up-regulation of GLI1 and PTCH expression, the Burkitt lymphoma cell lines did not show changes in any of the assays. To inhibit HH signaling, we treated the Burkitt lymphoma cells with the SMO inhibitor, cyclopamine. Cyclopamine treatment did not affect GLI1 expression measured by quantitative RT PCR. Cyclopamine reduced viability of Burkitt lymphoma cells measured by MTT assays only when used at high concentrations (>5 µm), possibly representing a non-specific cytotoxic effect. Collectively, the results suggest that Burkitt lymphoma cells are not HH responsive and that constitutive c-MYC expression plays an important role in regulating GLI1 expression.

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 275. doi:10.1158/1538-7445.AM2011-275
Abstract 2754: Maternal use of antidepressants in pregnancy is associated with hypermethylation at the IGF2 imprinted control regions in the offspring in a race-dependent fashion
Authors: Soubry Adelheid , Murphy Susan K. , Huang Zhiqing , Murtha Amy , Schildkraut Joellen M. , Jirtle Randy L. , Wang Frances , Kurtzberg Joanne , Demark-Wahnefried Wendy , Forman Michele , Hoyo Catherine
Published: June 2012

In utero exposures to environmental factors may result in persistent epigenetic modifications affecting normal development and susceptibility to chronic diseases in later life. We determined whether exposure of the growing fetus to maternal depression or antidepressants is associated with aberrant DNA methylation at two differentially methylated regions (DMRs) of the imprinted Insulin-like Growth Factor 2 (IGF2) gene, namely the DMRs upstream of the imprinted IGF2 promoters (IGF2 DMR) and the DMRs upstream of H19 (H19 DMR). Aberrant DNA methylation at these DMRs has been associated with deregulated IGF2 expression, childhood cancers and several chronic diseases during adulthood. Our study population comprises pregnant mothers and their newborns (n=436), recruited between 2005 and 2008 in Durham, NC. A standardized questionnaire was completed and medical record data were abstracted to ascertain prenatal and periconceptional depression and antidepressive drug use. DMR methylation percentages in umbilical cord blood leukocytes were quantified using pyrosequencing. While an association between depression and methylation profiles was not found, we discovered significant hypermethylation of the H19 DMRs in newborns of African American -but not Caucasian- mothers who were taking antidepressive drugs during pregnancy (β=+6.89, p=0.01). Our findings suggest that changes in the IGF2/H19-domain methylation in response to maternal exposure to antidepressants are race-dependent.

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 2754. doi:10.1158/1538-7445.AM2011-2754
Abstract 2766: DNA methylation status of MLH1, p16 and SOCS1 in Puerto Rican patients with acute myeloid leukemia
Authors: Tirado-Gomez Maribel , Munoz Cristina , Cordero Paul R. , Bernabe Raul , Lacourt Mercedes , Rivera Noridza , Cotto Maribel
Published: June 2012

Background: DNA methylation of gene promoters have been show to play a role in the pathogenesis of acute myeloid leukemia. We present in this study a preliminary report of the methylation status of MLH1, p16 and SOCS1 in Puertorrican patients with acute myeloid leukemia

Methods: Peripheral blood was obtained from 24 patients with diagnosis of acute myeloid leukemia before their induction treatments. DNA was isolated and further modified with sodium bisulfite. Methylation-specific polymerase (MSP) chain reaction was performed to detect aberrant promoter methylation of MLH1, p16 and SOCS1.

Results: A total of 24 samples were analyzed. The mean age was 49 years (range 22-71) with 17/24 patients younger than 60 years (71%). Most of the patients were female (13/24, 54%). Six patients had diploid cytogenetics (25%), 6 had low risk cytogenetics (25%) and 12 had high risk cytogenetics (50%). Fifteen patients were de Novo AML (63%). Nine patients (38%) presented with WBC counts higher than 10 × 109/L. Thirteen patients (54%) achieved a complete response at the end of the induction therapy. Methylation of MLH1 was detected in 3/24 patients (13%). Methylation of p16 was detected in 5/24 patients (21%). Methylation of SOCS1 was detected in 2/24 patients (8%).

Conclusions: A tendency toward an association of methylated p16 with treatment resistance was noted. Also, methylation of SOCS1 was more common among relapsed AML patients. No correlation between methylated MLH1 gene and clinicopathological characteristics was noted. Collection of more patient's samples and correlation with clinical data and patient's ultimate outcome is ongoing.

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 2766. doi:10.1158/1538-7445.AM2011-2766
Abstract 2771: Genetic variation in interleukin, TLR, TNF and TRAF genes of NF-κB activation pathway and risk of prostate cancer
Authors: Liu-Mares Wen , Hu Jennifer J. , Konidari Ioanna , Allen Glenn O. , Clark Peter E. , Hall M. Craig , Dong Chuanhui , Rao J. Sunil , Scott William K.
Published: June 2012

It has been suggested that NF-κB triggers both the onset and resolution of inflammation by governing the expression of genes essential in activating malignancy-promoting signaling pathways. Members of the NF-κB activation pathway include tumor necrosis factor (TNF), several interleukins (IL), TNF receptor associated factors (TRAFs), and toll-like receptors (TLR). These molecules are involved in cell proliferation, differentiation, apoptosis and response to proinflammatory cytokines and mediating the production of cytokines necessary for the development of effective immunity. Common single nucleotide polymorphisms (SNP) in TNF, IL, TRAF, TLR and the main units of NF-κB genes were assessed for the association with prostate cancer (PrCA) risk in a clinic-based study of Caucasian and African-American cases and controls. A total of 709 tagSNPs (with minor allele frequency > = 5% and r2>=0.8) in 54 genes were genotyped in 903 Caucasians (552 cases and 351 controls) and 174 African-Americans (74 cases and 100 controls) using a custom Illumina Infinium II ® assay. Logistic regression models were used to examine each tagSNP with PrCA (adjusting for age and family history of prostate cancer and using an additive model for the SNP). Empirical p-values were generated by permutation (using 10,000 replicates) to correct for multiple comparisons. Statistically significant associations were observed in Caucasians at the NFkBIL1 and IL1R1 genes. The strongest evidence for association came at a polymorphism in the promoter region of the NFkBIL1 gene (rs3219184) (OR = 0.51 [95% CI 0.35-0.74], p = 0.0005), and a polymorphism in the 3’ flanking region of the IL1R1 gene (rs3917332) (OR = 1.60 [95% CI 1.23-2.09], p = 0.0005). We conclude that common variations in NFkBIL1 and IL1R1 might play important roles in PrCA susceptibility.

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 2771. doi:10.1158/1538-7445.AM2011-2771
Abstract 2782: Genetic susceptibility to prostate cancer progression and relationship with PSA level: Case-case study
Authors: Pashayan Nora , Neal David , Hamdy Freddie , Donovan Jenny , Martin Richard , Duffy Stephen , Pharoah Paul
Published: June 2012

Purpose: Prostate cancer is a mixture of progressive and non-progressive cancers. To date, genome-wide association studies have identified more than 30 common susceptibility variants associated with the risk of prostate cancer. However, case-control study designs have failed to identify variants consistently associated with aggressive cancer. The aim of this study is to investigate whether prostate cancer susceptibility variants distinguish between progressive and non-progressive cancers, using case-case study design.

Methods: We genotyped 85 prostate cancer susceptibility variants in 4,856 men with prostate cancer identified from the Prostate Testing for Cancer and Treatment (ProtecT), Prostate Cancer Mechanisms of Progression and Treatment (ProMPT), and Studies of Epidemiology and Risk Factors in Cancer Heredity (SEARCH) studies. Men with advanced stage (T3-T4) (n=624), or Gleason score ≥ 7 (n=1,792) prostate cancer were compared to those with localised stage (T1-T2) (n=3,805), or Gleason score <7 (n=2,090) cancer, respectively, using logistic regression to estimate the odds ratio (OR) associated with risk allele and genotype, after controlling for age at diagnosis, PSA level at diagnosis, stage and / or Gleason score.

Results: On univariate analysis, three variants (rs16901979, rs10896449, rs2735839) were associated with stage, and six (rs1447295, rs16901979, rs10896449, rs2735839 rs17181356 rs3760511) with Gleason score. In multivariate analysis, the variant rs2735839 at 19q13 in KLK2/3 region was associated with 42% increase in the odds for advanced stage cancer (adjusted OR per allele =1.42; 95% CI 1.08-1.87; ptrend =0.01). However, there was evidence of interaction with PSA level (p<0.001). The adjusted OR for rs2735839 for advanced compared to localised cancers among those with PSA ≥10ng/ml was 1.79 (95% CI 1.26-2.54; p=0.001), whereas among those with PSA<10ng/ml, the OR was 0.96 (95% CI 0.60-1.56; p=0.88). Whereas the variant rs3760511 at 17q12 was associated with reduced risk for high Gleason score cancer (adjusted OR per allele=0.85 95% CI 0.75-0.96; ptrend =0.01).

Conclusion: The interaction between PSA level and stage may inform the underlying biological mechanism for cancer progression in stage. These findings may also have clinical utility in identifying tumours with and without the potential for progression to advanced stage or higher Gleason score. Further validation in larger studies would be needed.

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 2782. doi:10.1158/1538-7445.AM2011-2782
Abstract 2784: Association between polymorphisms in angiogenesis-related genes and risk of urothelial carcinoma in Taiwan
Authors: Wang Yuan-Hung , Liu Chi-Tung , Yeh Shauh-Der , Wu Meei-Maan , Shen Kun-Hung , Shen Cheng-Hung , Chiou Hung-Yi
Published: June 2012

Introduction: Urothelial carcinoma (UC) including cancers of bladder, renal pelvis and ureter. Tumor staging and grading are considered to be useful indices for cancer prognosis. Recently, more and more studies indicated that angiogenesis is a necessary step in tumor growth and metastasis. This study aimed to investigate the relationship between single nucleotide polymorphisms (SNPs) of four angiogenesis-related genes including FIH-1, VHL, HIF-1 and VEGF and clinico-pathological characteristics, recurrence, and survival rate of UC.

Materials and Methods: In this case-control study, a total of 850 pathologically- confirmed UC cases and 850 cancer-free controls, frequency-matched on age, were recruited from those who admitted to the same hospitals for a health examination. Genotyping is determined using PCR-RFLP and Real-time PCR. Written informed consent was obtained from each of participants. This protocol was approved by the institutional review boards of the collaborated hospitals.

Results: We found that study subjects with cigarette smoking, alcohol consumption, occupational exposure, arsenic exposure, and high-salt diet have a significantly higher UC risk of 2.0, 1.6, 1.8, 1.9 and 1.5. Study subjects carrying C/G and G/G genotypes of FIH-1 gene had a 1.3-fold risk of developing to stage II or worse status. Cigarette smokers with the C/C genotype of VHL gene had a higher UC risk of 2.4. In addition, cigarette smokers with the T/T genotype of VEGF-460 C/T polymorphism had 1.6- fold UC risk of developing to stage II or worse status. In the combined analysis, study subjects who carried 2 or more risk genotypes of angiogenesis-related genes had a significantly higher UC risk of 1.6 for developing to T2 and stage II or more serious.

Conclusions: Our results suggested a significant association between polymorphisms in angiogenesis-related genes and morbidity and tumor stage of UC, however, without a significant association between these SNPs and tumor grade.

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 2784. doi:10.1158/1538-7445.AM2011-2784
Abstract 2788: Design, optimization, and biological evaluation of potent irreversible inhibitors of BTK kinase
Authors: Bearss David J. , Venkataswamy Sorna , Warner Steven L. , Vankayalapati Hariprasad , Bearss Jared J. , Sharma Sunil
Published: June 2012

Bruton's tyrosine kinase (BTK) is a cytoplasmic nonreceptor tyrosine kinase belonging to the Tec family of kinases. Critical for its function, BTK contains a pleckstrin homology (PH) domain and Src homology SH3 and SH2 domains. It signals downstream of the B-cell receptor (BCR) and is centrally involved in B-cell development. Activation of B-cells by various ligands is accompanied by the translocation of BTK to the cell membrane where it binds phosphatidylinositol-3,4,5-trisphosphate through its PH domain. Activation of BTK results in downstream signaling through the PI3K/AKT, PLCγ, NFκB, and other signaling pathways important for B-cell development and function. Recent reports have shown the aberrant expression and function of BTK in some cancers, including B-cell malignancies. We describe here the design, synthesis, molecular modeling, and biological evaluation of a series of small molecule, inhibitors of BTK kinase. Our initial lead compounds were identified via cross-docking experiments utilizing the crystal structure of BTK kinase and screening a previously in-house explored kinase inhibitor scaffold. We subsequently carried out structure-activity relationship studies and optimized the lead structures, which have IC50 activities in the range of 1 to 10μM against BTK. A critical step in the optimization of this chemical series against BTK was to explore the possibility of adding a Michael's acceptor group to react with Cys481 in the ATP-binding pocket of BTK. Optimization efforts yielded the currently best leads, HCI-1684 and HCI-1685, which inhibit BTK with IC50 values of 12 and 45 nM, respectively. Modeling data suggest these compounds irrepressibly bind in the ATP-binding pocket of BTK. HCI-1684, HCI-1685 and other lead compounds were further evaluated in cell-based assays and were demonstrated to inhibit BTK function downstream of BCR activation. This series of BTK inhibitors were shown to decrease phospho-PLCγ1/2 levels and other downstream phosphorylation events in malignant B-cell cell lines, such as Ramos B. Compound optimization and biological evaluation of this chemical series and evaluation in animal pharmacodynamic endpoint studies will be presented. Taken together, these results suggest BTK is a potential therapeutic target in cancer and that HCI-1684 is an exciting agent to potentially treat B-cell malignancies

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 2788. doi:10.1158/1538-7445.AM2011-2788
Abstract 2798: Tyrosine phosphorylation of LDH-A is important for cancer cell redox homeostasis and tumor growth
Authors: Fan Jun , Hitosugi Taro , Chung Tae-Wook , Wang Xu , Gu Ting-Lei , Roesel Johannes L. , Chen Zhuo (Georgia) , Boggon Titus , Khuri Fadlo R. , Kang Sumin , Chen Jing
Published: June 2012

Cancer cells take up more glucose than normal tissue and favor aerobic glycolysis, generating lactate through a NADH-dependent enzyme, lactate dehydrogenase A (LDH-A). This is the last step of glycolysis that permits the regeneration of NAD+, which is needed as an electron acceptor to maintain cytosolic glucose catabolism. Therefore, most tumor cells are reliant on lactate production for their survival. LDH-A gene expression is believed to be upregulated by both HIF and Myc in cancer cells to achieve increased lactate production, and expression of LDH-A was previously implicated to be involved in tumour initiation and growth. However, how oncogenic signals activate LDH-A to regulate cancer cell metabolism remains unclear. Our phospho-proteomics studies revealed that oncogenic fibroblast growth factor (FGF) receptor type 1 (FGFR1) tyrosine kinase directly phosphorylates LDH-A. Structural and biochemical studies revealed that phosphorylation at Y10 and Y83 activates LDH-A by promoting the formation of active, tetrameric LDH-A and binding of LDH-A substrate NADH, respectively. Moreover, we found that LDH-A is commonly phosphorylated at Y10 in diverse human cancer cells by multiple oncogenic tyrosine kinases including BCR-ABL, FLT3 and JAK2, which represents an acute molecular mechanism underlying increased lactate production in cancer cells. Furthermore, cancer cells with stable knockdown of endogenous LDH-A and rescue expression of a catalytic hypomorph LDH-A mutant, Y10F, show decreased cell proliferation and ATP levels under hypoxia, increased mitochondrial respiration to sustain glycolysis by providing NAD+, and reduced tumor growth in xenograft nude mice. Our findings suggest that tyrosine phosphorylation activates LDH-A to promote the Warburg effect and tumor growth by regulating NADH/NAD+ redox homeostasis in cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2798. doi:10.1158/1538-7445.AM2011-2798
Abstract 2806: Inactivation of PBRM1, a gene frequently mutated in clear cell renal carcinoma, suppresses tumor growth
Authors: Dachuan Huang , Kiat Ong Choon , Tean Teh Bin , Huimin Bernice Wong , Chan-on Waraporn , Subimerb Chutima , Furge Kyle , Futreal Andrew
Published: June 2012

The genetics of renal cancer is dominated by inactivation of the VHL tumour suppressor gene in clear cell renal carcinoma (ccRCC), the commonest histological subtype. However, our previous work has demonstrated that VHL loss alone is insufficient for ccRCC tumourigenesis arguing the need for additional genetic events. Recently, we have sequenced of the protein coding exome in 257 cases of primary renal carcinoma (RCC) and reported the identification of the SWI/SNF chromatin remodeling complex gene Polybromo 1 (PBRM1) as a second major RCC cancer gene with truncating mutations in 41% (85/257) of cases. In particular, all 9 cases with a SETD2 mutation have a mutation in either PBRM1 or VHL, with 7 of 9 cases having mutations in all three genes. The concomitant VHL, PBRM1 and SETD2 mutations, with all three genes mapping to chromosome 3p, suggest that the mutations are non-redundant functionally.

To obtain further evidence that PBRM1 can act as a cancer gene, transcriptional profiling before and after PBRM1 knockdown was performed using gene expression microarrays. The results showed that PBRM1 activity regulates pathways associated with chromosomal instability and cellular proliferation. Consistently, knockdown of PBRM1 RNA resulted in a significant increase in proliferation 4/5 RCC lines. No effect was seen, however, in A704 cell, which carries a homozygous truncating PBRM1 mutation, confirming the specificity of the assay. Further, knockdown of PBRM1 resulted in significantly increased colony formation in soft-agar and increased cell migration indicative of an increase in transformed phenotype. Taken together, these data support PBRM1 having a tumor suppressor role in ccRCC.

Therefore, understanding the contribution of PBRM1 mutation to clinical disease progression and outcome as well the potential for exploiting SWI/SNF complex abrogation therapeutically are important future areas of renal cancer research.

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 2806. doi:10.1158/1538-7445.AM2011-2806
Abstract 2808: Characterization of KRAS rearrangements in metastatic prostate cancer
Authors: Wang Xiao-Song , Shankar Sunita , Dhanasekaran Saravana M. , Ateeq Bushra , Sasaki Atsuo T. , Jing Xiaojun , Robinson Daniel , Cao Qi , Gopalan Anuradha , Reuter Victor E. , Cantley Lewis C. , Rubin Mark A. , Palanisamy Nallasivam , Varambally Sooryanarayana , Chinnaiyan Arul M.
Published: June 2012

In this study, we employed a novel integrative genomic-based approach to explore driving gene fusions contributing to the progression of advanced prostate cancer. This method, called amplification breakpoint ranking and assembly (ABRA), leverages the in vivo amplification and breakpoint analysis in cancer cells to assemble novel gene fusions and predict their tumorigenicity. Using ABRA, we nominated KRAS as a gene fusion with the ubiquitin-conjugating enzyme UBE2L3 in the DU145 cell line- which was originally derived from a metastatic prostate cancer to the brain. Interestingly, analysis of tissues revealed that 2 out of 62 metastatic prostate cancers harbored aberrations at the KRAS locus. In DU145 cells, UBE2L3-KRAS produces a chimerical protein which is readily ubiquitinated and relative instable, whereas specific knock-down of the fusion attenuates cell invasion and xenograft growth. Further, ectopic expression of the UBE2L3-KRAS fusion protein in NIH 3T3 fibroblasts and RWPE prostate epithelial cells exhibits substantial transforming activity in vitro and in vivo. In NIH 3T3 cells, UBE2L3-KRAS attenuates the MEK/ERK pathway, which is commonly engaged by oncogenic mutant KRAS. Instead, it enriches Ras proteins in late endosome, and diverts signaling to the AKT and p38 MAPK pathways. While a number of oncogenic activating point mutations of KRAS have been identified, this is the first description of a mutant chimeric version of KRAS that is oncogenic and thus may represent a new class of cancer-related alteration. This study also suggests that this aberration may drive metastatic progression in a subset of prostate cancers, which may be targeted by anti-KRAS therapies.

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 2808. doi:10.1158/1538-7445.AM2011-2808
Abstract 2817: An RNA aptamer is a potential biomarker for pancreatic cancer
Authors: Ray Partha , Sullenger Bruce A. , White Rebekah R.
Published: June 2012

Introduction: The symptoms of pancreatic cancer are usually non-specific and late. Existing tumor markers (such as CA19-9 and CEA) are neither sensitive nor specific enough to serve as useful screening tests. Tools for earlier detection could increase the proportion of patients who are cured with surgical resection.

Aptamers are oligonucleotide ligands that are identified from large libraries of RNA or DNA molecules using an iterative selection process known as SELEX (Systematic Evolution of Ligands by Exponential Enrichment). Aptamers bind their specific targets with high affinity and specificity and can be used for the dual purpose of identifying novel biomarkers and detecting them in biological specimens.

Methods: In order to select for aptamers against molecules that are selectively secreted by cancerous cells but not by normal cells, “positive/negative” SELEX was performed using the conditioned media (the “secretome”) of the MiaPaCa2 human pancreatic ductal adenocarcinoma cell line (positive selection) and the immortalized, non-cancerous human pancreatic ductal cell line HPDE (negative selection). Standard radioactive filter-based binding assays were used to measure binding.

Results/Conclusion: We have successfully selected several nuclease resistant (2′-fluoro-modified) RNA aptamers that differentially bind the secretome of pancreatic cancer cells compared to non-cancerous cells. One of the aptamers (M9-5) binds pancreatic cancer patient sera with high affinity compared to control sera collected from healthy donors. So far, the aptamer has demonstrated specific binding in 20 of 22 patients with histologically-proven pancreatic cancer and none of 20 healthy donors. In addition, paired serum samples obtained from two patients pre- and post-chemoradiation therapy and one patient pre- and two months post-resection have demonstrated a decrease in M9-5 binding with decreased tumor burden. Notably, the serum CA19-9 and CEA levels in several of these patients were normal, indicating that our biomarker might be more sensitive than these existing serum biomarkers for pancreatic cancer. M9-5 binding to the MiaPaCa2 “secretome” is sensitive to Proteinase K treatment indicating that the aptamer target is a protein. Currently, we are using a biotinylated version of the aptamer to affinity purify the target and identify it by mass-spectrometric techniques.

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 2817. doi:10.1158/1538-7445.AM2011-2817
Abstract 2818: Urinary MMP-2 and MMP-9 predict the presence of ovarian cancer in women with normal CA125 levels
Authors: Coticchia Christine M. , Curatolo Adam S. , Zurakowski David , Matulonis Ursula A. , Moses Marsha A.
Published: June 2012

Currently, the only widely used biomarker for monitoring ovarian cancer status and response to therapy is the cancer antigen 125 (CA125). Interestingly, the majority of newly developed ovarian cancer multi-marker assays also rely heavily on serum CA125 levels as one of the biomarkers in their panels. However, 20% of women with advanced ovarian cancer have blood CA125 levels within the normal range of 35 U/mL and below. These women represent a cohort of patients for whom no clinically useful biomarkers are available for monitoring disease status, stage or response to therapy. To address this need, we asked whether a panel of urinary biomarkers that we have previously validated for use in other cancers, could predict the presence of disease in women with ovarian cancer possessing normal CA125 levels. Urine samples were collected from 81 healthy controls and 26 ovarian cancer patients with blood CA125 levels below 35 U/mL. Using monospecific ELISAs, we determined that urinary levels of matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) and the neutrophil gelatinase lipocalin 2 (NGAL) are detected at significantly higher levels in the urine of ovarian cancer patients than in the urine of healthy controls. Additionally, receiver-operating characteristic (ROC) area under the curve (AUC) analysis of these groups revealed that, individually, MMP-2, MMP-9 and NGAL each could significantly distinguish ovarian cancer patients with CA125 < 35 U/mL from controls. The best combination of biomarkers for predicting the presence of disease was MMP-2 plus MMP-9 with an ROC AUC of 0.7332 (P<0.01). When these biomarkers were multiplexed with age, a know risk factor of ovarian cancer, the diagnostic accuracy of these biomarkers increased to an AUC of 0.820 (P<0.001). This combination of urinary MMP-2, MMP-9 and age demonstrated excellent diagnostic capability in differentiating ovarian cancer patients from healthy controls among the cohort of patients for whom CA125 values were uninformative.

Acknowledgments: The authors gratefully acknowledge the support of Ovations for the Cure of Ovarian Cancer (MAM and UAM) and the Stuart Weitzman Foundation (MAM).

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 2818. doi:10.1158/1538-7445.AM2011-2818
Abstract 2841: Inducible modulation of NF-kappaB in macrophages reveals that timing of intervention may be critical to therapeutic outcome during mammary tumorigenesis
Authors: Barham Whitney , Connelly Linda , Onishko Halina M. , Chen Lianyi , Sherrill Taylor , Zabuawala Tahera , Ostrowski Michael , Blackwell Timothy , Yull Fiona
Published: June 2012

Elevated levels of NF-kappaB signaling are associated with multiple cancers. Therefore, it is not surprising that inhibitors of NF-kappaB are being investigated as potential therapeutics. While extensive efforts are being made to identify inhibitors there is less focus on investigating the effects of inhibition in specific cell types and during defined stages of tumor progression. We have developed murine transgenic models that enable us to induce expression of an activator or inhibitor of NF-kappaB in macrophages, by adding doxycycline to the drinking water of mice. We have combined these with the Polyoma model of mammary cancer to investigate the effects of modulation of NF-kappaB signaling specifically within macrophages. Given several recent publications we were predicting that activation of NF-kappaB would have pro-tumor effects. However, using a tail vein injection model, our data identifies a stage during progression in which activation of NF-kappaB in macrophages significantly limits metastasis. Activation of NF-kappaB alters immune cell populations and results in rapid killing of tumor cells during the seeding phase. In this model inhibition of NF-kappaB in macrophages during this critical stage of metastasis is pro-tumor. This suggests that while inhibition of NF-kappaB at later stages of tumor progression may have therapeutic benefit, inhibition during the seeding phase my lead to the opposite effect. Our studies highlight the importance of continued investigation into aspects of cell specificity and timing that may change the potential outcome of a clinical trial using NF-kappaB inhibitors.

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 2841. doi:10.1158/1538-7445.AM2011-2841
Abstract 2846: Pigment epithelium-derived factor (PEDF) functions as a brain metastasis suppressor of breast cancer
Authors: Fitzgerald Daniel P. , Palmieri Diane , Qian Yongzhen , Deshpande Monika , Subramanian Pretti , Graves Christian , Davis Sean , Meltzer Paul , Camphausen Kevin , Becerra Patricia S. , Steeg Patricia S.
Published: June 2012

Brain metastases occur in 10-20% of metastatic breast cancer patients. The one year survival rate is approximately 20%. Brain metastasis is typically diagnosed late in breast cancer progression; however, studies suggest that the incidence of brain metastasis may be increasing under current chemotherapeutic regimens. We have investigated changes in gene expression associated with the acquisition of brain metastatic potential by breast cancer. A DNA microarray analysis comparing human brain metastases and a cohort of unlinked primary breast carcinomas identified Pigment Epithelium-Derived Factor (PEDF) as down-regulated in the brain metastases relative to the primary tumors (mean ∼14X lower by QPCR validation). The data suggest the hypothesis that PEDF may have a negative impact on brain metastatic progression. PEDF, a secreted factor, has been shown to act as a tumor suppressor, has strong anti-angiogenic activity, but supports the proliferation/viability/ differentiation of neural tissue. We hypothesize that the multiple roles of PEDF make it a potent agent which targets not only tumor cells, but also the microenvironment of the brain. In vitro experiments demonstrated that PEDF can promote death of breast cancer cells. Conversely, PEDF promoted neuronal survival, in vitro. In a xenograft model system (231BR, a brain-tropic derivative of human MDA-MB-231 breast cancer cells) PEDF overexpression inhibited formation of large brain metastases (∼2-fold, p=0.001) and reduced proliferation of PEDF-expressing breast cancer cells in mouse brain metastases (11% Ki67-positive, PEDF-positive vs. >56% Ki67-positive, PEDF-negative). In an intracranial implantation model, expression of PEDF by breast cancer cells lead to a rapid reduction in brain metastatic tumor volume. Histological examination suggests that PEDF reduces gliosis and protects neurons concomitant to suppression of metastatic growth. The data suggest the intriguing possibility that PEDF overexpression may concurrently inhibit brain metastasis of breast cancer cells but preserve neuronal function.

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 2846. doi:10.1158/1538-7445.AM2011-2846
Abstract 2865: Cytoplasmic deacetylase HDAC6 confers tumor suppressive potential through autophagic cell death in human hepatocellular carcinoma
Authors: Jung Kwang Hwa , Noh Ji Heon , Eun Jung Woo , Kim Jeong Kyu , Bae Hyun Jin , Xie Hongjian , Nam Eliel , Park Won Sang , Lee Jung Young , Nam Suk Woo
Published: June 2012

The histone modification and acetylation by histone acetyltransferases (HATs) and histone deacetylases (HDACs) have been associated with chromatin decondensation and transcriptional activation and silencing. HDAC6, one of class II HDAC isotypes, is a cytosolic protein and microtubule-associated deacetylase that interacts with non-histone protein, including α-tubulin. HDAC6 is involved in autophagic degradation of the cells, but the underlying mechanism associated with carcinogenesis in solid tumors remains unclear. In this study, we investigated the expression levels of HDAC6 in human liver cancer cell lines and tissues. HDAC6 expression was analyzed by Western blot analysis in nine selected hepatocellular carcinomas (HCCs) and immunohistochemical staining was performed in paraffin-embedded samples from 32 liver cancers. HDAC6 expression was significantly down-regulated in the HCCs compared to corresponding normal tissues in nine selected tissue samples. Immunohistochemical analysis also showed commonly down-regulated HDAC6 expression in the HCCs. Thus, to investigate the HDAC6-induced anti-neoplastic potentials on liver cancer progression, we restored HDAC6 expression using gene transfection in human Hep3B liver cancer cell line. Over-expression of HDAC6 suppressed proliferation of liver cancer cells with decreased acetylation status of α-tubulin and exhibited the characteristics of autophagic degradation (autophagosome formation and the LC3-II generation) in Western blot and Electron Microscopy (EM) analysis. In addition, we found that over-expression of HDAC6 induced expression of autophagy-related gene Beclin 1, a key molecule of autophagosome formation. In contrast, knock-down of HDAC6 over-expressed cells could reduce the expression level of increased Beclin 1 and LC3-II with restoration of acetylated α-tubulin. Furthermore, in the xenograft mouse models, over-expression of HDAC6 significantly suppressed tumor growth and incidence of tumor formation. In conclusion, our results suggest that HDAC6-indued autophagic cell death provide a novel mechanism for the regulation of beclin1 and LC3 expression in liver cancer development and progression providing new therapeutic intervention in human hepatocellular carcinoma

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 2865. doi:10.1158/1538-7445.AM2011-2865
Abstract 287: A SILAC proteomics analysis to identify bladder cancer metastasis-associated candidates
Authors: Grau Laura , Luque-García Jose L. , González-Peramato Pilar , Theodorescu Dan , Gil Marta , Palou Joan , Bellmunt Joaquin , Sanchez-Carbayo Marta
Published: June 2012

Bladder cancer is most clinically aggressive when metastases develop, associated with the shortest patient survival. To identify molecular pathways associated with the metastatic process in bladder cancer, we carried out a proteomic analysis of an invasive T24 and a metastatic derived T24T bladder cancer cell line using a SILAC (Stable Isotope Labelling by Amino acids in Cell culture) approach. Protein extracts were fractionated, separated by SDS-PAGE and analyzed by nanoflow LC-ESI-LTQ mass spectrometry (MS). Comparative analysis of the invasive and metastatic bladder cancer cells identified 831 proteins with high confidence (>99%). Protein expression data from SILAC were compared with mRNA expression data obtained from oligonucleotide microarrays. Relatively weak correlations were observed, consistent with the poor associations between RNA and protein expression previously reported. The involvement of the proteins identified in biological networks served to propose how selected candidates may participate in molecular pathways associated with bladder cancer metastasis. Verification analyses included Western Blot using independent protein extracts. Immunohistochemistry analyses of proteins differentially expressed, such as Cul3, a protein involved in the ubiquitination and subsequent proteasomal degradation of target proteins, were performed on bladder tumours spotted onto tissue microarrays (n=284). Cul3 was associated with tumor staging, lymph node metastasis and disease-specific survival. Further functional analyses revealed that Cul3 silencing diminished the proliferative, migration and invasive rates of T24T cells, and restored the expression of other metastasis related cytoskeleton proteins identified by SILAC such as ezrin, moesin, filamin A or caveolin. Hence, this approach not only identified proteins and molecular mechanisms involved in bladder metastasis but also revealed a role for Cul3 in bladder cancer progression, metastasis and clinical outcome.

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 287. doi:10.1158/1538-7445.AM2011-287
Abstract 2870: The inhibitory effects of propolin C and propolin G on human lung adenocarcinoma A549 cells through autophagy and apoptosis pathway
Authors: Hsu Hui-Chen , Ku Feng-Chuan , Chen Yu-Fen , Wang Hsiao-Feng , Lin Zhong-Zhe , Chen Yue-Wen , Tsai Meng-Geng
Published: June 2012

Propolis has been used empirically for centuries and it was always mentioned as an immunomodulatory agent. Propolis has over 150 constituents such as polyphenols (flavonoids, phenolic acids and their esters), terpenoids, steroids, and amino acids, but its composition varies qualitatively and quantitatively with the geographical and botanical origins. In Tawiwan, there are ten propolins (A-J) were isolated and characterized from the local propolins. Propolins, prenylflavanones are isolated and characterized from Taiwanese propolis have been shown inhibits certain cancer cell lines growth and induced apoptosis. In this study, human lung adenocarcinoma cell (A549) was used to investigate the anticancer activity of Proploins. Propolin C and propolin G could inhibit A549 cell growth by dose-dependent manner. The combination of propolin C with propolin G was more predominantly inhibited the cell viability. Propolin C or propolin G treatment alone could suppress Akt and p70S6K activation by blocking the PI3K/Akt/mTOR pathway. Moreover, the pro-apoptotic protein Bax and Bad expression were increased. The procaspase 9, procaspase 3, and PARP activated in cascade to lead to mitochondria-mediated apoptosis pathway of apoptosis. Flow cytometry assay showed that sub-G1 phase increased significantly in propolin G treatment but not in propolin C group. Both propolin C and G could promote LC3B increased within 6 hours treatment, moreover, and AVOs (acidic vesicular organelles) significantly increased by AO (Acridine orange) staining. Combination treatment had synergies effect to induce significantly apoptosis and autophagy in A549 cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2870. doi:10.1158/1538-7445.AM2011-2870