bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3938: Altered nucleotide excision repair gene expression after cisplatin treatment in melanoma
Authors: Bowden Nikola A. , Ashton Katie A. , Avery-Kiejda Kelly A. , Zhang Xu Dong , Hersey Peter , Scott Rodney J.
Published: February 2011

Melanoma incidence rates in Australia are the highest in the world. Two of the hallmark features of melanoma are the development of disease as a result of chronic repeated ultraviolet (UV) radiation exposure and the limited efficacy of cisplatin in treatment. These features have one commonality; both result in large helix distorting DNA damage that is recognised and repaired by the DNA repair system, nucleotide excision repair (NER). The aim of this study was to examine the expression of NER gene transcripts in melanoma cell lines treated with cisplatin.

One melanocyte, three primary melanoma (MM200, IgR3, Me4405) and two metastatic melanoma (Mel-RM and Sk-Mel-28) cell lines were used for this study. The cells were treated with cisplatin (10μg/mL) and RNA was harvested at 0, 6 and 24 hours after treatment. Thirteen NER gene transcripts were measured by semi-quantitative real-time PCR. The relative expression of each gene was calculated and t-tests were used to identify significantly altered expression in melanoma compared to melanocytes and between timepoints.

The genes involved in the global genome repair (GGR) DNA damage recognition pathway; XPC, DDB1 and DDB2 (XPE) showed significantly decreased relative expression (RE) in the melanomas compared to the melanocytes, 24 hours after cisplatin treatment. In contrast, the genes involved in the transcription coupled repair (TCR) DNA damage recognition pathway, ERCC8 (CSA) and ERCC6 (CSB) showed low RE in all cell lines at all time points. Furthermore, the genes involved in the convergent NER pathway revealed no significant change in RE 24 hours after cisplatin treatment in the melanomas, however, an increase in RE was seen in the melanocytes. Overall, the basal expression levels of all genes investigated were greater in the melanoma cell lines compared to the melanocytes.

The GGR DNA damage recognition arm of the NER pathway is essential to maintain genomic stability through the identification of helix distorting DNA adducts arising from UV radiation and genotoxic chemicals. The findings from this study revealed reduced GGR transcript levels 24 hours after cisplatin treatment. This finding may be the first step towards identifying the biological mechanisms underlying the limited efficacy of cisplatin in melanoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3938.
Abstract 3944: Base excision repair and gene expression profiling in malignant melanoma
Authors: Ashton Katie A. , Bowden Nikola A. , Kairupan Carla F. , Avery-Kiejda Kelly A. , Zhang Xu Dong , Hersey Peter , Scott Rodney J.
Published: February 2011

Melanoma is a potentially deadly skin tumour and its incidence continues to increase worldwide. Despite the many efforts to elucidate the molecular mechanisms involved in melanoma development, these pathways still remain relatively unknown. For this reason, new methods are required to identify new biological targets implicated in melanoma. Our pilot study examined whole genome gene expression (Illumina Ref8 V2) in 82 melanoma samples and 8 control melanocyte cell lines. Gene transcripts with significantly altered expression (p<0.05) and greater than 2-fold change were identified for the melanomas compared to the melanocytes (GeneSpring GX V10), revealing a number of new targets not previously described in melanoma. Interestingly, a number of genes involved in the DNA repair, immune response and cell cycle arrest pathways were differentially expressed in the melanomas. With respect to the DNA repair pathways, we identified that key components of the base excision repair (BER) pathway were significantly altered in melanoma. Sequencing analysis confirmed several alterations in genes involved in the BER pathway and the frequency of these variants were further confirmed in an independent cohort of 105 formalin-fixed paraffin embedded (FFPE) melanoma patients. Hierarchical cluster analysis showed that melanoma patients with sequence variants in the BER pathway had altered gene expression profiles compared to those melanoma patients with no known variants in this pathway. Taken together, these results suggest that the BER pathway may be implicated in the development of melanoma, which may lead to the introduction of mutation screening and gene expression profiling for the management of melanoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3944.
Abstract 3953: C-Kit and Mitf signaling: Novel involvement of the PI3K/Akt and p38 pathways
Authors: Phung Bengt , Schepsky Alexander , Steingrimsson Eirikur , Rönnstrand Lars
Published: February 2011

The development of melanocyte from melanoblast is regulated by the tyrosine kinase receptor c-Kit and the basic-helix-loop-heliz-leucine zipper transcription factor Mitf. It is established that Mitf activity is controlled by c-Kit activation and subsequent phosphorylation of the Erk/Mapk pathway. Serine phosphorylation (activation) of Mitf not only leads to upregulation of Mitf target gene expression but also ubiquitin dependent proteosomal degradation of Mitf. There is little knowledge surrounding the dysregulation between c-Kit and Mitf signaling that is often found in melanocyte progression to melanoma. The aim of our study is, therefore, to identify the signal transduction pathways involved in c-Kit mediated regulation of Mitf.

In our study, we found c-Kit tyrosine phosphorylation sites and novel signaling pathways involved in the regulation of Mitf. Phosphorylation of c-Kit at Tyr721 has been shown to activate the PI3K pathway and Tyr568 and Tyr568/570 are responsible for Src kinase mediated activation of Erk/Mapk pathway. We demonstrate that Y721F, Y568F and Y568/570F mutation of c-Kit prevented Mitf mobility shift, indicating that these tyrosine phosphorylation sites are involved in Mitf regulation. Further, we observe that treatment of c-Kit and Mitf transfected HEK293T cells with LY294002 (PI3K inhib.), Akt IV (Akt inhib.), SU6656 (Src kinase inhib.), SB203580 (p38 inhib.) or U0126 (Mek inhib.) prevented c-Kit mediated Mitf mobility shift. Similar results were also observed in the murine melanocyte, Melan- A cell line where Mitf and c-Kit are endogenously expressed. In order to further verify c-Kit's role as Mitf regulator, we examined tyrosinase (downstream target of Mitf) promoter activity using a luciferase reporter construct. Ligand stimulation of c-Kit increased the Mitf target promoter activity.

In conclusion, for the first time, we demonstrate that the signal transduction between c-Kit and Mitf is dependent on, 1) c-Kit Tyr721/PI3K/Akt pathway, 2) p38 pathway and 3) c-Kit Tyr568 or Tyr568/570 / Src kinase mediated Erk pathway. C-Kit activation leading to downstream were also shown to promote the transcriptional activity of Mitf. Our results reveal novel mechanisms by which c-Kit signaling regulates Mitf, with implication for understanding of both melanocyte development and melanoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3953.
Abstract 3966: 14-3-3 sigma and 14-3-3 zeta plays an opposite role in cell growth inhibition mediated by transforming growth factor-beta 1
Authors: Hong Hye-Young , Jeon Woo-Kwang , Park Sung-Ji , Jang Hyung-Ju , Choi Ji-Yeon , Kim Byung-Chul
Published: February 2011

The expression of 14-3-3 proteins is dysregulated in various types of cancer. This study was undertaken to investigate the effects of 14-3-3 ζ and 14-3-3 σ on cell growth inhibition mediated by transforming growth factor-beta 1 (TGF-β1). Mouse mammary epithelial cells (Eph4) that are transformed with oncogenic c-H-Ras (EpRas) and no longer sensitive to TGF-β1-mediated growth inhibition displayed increased expression of 14-3-3 ζ and decreased expression of 14-3-3 σ compared with parental Eph4 cells. Using small interfering RNA-mediated knockdown and overexpression of 14-3-3 σ or 14-3-3 ζ, we showed that 14-3-3 σ is required for TGF-β1-mediated growth inhibition whereas 14-3-3 ζ negatively modulates this growth inhibitory response. Notably, overexpression of 14-3-3 ζ increased the level of Smad3 protein that is phosphorylated at linker regions and cannot mediate the TGF-β1 growth inhibitory response. Consistent with this finding, mutation of the 14-3-3 ζ phosphorylation sites in Smad3 markedly reduced the 14-3-3 ζ-mediated inhibition of TGF-β1-induced p15 promoter-reporter activity and cell cycle arrest, suggesting that these residues are critical targets of 14-3-3 ζ in the suppression of TGF-β1-mediated growth. Taken together, our findings indicate that dysregulation of 14-3-3 σ or 14-3-3 ζ contributes to TGF-β1 resistance in cancer cells.

Following are results of a study on the “Human Resource Development Center for Economic Region Leading Industry” Project, supported by the Ministry of Education, Science & Technology(MEST) and the National Research Foundation of Korea(NRF)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3966.
Abstract 3972: The role of the TGF-β type III receptor in colon carcinogenesis
Authors: Holtzhausen Alisha , Gatza Catherine E. , Kirkbride Kellye C.
Published: February 2011

The loss of the TGF-β type III receptor (TβRIII), a TGF-β superfamily co-receptor, correlates with disease progression in multiple cancer types, suggesting that TβRIII plays a role in regulating tumor progression. TGF-β superfamily signaling is frequently dysregulated in colon cancer but the specific role of TβRIII in colon carcinogenesis has not been examined. In contrast to other cancer types, TβRIII mRNA expression is not altered in colon cancer and there is not a significant change in TβRIII protein expression in tumors compared to normal colon tissue. HT29 colon cancer cells overexpressing TβRIII (HT29-RIII) exhibit an increase in proliferation in response to TGF-β and BMP2 stimulation compared to HT29-Neo cells. This increase in proliferation is mediated through the down-regulation of p21. In a soft agar assay HT29-RIII cells demonstrate increased colony formation in response to ligand treatment. Additionally, HT29-RIII cells are resistant to anoikis and 5FU-induced apoptosis. Colon cancer cells expressing TβRIII also exhibit increased migration potential and this increase in migration can be abrogated by knockdown of TβRIII expression. Furthermore, cells over-expressing TβRIII show alterations in E-cadherin and actin organization. Taken together, these data suggest that TβRIII may have a pro-tumorigenic role in colon cancer tumorigenesis as it mediates ligand stimulated proliferation, resistance to apoptosis and increased migration. Further work is currently underway to more fully elucidate the role of TβRIII in colon cancer progression in vitro and in vivo.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3972.
Abstract 3977: Gene-expression signature specifically associated with triple-negative breast cancer pertaining to asian ethnicity
Authors: Chang Yao-Yin , Kuo Yu-Ho , Chen Hung Lee , Lai Liang-Chuan , Kuo Wen-Hung , Hsiao Chuhsing Kate , Chuang Eric Y. , Chang King-Jen
Published: February 2011

Triple-negative breast cancer is a subtype of breast cancer with strong aggressiveness of tumor behavior and distinct patterns of metastasis. This unique phenotype of breast cancer is given by its clinically negative expression of estrogen receptor (ER), progesterone receptors (PR), and HER2 protein. Triple-negative breast cancer patients often suffer from ineffective hormone therapies, high recurrence rate, and a predilection for visceral metastasis. An accurate gene-expression signature specifically associated with triple-negative breast cancer will facilitate the identification of the existence of this disease and provide insights of its etiology.

DNA microarray analysis was used to evaluate gene-expression profiles of 181 Asian breast cancer patients. There were 48 breast cancer patients who were triple-negative, and 133 of them were non-triple-negative. Several clinical information were matched in each group, including lymphovascular invasion, age, stage, grade, tumor size, tubule formation, nuclear pleomorphism, and mitotic count. After all 181 microarray data were normalized with quantile normalization, significantly expressed genes differentiated from the triple-negative group and non-triple-negative group were extracted using SAM (Significant Analysis of Microarray) and T-test. A panel of 50 genes was selected as the gene-expression signature for triple-negative breast cancer for Asian ethnicity, based on their unique patterns of gene-expression data using hierarchical clustering. Pathway analysis of the signature genes for triple-negative breast cancer was then performed in the lab. Genetic interactions of significantly expressed genes among different pathways found in this study will be shown and elaborated on the poster.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3977.
Abstract 3993: Characterisation of early ovarian cancer biomarkers
Authors: Meehan Katie L. , Rainczuk Adam , Fairweather Nicole , Stephens Andrew
Published: February 2011

Around 22,000 new cases and 15,500 deaths from ovarian cancer are expected in the US in 2008, and currently women run an estimated 1/67 risk of developing ovarian cancer during their lifetime. Approximately 70% of patients present with late stage disease (stages III - IV; FIGO classification) and have a poor prognosis, with only a ∼30% 5 year survival rate. By contrast, patients diagnosed with early (stage I) disease exhibit survival rates above 90%, with disease often cured by surgery alone. The key issue is that early detection is hampered by a lack of sensitive and specific markers. The aim of this ongoing study was to apply 8-plex isobaric tags for relative and absolute quantitation (iTRAQ) for the proteomic investigation for early stage markers of ovarian cancer.

Plasma was collected from patients and pooled as follows: G1-early stage cancers (stage I/II), G2-late stage cancers (stage III), G3-ascites fluid, G4-benign adenomas, G5-benign adenofibromas, G6-controls and G7-pool of all samples. Pooled patient samples were immuno-depleted of 12 abundant, disease-unrelated proteins and concentrated using reverse phase chromatography. Equal protein concentrations of each of the 7 pools were reduced, denatured, alkylated and digested using trypsin. Peptides from G1 through G7 were labelled with iTRAQ labels −113 to −119 respectively. Samples were combined and analysed using two-dimensional liquid chromatography (2D-LC) and mass spectrometry (4700 MALDI-TOF/TOF or QStar Elite, Applied Biosystems). Data were analysed using Protein Pilot (version 3.0, Applied Biosystems).

Initial studies have identified 111 proteins with quantitation of 71 proteins, including numerous proteins that have been patented as candidate early markers for ovarian cancer. Of those identified, many are involved in inflammation/immunity and coagulation. Of considerable interest was the quantitative identification of numerous low abundance proteins not previously associated with early stage ovarian cancer.

This proteome analysis has generated a catalogue of proteins that display potential as early markers for ovarian cancer. In addition, this study will assist in our understanding of the pathways potentially involved in the onset and progression of ovarian cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3993.
Abstract 3999: Human proteome arrays for auto-antibody identification in clinical cancer studies
Authors: Kenten John H. , Suschak Jessica , Roth Kristian , Jeffery-Coker Bandele , Yin Fei , Glezer Eli N.
Published: February 2011

Early detection, identification and classification of cancers still present considerable challenges. Innovations in several areas of research have provided novel and powerful approaches with which to tackle these challenges. Autoantibodies to tumor antigens have been found in cancer patients. Such autoantibodies are potentially ideal diagnostic biomarkers, as they are present in serum, are stable, and can be measured using established serological techniques. In order to discover new autoantibody responses in cancer patients, we have developed an approach based on the generation of protein arrays for systematic identification of auto-reactive antibodies. Our approach makes use of several key technologies: sequence-curated clone libraries, in-vitro protein expression, and a sensitive detection technology based on a proprietary combination of electrochemiluminescence detection and patterned arrays (Multi-Array® Technology, Meso Scale Diagnostics). Using this combination of technologies we plan to generate immobilized arrays of >1,000 proteins, with a high density of proteins per well in a 96-well microtiter plate format. The array of human proteins is selected from our collection of 60,000 mammalian clones (characterized by sequencing). The protein arrays will be used to screen cancer patient sera and controls.

During the initial development of this platform we optimized the protocols for in-vitro protein expression and labeling, specifically for immobilization on our Multi-Array plates. With this approach we have demonstrated detection of antigen-specific antibodies down to a concentration of 0.1 ng/mL. We have also validated this approach in a model system, with the detection of auto-reactive antibodies in the sera from 29 patients with a range of auto-immune diseases (Sjogren's syndrome, mixed connective tissue disease, scleroderma, systemic lupus erythematosus, and primary bilary cirrhosis), using 9 cloned autoimmune antigens. In parallel, we also optimized approaches for spotting in-vitro expressed proteins in high density spot/well arrays. In these studies we evaluated a selection of electrode surfaces and protein coupling methods, in combination with various spotting methods, and demonstrated serological measurements in a high density spot/well array. This effort was supported with funding from the NCI #1R44CA130391-1A1.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3999.
Abstract 4006: Soluble ALK4 extracellular domain antagonizes oncogenic Cripto signaling and inhibits tumor growth
Authors: Spike Benjamin T. , Kelber Jonathan A. , Booker Evan C. , Klemke Richard L. , Wahl Geoffrey M. , Vale Wylie W. , Gray Peter C.
Published: February 2011

Cripto is a small GPI-anchored glycoprotein that plays key roles during normal development and in tumorigenesis. Cripto is expressed at aberrantly high levels in human tumors and blocking its oncogenic effects on activin/Nodal/TGF-beta, Erk/MAPK and PI3K/Akt pathways represents a promising therapeutic strategy. Cripto functions as a Nodal co-receptor and binds directly to the extracellular domain (ECD) of the activin/Nodal type I receptor ALK4. We have shown that a kinase-deleted ALK4 mutant deficient in activin binding (ALK4-L75A) retains Cripto binding and can block Cripto effects on activin and Nodal signaling. We hypothesized that soluble ALK4-L75A ECD proteins would similarly bind Cripto and disrupt oncogenic Cripto function. To test this, we generated two soluble constructs incorporating the ALK4-L75A ECD, one with a C-terminal His tag (sALK4-L75A-His) and the other consisting of an ALK4-L75A ECD-Fc fusion with an N-terminal FLAG tag (sALK4-L75A-Fc). We expressed the sALK4-L75A-His and sALK4-L75A-Fc proteins in 293T cells and then purified them from conditioned media using Ni-NTA agarose and Protein-A agarose, respectively. The sALK4-L75A-Fc protein dose-dependently inhibited 125I-Cripto binding to 293T cells transfected with full length ALK4. In addition, sALK4-L75A-Fc blocked 125I-Cripto binding to untransfected human mammary epithelial MCF10A cells. In accordance with these binding data, sALK4-L75A-His inhibited Cripto-dependent Nodal signaling in 293T cells and sALK4-L75A-Fc inhibited Nodal-induced Smad2 phosphorylation in human NCCIT cells. sALK4-L75A-His also blocked Cripto-induced PI3K signaling while sALK4-L75A-Fc inhibited Cripto-induced MCF10A cell proliferation. Importantly, we have shown that sALK4-L75A-Fc can also inhibit the growth of tumor xenografts derived from NCCIT cells at doses where no toxicity was observed. Thus, we provide the first evidence that soluble forms of ALK4 inhibit oncogenic Cripto signaling. The efficacy of a soluble ALK4 receptor as an antitumor agent points to the utility of Cripto antagonists in the treatment of human cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4006.
Abstract 4007: Cooperative regulation of Stat3 signaling by TC-PTP, SHP1, and SHP2 in keratinocytes following UVB irradiation
Authors: Kim Dae Joon , Tremblay Michel L. , DiGiovanni John
Published: February 2011

Stat3 is initially dephosphorylated in murine keratinocytes in response to UVB irradiation. Treatment with Na3VO4 desensitized keratinocytes to UVB-induced apoptosis with the recovery of phosphorylated Stat3 protein levels, implying that a protein tyrosine phosphatase (PTP) is involved in this mechanism. In the current work, we report that three PTPs including TC45, the nuclear form of TC-PTP, SHP1, and SHP2 are involved in this rapid dephosphorylation of Stat3 in keratinocytes induced by UVB irradiation. Dephosphorylation of Stat3 was increased rapidly after UVB irradiation of cultured keratinocytes. Knockdown of TC-PTP, SHP1, or SHP2 using RNAi showed that these PTPs are in part responsible for Stat3 dephosphorylation following UVB irradiation. The level of phosphorylated Stat3 was significantly higher in keratinocytes transfected with TC-PTP, SHP1, or SHP2 siRNA in the presence or absence of UVB compared with keratinocytes transfected with control siRNA. TC45, the isoform of TC-PTP expressed in keratinocytes, was mainly localized in the cytoplasm of keratinocytes and translocated from cytoplasm to nucleus upon UVB irradiation. Stat3 dephosphorylation was associated with nuclear translocation of TC45. Further studies revealed that simultaneous knockdown of all three phosphatases, TC45, SHP1, and SHP2, using RNAi recovered the level of phosphorylated Stat3 upon UVB irradiation. In mouse epidermis, the level of phosphorylated Stat3 also was initially decreased by UVB irradiation followed by a significant increase at later time points after UVB exposure. The expression patterns of Stat3 target genes, such as cyclin D1 and c-Myc, followed the changes in activated Stat3 in response to UVB irradiation. Collectively, these results suggest that three phosphatases, TC45, SHP1, and SHP2, are responsible for UVB-mediated Stat3 dephosphorylation and may serve as part of an initial protective mechanism against UV skin carcinogenesis. Supported by NIH grants CA105345, CA76520 and an Odyssey Fellowship to DJK.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4007.
Abstract 4016: Large scale, domain-based comparative protein interaction mapping in cancer cell signaling
Authors: Hindie Valerie , Tafelmeyer Petra , Meil Alain , Collura Vincent , Hamburger Alexandre , Wojcik Jérôme , Barillot Emmanuel , Boissy Guillaume , Camonis Jacques , Daviet Laurent , Formstecher Etienne
Published: February 2011

Protein interaction mapping has proven instrumental for the delineation and understanding of cancer signaling pathways, both in Homo sapiens and model organisms. We have recently published a drosophila protein interaction map centered on fly orthologs of human cancer-related and signaling proteins1.

We now report the completion of the cognate human interaction map using the same high-throughput, domain-based yeast two-hybrid (Y2H) technology. 149 human homologs of the 102 drosophila proteins, including 29 human oncogenes and tumor suppressors2, were used as entry points to screen a highly complex, random-primed placenta cDNA library comprised of 10 million independent fragments in yeast. In order to strictly parallel the drosophila study, the same domains were selected as baits on human orthologs, resulting in 230 different screens. To ensure reproducible and exhaustive Y2H results, the library was screened at saturation using an optimized mating procedure that allowed for testing on average 50 million interactions per screen, corresponding to a 5-fold coverage of the library. As a consequence, multiple, independent fragments of the same interactant could be isolated, enabling the delineation of a minimal interacting domain and the computation of a confidence score.

We identified 4,089 protein-protein interactions (PPI) connecting 2,412 human proteins. 1,212 PPI (30%) are of high confidence and connect 998 proteins. Interestingly, 54 additional human oncogenes or tumor suppressors were identified in the screening, and a sub-network of 1089 PPI linking at least one cancer protein was derived from the full map. The features of the drosophila and human parallel interaction maps were compared. Orthology relationships were combined with Y2H data and the conservation of interaction networks across evolution was examined. Experimentally determined interacting domains were also systematically confronted to known functional domains in both species, pointing to a significant enrichment of domains involved in protein binding. Finally, both experimental protein interaction maps were extended by including protein interaction data from the literature. We believe that this protein interaction map centered on human cancer signaling pathways will be an invaluable resource for researchers in the field.

References :

1. Formstecher et al., 2005, Genome Research, 15:376

2. Futreal et al., 2004, Nature Review Cancer, 4(3):177

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4016.
Abstract 4028: A consolidated microplate-based workflow for monitoring intracellular calcium mobilization and mitochondrial membrane potential changes
Authors: Shen Dee , Patton Wayne
Published: February 2011

The interactions between mitochondrial function and Ca2+ homeostasis are very complex. Mitochondria are known to serve as dynamic calcium storage compartments, and also form intimate contacts with other calcium-handling organelles, such as the endoplasmic reticulum. Calcium enters mitochondria, not only during pathological processes, but also during physiological calcium responses. This organelle does not simply passively buffer excessive calcium but actively utilizes calcium signals to regulate integral cell activities, such as oxidative phosphorylation and the Kreb's cycle. A consolidated microplate-based workflow is described for assessing the relationship between mitochondria and global calcium responses. Two novel small molecule probes were implemented in the assay. Mitochondrial membrane potential was monitored using a dual-emission fluorescent probe that is roughly 10-25-fold more sensitive than the conventional carbocyanine dye, JC-1. Calcium mobilization was examined using a novel fluorescent calcium indicator dye that is significantly brighter than Fluo-4 AM dye. The assay is readily performed using a fluorescence microplate reader equipped with a dual-reagent dispenser. Proof-of-principle studies were performed by evaluating cell response to two agents, diazoxide, a putative opener of the potassium-ATP channel, and BML-258, a novel sphingosine kinase-1 inhibitor. The assay workflow is suitable for monitoring calcium mobilization and mitochondrial membrane potential changes across a broad spectrum of biological targets. The easy-to-use protocol is automation-friendly and can be performed in a convenient 96-well or 384-well microplate format. The assay should provide insight into the interaction between calcium signaling and mitochondrial energetic status, relevant to a range of phenomena, including ischemia/reperfusion injury. It should also serve as a high-throughput approach for identifying specific small molecule pharmacological agents targeted to this complex interrelationship.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4028.
Abstract 4031: Elevated expression of microRNAs in pancreatic carcinomas is correlated with poor prognosis
Authors: Greither Thomas , Grochola Philipp , Udelnow Andrej , Lautenschläger Christine , Eckert Alexander W. , Würl Peter , Taubert Helge W.
Published: February 2011

Pancreatic cancer is the eighth most common cancer and has an overall 5-year survival rate lower than 10 %. Due to their ability to regulate gene expression, microRNAs can act as oncogenes or tumour-suppressor genes and so have garnered interest as possible prognostic and therapeutic markers during the last decade. However, the prognostic value of microRNA expression in pancreatic cancer has not been thoroughly investigated.

We measured the levels of miR-155, miR-203, miR-210, miR-216, miR-217 and miR-222 by quantitative RT-PCR in a cohort of 56 microdissected pancreatic ductal adenocarcinomas (PDAC). These microRNAs were chosen as they had previously been shown to be differentially expressed in pancreatic tumours compared to normal tissues.

The possible association of microRNA expression and patients’ survival was examined using multivariate Cox's regression hazard analyses. Interestingly, significant correlations between elevated microRNA expression and overall survival were observed for miR-155 (RR = 2.50; p = 0.005), miR-203 (RR = 2.21; p = 0.017), miR-210 (RR = 2.48; p = 0.005) and miR-222 (RR = 2.05; p = 0.035). Furthermore, tumours from patients demonstrating elevated expression levels of all four microRNAs possessed a 6.2-fold increased risk of tumour-related death compared to patients whose tumours showed a lower expression of these microRNAs. This study provides the first evidence for an oncogenic activity of miR-155, miR-203, miR-210 and miR-222 in the development of pancreatic cancer as has been reported for other tumour types. Furthermore, the putative target genes for these microRNAs suggest a complex signalling network that can affect PDAC tumourigenesis and tumour progression.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4031.
Abstract 4038: Identification of microRNAs involved in colorectal cancer progression
Authors: Gariboldi Manuela , Lampis Andrea , Sokolova Viktorija , Pizzamiglio Sara , Reid James F. , Zanutto Susanna , Bertan Claudia , Perrone Federica , Verderio Paolo , Leo Ermanno , Pierotti Marco
Published: February 2011

MicroRNAs (miRNAs) are negative regulators of gene expression during various basic biological processes such as development, cellular differentiation, proliferation and apoptosis. Altered expression of miRNAs is associated with development and progression of various human cancers, where they mainly regulate the translation of oncogenes and of tumor suppressor genes. In colorectal cancer (CRC), these regulators complement the Vogelstein multi-step model of pathogenesis and have the potential of becoming a novel class of tumor biomarkers and of therapeutic targets.

To identify miRNAs deregulated in CRC that could be used for diagnosis and prognosis, we examined by Quantitative Real-Time PCR the global expression of 378 mature miRNAs in 40 CRCs and their paired non-tumor tissues. Non-parametric confidence intervals for each miRNA where calculated and twenty-four miRNAs whose expression is significantly altered in tumors respect to the normal tissue where identified (Bonferroni P<0.05). Part of them, such as miR-21, miR-31 or miR-183 has been previously reported in CRC. MiR-135b, with the highest expression in the tumors is a promising tumor related miRNA. We subsequently evaluated the association of the 24 miRNAs with different clinical characteristics of the samples such as tumor site, disease status, TNM staging, expression of CEA and CA199, and presence of alterations in the molecular markers of CRC progression (APC, TP53, KRAS and loss of 18q arm). MiRNAs differently expressed among colon and rectum or higher in late stage tumors were identified. Expression levels of miR-31 were correlated to that of CA199 and miR-18a, miR-21 and miR-31 were associated with mutations in APC gene. Gene target predictions of these miRNAs pointed mainly the pathways related to CRC progression such as Wnt/TGFb, MAPK and TP53 signaling pathways. In addition, many of these miRNAs were predicted to target genes in the Vogelstein model.

This study has identified promising miRNAs to be used as biomarkers (for example in plasma) in diagnostic and prognostic settings of CRC. And, in addition it shows their direct involvement in the regulation of driving events of CRC carcinogenesis which should be explored further in biological models.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4038.
Abstract 4041: Lipofection of the designed plasmid expressing microRNA-7 has antitumor effect in gefitinib-resistant lung adenocarcinoma in vivo
Authors: Rai Kammei , Takigawa Nagio , Ito Sachio , Kashihara Hiromi , Ichihara Eiki , Kiura Katsuyuki , Yasuda Tatsuji , Shimizu Kenji , Tanimoto Mitsune
Published: February 2011

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) is strikingly effective in tumors harboring activating EGFR mutations since the tumor cells depend on “oncogene addiction” to EGFR signaling that is inhibited effectively by a change in the affinity of ATP and EGFR-TKI to ATP binding site of mutated EGFR. Unfortunately, approximately 50% of patients who responded to EGFR-TKI acquired the resistance to EGFR-TKI due to additional mutation T790M in EGFR. Since T790M mutation restores the affinity of ATP to the ATP binding site of mutated EGFR, the oncogene addiction by EGFR pathway is presumed to sustain in the gefitinib-resistant cells harboring T790M mutation. MicroRNA-7 (miR-7) is predicted to suppress EGFR expression by targeting three sites in the 3′-untranslated region (UTR) of EGFR mRNA in silico. It was proved to be a potential tumor suppressor in EGFR-TKI-resistant lung adenocarcinoma cells by targeting the 3′-UTR of EGFR mRNA (#1396: Annual AACR meeting, 2009). The object of this experiment is to elucidate whether the miR-7 is effective or not in vivo. We established gefitinib-resistant lung adenocarcinoma cell line (RPC-9) carrying a secondary mutation T790M (Cancer Res 67: 7807-14, 2007) from gefitinib-sensitive EGFR mutated (delE746-A750 in exon 19) PC-9 cell line. RPC-9 cells (2 × 106) were injected s.c. into the backs of the female athymic mice at 7 week of age. At 1 week after injection, the mice were randomly assigned into one of three groups (five mice per group) that received either 3 μg/body of miR-7 expressing plasmid, control plasmid or mock using cationic liposome by direct injection. Marked tumor regression was observed within 2 weeks, and 60% of xenograft tumors disappeared. The immunostaining of residual tumor showed the suppression of EGFR expression on the tumor cells. To our knowledge, this is the first report to show the dramatic effect of miR-7 against EGFR-TKI resistant lung cancer model in vivo.Tumor size after treatment with miR-7 Day 1Day 5Day 11Day 18miR-710094.9445.7914.62*Control100144.0103.3242.6*tumor size(%), *p=0.00253, tumor diminished macroscopically in miR-7 group(3/5)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4041.
Abstract 4043: Targeting microRNA for the prevention and treatment of prostate cancer
Authors: Kumar Raj , Koduru Srinivas , Srinivasan Sowmyalakshmi , Rohr Jurgen , Damodaran Chendil
Published: February 2011

MicroRNAs (miRs) are small 20-24 nucleotide non-coding RNA molecules that are involved in post-transcriptional regulation of gene expression in multicellular organisms by affecting both stability and translation of mRNAs. The expression and activity of miRs vary in specific cell types and disease states. Several groups have investigated and determined the strong association between specific expression patterns of miRs and various stages of prostate cancer. Recent findings have highlighted the strong correlation between overexpression of miRs (21, 210, 125b, 221 and 222) and cancer development and progression. In this study we determined whether modulation of miR expression using a natural compound suppresses PCa growth. Previously we published that psoralidin inhibits prostate tumor growth and progression in both in vitro and in vivo models. While studying the expression patterns of a panel of miRs in control and psoralidin treated prostate cancer cells we found that psoralidin significantly inhibits expression of two specific miRNAs, miR-21 and miR-210 in turn resulting in the inhibition of EGFR-, MAPK- and PI3K-mediated prosurvival signaling and induction of JNK-mediated apoptosis in prostate cancer cells. Currently, we are overexpressing and inhibiting these two miRs to determine whether it modulates psoralidin-mediated effects on prostate cancer cells. The outcome of our study may provide encouraging data that may enable us to exploit miRs as therapeutic targets for the treatment of various malignancies.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4043.
Abstract 4045: Restoring miR-15a/16 in the NZB mouse model of chronic lymphocytic leukemia reduces disease and enhances drug sensitivity
Authors: Salerno Erica , Yuan Yao , Coffman Frederick D. , Scaglione Brian J. , Brown Brian D. , Bonci Desiree , Raveche Elizabeth S.
Published: February 2011

Alterations in the human 13q14 genomic region containing the microRNAS, mir-15a and mir-16-1 are present in a majority of human chronic lymphocytic leukemia (CLL). We have previously found the development of CLL in the New Zealand Black (NZB) murine model to be associated with a point mutation in the primary mir-15a/16-1 region, as well as a decrease in both mature miR-16 and miR-15a levels. Addition of exogenous miR-15a and miR-16 led to an accumulation of cells in G1 in both normal B cell and NZB-derived malignant B-1 cell lines. However, the NZB line had a significantly greater G1 accumulation, suggesting a restoration of cell cycle control upon exogenous miR-15a/16 addition. Our experiments demonstrated a reduction in protein levels of cyclin D1, a miR-15a/16 target and cell cycle regulator of G1/S transition, in the NZB cell line following miR-15a/16 addition. These miRNAs were shown to directly target the cyclin D1 3′UTR using a GFP lentiviral sensor. In vivo restoration of NZB with a miR-16-producing lentiviral vector resulted in the depletion of endogenous-transduced malignant aneuploid B-1 cells. In addition, experimentally increasing miR-16 in either a drug-resistant malignant NZB B-1 cell line or ex vivo splenic NZB B-1 cells augmented apoptosis induction by the following agents: genistein, a soy isoflavone tyrosine kinase inhibitor; nutlin, an MDM2 antagonist; and 2-fluoroadenine-9-B-D-arabinofuranoside, a purine nucleotide analog. Our data support a role for miR-16 in apoptosis induction, either alone, or in synergy with chemotherapeutic agents to increase their efficacy in murine CLL cells.

This research was funded in part by the NJCCR pre-doctoral fellowship awarded to E.S.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4045.
Abstract 4047: Mining of microRNA signatures in endometrial cancer
Authors: Wong Yick Fu , Cheung T.H. , Yim S.F. , Lau T.S. , Chan L.K.Y. , Ng S.W. , Chung T.K.H.
Published: February 2011

[Introduction] Endometrial cancer has now become the most common gynaecologic malignancy worldwide. About 80-90 % of endometrial cancer are endometrioid endometrial adenocarcinomas (EEC). MicroRNAs (miRNAs) are short, non-coding RNAs that regulate gene expression by inducing cleavage of the targeted messenger or by inhibiting translation. Our entire study was to mine miRNA signatures involved in EEC tumorigenesis and progression. Here we report a part of the results that is on miRNAs associated with endometrial cancer progression in vitro.

[Methods] We examined the expression of 365 mature miRNAs by real-time RT-PCR based plate assays in 52 EEC including 30 advanced- and 22 early-stage tumors, and compared that to 30 normal counterparts. To know about the deregulated miRNAs associated with endometrial cell behavior, miRNA inhibitor and pre-miRNA were used to inhibit up-regulated miRNA and induce down-regulated miRNA in highly invasive endometrial cancer cell line HEC-1A cells. Quantitative TaqMan RT-PCR was performed to measure miRNA levels in cells before and after treatment, respectively. Migration and invasion assays were carried out to evaluate effects of those deregulated miRNAs on invasion of endometrial cancer cells in vitro.

[Results] Supervised clustering analysis of global miRNA expression in endometrial caner and normal control identified 17 miRNAs up-regulated and 2 miRNAs down-regulated, respectively, in EEC. When the miRNA expression profiles in advanced stage tumors were compared to those in early stage tumors, 6 miRNAs were significantly over-expressed and one miRNA under-expressed in advanced stage tumors. Cell migration and invasion assay showed the ability of cell migration and invasion was significantly inhibited by anti-miR-7, anti-miR-194, anti-miR-449b and pre-miR-204 in HEC-1A cells in vitro.

[Conclusion] Unique miRNA signatures seem to be existed in ECC. Deregulated miRNAs miR-7, miR-194, miR-204 and miR-449b might mediate endometrial cancer progress. These miRNAs are potentially to be developed as novel diagnostic and/or prognostic molecular markers as well as candidate therapeutic targets of EEC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4047.
Abstract 4062: Human peptide deformylase expression and activity in various cancers
Authors: Reindl Katie M. , Sorensen Karly
Published: February 2011

Human peptide deformylase (hsPDF) expression has been detected in human cells, but it is unclear if the enzyme is functional. Our hypothesis is that hsPDF is functional in human cells. The objectives of this study were to 1) quantify hsPDF transcript levels in normal and cancerous cells and tissues and 2) determine the effects of hsPDF inhibition on the proliferation of normal and cancerous cells. Quantitative real-time PCR was used to measure hsPDF mRNA levels in normal and cancerous cell lines and tissues. Increases in hsPDF mRNA expression were seen in most cancer cell lines compared to the normal. hsPDF was up-regulated in most, but not all, cancer tissues compared to the normal, with the greatest differences observed in breast, colon, and lung tissues. Various cancer and normal cell lines were treated with increasing concentrations of a known PDF inhibitor, actinonin, and cell proliferation was assessed. The antiproliferative effects of actinonin were greater in prostate, colon, and breast cancer cell lines than in the normal cell lines. These results suggest that hsPDF is up-regulated in certain cancers, and that it may play an active role in the proliferation of human cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4062.
Abstract 407: Molecular characterization of invasive subpopulations from an esophageal squamous cell carcinoma cell line
Authors: Wu I-Chen , Chen Yu-Kuei , Wu Deng-Chyang , Wu Ming-Tsang
Published: February 2011

Background: Once diagnosed, esophageal cancer has a very low overall 5-year survival rate. This study investigates the mechanisms behind the invasiveness and severity of esophageal squamous cell carcinoma (ESCC). Materials and Methods: We used Transwell Invasion Chamber to subdivide one Taiwanese esophageal squamous cancer cell line CE81T/VGH into sublines (CE81T-0, CE81T-1, CE81T-2, CE81T-3, and CE81T-4) in four rounds of assays, identified the most invasive ones, and measured various factors related to their invasiveness. Results: CE81T-1, CE81T-2, CE81T-3, and CE81T-4 sublines were significantly more invasive than the parental cell (CE81T/VGH) and CE81T-0 subline. CE81T-1 and CE81T-4, the sublines we chose to study further, had significantly greater colony forming ability (3.5-2.7 fold) and wound migrating activity (1.95-2.6 fold) than the parental cell in vitro (p< 0.01). They also displayed greater tumorigenesis in immunodeficiency BALB/c Foxlnn mice than the parental cell. We found an inverse correlation between expression of TIMP-2 (tissue inhibitor of metalloproteinase-2) and invasive ability and a significant positive correlation between expressions of MMP-1 (matrix metalloproteinase-1), vimentin, and p-Src (pY416) in these cell lines and their invasiveness (all p< 0.05). Conclusion: The subline model may be used to study the molecular and genetic mechanisms underlying the invasion and metastasis of ESCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 407.