bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3046: Phosphodiesterase 4D (PDE4D) as a new drug target for prostate cancer
Authors: Hammer Kimberly D.P , Domenech Maribella , Beebe David , Marker Paul C.
Published: June 2012

Our laboratory used an insertional somatic mutagenesis screen in mice to identify novel genes that may drive prostate cancer initiation and progression. Using this screen, we have identified candidate prostate cancer genes including the candidate oncogene phosphodiesterase 4D (PDE4D). Investigation of PDE4D showed that it was over-expressed in human prostate cancer cell lines and in human prostate cancers. Stable knockdown of PDE4D by shRNA reduced the growth and migration of prostate cancer cells in vitro and in vivo. PDE4D has been implicated in the pathogenesis of chronic obstructive pulmonary disease (COPD). Several small molecule PDE4D selective inhibitors have been developed and tested in clinical trials for COPD. Although these studies showed that the inhibitors had modest efficacy as a therapy for COPD, they confirmed that the inhibitors were safe for use in humans at doses that inhibited PDE4D in vivo. Based on our previous studies with PDE4D, we postulated that PDE4D small molecule inhibitors would be candidate anti-prostate cancer drugs. Daily oral administration of PDE4D inhibitors, NVP-ABE171 or cilomilast, for 6 weeks significantly reduced the growth of LNCaP-C4 prostate cancer xenografts in immune-compromised mice. Under in vitro conditions in which prostate cancer cell growth was inhibited by either NVP-ABE171 or cilomilast, we also observed changes in the activation of potential downstream molecular targets of PDE4D inhibition. Affected pathways included the sonic hedgehog and mitogen-activated protein kinase pathways. We also tested the potential role of the PDE4D pathway in prostate cancer stromal-epithelial interactions by modeling these interactions using novel microchannel co-culture devices. In this context, PDE4D inhibitors reduced the activation of Shh pathway target genes in mesenchymal cells and reduced the proliferation of epithelial cancer cell lines, supporting the potential importance of the PDE4D pathway for prostate cancer stromal-epithelial interactions. We are continuing in vitro and in vivo studies with these candidate drugs to further support the use of NVP-ABE171 and cilomilast as anti-prostate cancer drugs and to understand their mechanism of action.

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 3046. doi:10.1158/1538-7445.AM2011-3046
Abstract 3065: Is cytology a suitable source of NSCLC tumor material for EGFR mutation testing
Authors: Dalgliesh Gillian L. , Donald Emma , Sahoo Rashmita , Patil Geeta , Augustus Meena , McCormack Rose , Ellison Gillian
Published: June 2012

Somatic mutations in the epidermal growth factor receptor (EGFR) affect non small cell lung cancer (NSCLC) sensitivity to the EGFR tyrosine kinase inhibitor gefitinib. In many countries, prospective testing of NSCLC patients for EGFR mutations is required to determine patient suitability for gefitinib. Mutation profiling is usually performed on DNA extracted from tumour biopsy samples, however the acquisition of biopsy material from NSCLC patients is often difficult and, in some cases is not available, as diagnosis is routinely made by other means. Diagnostic cytology samples, obtained by fine needle aspiration (FNA), may provide an additional source of tumour material for molecular analysis. The aims of this project were to assess the suitability of FNA for EGFR mutation testing and optimisation of DNA extraction and mutation detection methods suitable for analysis of FNA material. FNA smears of 3 human cell lines, obtained from mouse xenografts, were used to optimise DNA extraction methods. DNA was extracted from diagnostic FNA smears or paraffin embedded FNA derived cells, from 25 NSCLC patients of Indian origin, using the optimised protocols. Following successful DNA extraction from all patient samples, EGFR mutation status was determined by both DNA sequencing and using the therascreen EGFR Mutation Test Kit (L858R and exon 19 deletion mutations only). Twelve of the 25 patients harboured an EGFR mutation. For this sample type, the therascreen EGFR Mutation Test Kit was more effective at identifying mutation positive patients with only 4 of 12 detected mutations also evident by DNA sequencing. DNA extracted using a spin column based kit and quantified by real time PCR prior to performing molecular analysis gave the most robust, reproducible results. In conclusion, these data suggest that diagnostic FNA smears or cell blocks are suitable for assessing EGFR mutation status of NSCLC patients and may prove a useful source of tumour material when biopsies are unavailable.

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 3065. doi:10.1158/1538-7445.AM2011-3065
Abstract 3074: Identification of a predominant pattern of co-regulation among kinetochore genes; prospective regulatory elements, and association with genomic instability
Authors: Reinhold William C. , Erliandri Indri , Liu Hongfang , Zoppoli Gabriele , Pommier Yves , Larionov Vladimir
Published: June 2012

The NCI-60 cell line panel is the most extensively characterized set of cells in existence. It provides multiple forms of data on different cancer types, and has been used extensively as a screening tool for drug discovery. Previously, the potential of this panel has not been applied to the fundamental cellular processes of chromosome segregation. In the current study, we used data from multiple microarray platforms accumulated for the NCI-60 to characterize an expression pattern of genes involved in kinetochore assembly. This analysis revealed that 17 genes encoding the constitutive centromere associated network of the kinetochore core (the CCAN complex) plus four additional genes with established importance in kinetochore maintenance (CENPE, CENPF, INCENP, and MIS12) exhibit similar patterns of expression in the NCI-60, suggesting a mechanism for co-regulated transcription of these genes which is maintained despite the multiple genetic and epigenetic rearrangements accumulated in these cells (such as variations in DNA copy number and karyotypic complexity). Several potential regulatory influences are identified for these genes, including transcription factors and microRNAs. Thus, our results provide a template for experimental studies on regulation of genes encoding kinetochore proteins, the process that, when aberrant, leads to the aneuploidy that is a hallmark of many cancers. We propose that the comparison of expression profiles in the NCI-60 cell line panel could be a tool for the identification of other genes groups whose products are involved in the assembly of organelle protein complexes.

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 3074. doi:10.1158/1538-7445.AM2011-3074
Abstract 3075: A novel transcriptional role of spindle assembly checkpoint protein Cdc20 regulating the expression of mitotic ubiquitin carrier protein UbcH10
Authors: Nath Somsubhra , Banerjee Taraswi , Sen Debrup , Das Tania , Roychoudhury Susanta
Published: June 2012

Progression through mitosis requires precisely timed ubiquitin-dependent degradation of specific substrates. UbcH10, an ubiquitin-conjugating enzyme, plays a crucial role with anaphase promoting complex/ cyclosome (APC/C) in progression of and exit from mitotis. Similarly, spindle assembly checkpoint (SAC) protein Cdc20 plays an important role in transition from metaphase to anaphase by activating APC/C. Cdc20 itself gets ubiquitinated by UbcH10 and thus activated to bind with APC/C. Any defect in chromosome alignment activates SAC by blocking Cdc20 and thus anaphase transition is arrested. Earlier our lab reported that Cdc20 overexpression led to defective SAC resulting in aneuploidy. This prompted us to examine the regulation of UbcH10 expression in Cdc20 upregulated condition.

Initially, we observed a correlation in the expression of both Cdc20 and UbcH10 in different primary tumors and cancer cell lines. Next, a dose-dependent increase in endogenous UbcH10 levels was found upon ectopic expression of Cdc20. Alternatively, CDC20 knockdown by siRNA led to downregulation of endogenous UbcH10. Activation of UBCH10 promoter-luciferase activity by Cdc20 indicated the regulation at transcriptional level. ChIP assay showed the presence of Cdc20 on UBCH10 promoter leading to chromatin remodelling by interacting with CBP/p300. APC/C-CBP/p300 complex is reported to be involved in transcription regulation. Here, we established the presence of APC/C on the UBCH10 promoter using ChIP assay. Further from ChIP and Co-IP assay we established that Cdc20 regulates APC/C and CBP/p300 association and thus modulates APC/C-CBP/p300 mediated transcriptional regulation of UBCH10. Also, we found cell cycle specific regulation of expression of UbcH10 by Cdc20. Finally, Cyclin B1 degradation assay showed physiological relevance of Cdc20 mediated UbcH10 regulation on mitotic progression.

In summary, our data showed a novel role of Cdc20 as a transcription regulator and it regulates UbcH10 expression in a cell cycle specific manner. The co-regulated expression of both these proteins maintains proper mitotic progression which otherwise may results in aneuploidy.

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 3075. doi:10.1158/1538-7445.AM2011-3075
Abstract 3087: Regulation of p53 stability and activity by ribosomal protein S27a
Authors: Sun Xiao-Xin , DeVine Tiffany , Dai Mushui
Published: June 2012

Ribosomal proteins (RPs) play a critical role in tightly coordinating p53-dependent cell cycle checkpoint with ribosomal biogenesis. A number of RPs, including L5, L11, L23, L26, and S7, specifically bind to MDM2 and inhibit MDM2-mediated p53 ubiquitination and degradation, leading to p53 activation, in response to nucleolar stress triggered by the perturbation of ribosomal biogenesis. Here, we report S27a, a ribosomal protein from small subunit of ribosome and synthesized as an ubiquitin C-terminal extension protein (CEP80), as a novel MDM2 regulator. S27a interacts with MDM2 at the central acidic domain of MDM2 and suppresses MDM2-mediated p53 ubiquitination, leading to p53 activation and cell cycle arrest. Knockdown of endogenous S27a significantly attenuated p53 activation by treatment with a low dose (5 nM) of actinomycin D (Act D), which specifically inhibits RNA polymerase I activity leading to inhibition of rRNA synthesis and perturbation of ribosomal biogenesis, suggesting that S27a is a non-redundant MDM2 regulator in response to nucleolar stress. Interestingly, we also show that MDM2 ubiquitinates S27a, leading to protesomal degradation of S27a, in response to Act D treatment, suggesting that MDM2, in turn, controls S27a levels in response to nucleolar stress. Taken together, our results reveal that S27a regulates p53 activity in response to nucleolar stress via a novel and dynamic interplay between S27a and MDM2.

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 3087. doi:10.1158/1538-7445.AM2011-3087
Abstract 3121: Study of leukemia inhibitory factor (LIF) system in a murine mammary tumor comprising luminal and myoepithelial cells
Authors: Krasnapolski Martin A. , Quaglino Ana , Kordon Edith , Bal de Kier Joffe Elisa D.
Published: June 2012

The presence of LIF, a cytokine that activates JAK/STAT and ERK pathways, its receptor (LIFRb) and the constitutive activation of Stat3 (a molecule downstream LIFRb) have been described in tumors, primary cultures and several cancer-derived cell lines, suggesting a role for this pathway in tumor progression. Also, Stat3 activation was widely related to an immunosuppressive phenotype in many tumor models.

From the murine mammary adenocarcinoma M38 comprising luminal (LEP) and myoepithelial (MEP) cells (both transformed) we derived LM38-LP (LEP & MEP), LM38-HP (epithelioid) and LM38-D2 (MEP) cell lines. While the mixed cell line presents a malignant behavior, the last two show a lower growth rate and metastatic potential than the parental tumor. Previously we demonstrated that this lower malignancy is partially caused by their inability to suppress the host's immune system, underscoring the importance of cellular interactions in complex tumor behaviors.

Our purpose was to evaluate LIF and LIFRb mRNA expression levels and to study LIF effects on Stat3 and ERK activation, as well as on the proliferation and clonogenic capacity in the LM38 family of cell lines.

Using RT-PCR we could see that LM38-LP cell line expresses high LIF and LIFRb mRNA levels, LM38-HP expresses LIF mRNA but not it receptor's mRNA and LM38-D2 has low levels of both molecules. By Western blot we studied LIF (80 ng/ml) effect over Stat3 and ERK activation. While in LM38-LP and LM38-D2 cells Stat3 and ERK phosphorylation was rapidly induced at 5 min, in LM38-HP cells, that don't express LIFRb, no response was detected. Then, we evaluated the effect of a prolonged 48 h treatment with LIF (1, 5, 20 and 80 ng/ml) on cell viability using MTS assay. While LIF slightly reduced cell viability (20%) in LM38-LP and LM38-D2 cell lines, LM38-HP cells were not affected. Finally, since LIF is also involved in stem cell maintenance, the effect of LIF in a low density/clonogenic setting was evaluated. While in LM38-LP and LM38-D2 cell lines LIF significatively (p<0.05) increased the number of colonies, LM38-HP cells were unaffected.

We can conclude that LIF could be involved in LEP-MEP interaction and although it induced a subtle viability reduction, LIF could be nevertheless promoting tumor progression by increasing the number of clonogenic cells. In addition, LIF-induced Stat3 activation could be the responsible of the immunosuppresive characteristics of the mix tumor cell line, a proposal that deserves future studies.

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 3121. doi:10.1158/1538-7445.AM2011-3121
Abstract 3123: Differential effects of polyoma virus middle tumor antigen mutants upon gap junctional, intercellular communication
Authors: Greer Samantha N. , Geletu Mulu , Raptis Leda H.
Published: June 2012

Gap junctions are channels that connect the cytoplasm of adjacent cells. Gap junctional, intercellular communication (GJIC) is blocked in cells transformed by oncogenes such as the middle Tumor antigen of polyoma virus (mT), an oncoprotein which associates with and is tyrosine-phosphorylated by cSrc family members. Specific phosphotyrosines provide docking sites for the phosphotyrosine binding domain of Shc (mT-tyr250) and the SH2 domain of the phosphatidylinositol 3-kinase (mT-tyr315), resulting in the activation of their downstream signaling cascades, Ras/Raf/Erk and PI-3 kinase/Akt, respectively.

To examine the effect of these mT-initiated pathways upon gap junctional communication and neoplasia, GJIC was assessed in mT-mutant-expressing, rat liver epithelial T51B cells which normally have extensive GJIC, using a novel technique of in situ electroporation we developed. The results show that although low levels of wt-mT are sufficient to interrupt gap junctional communication, GJIC suppression still requires an intact tyr250 site, that is activation of the Ras pathway. In sharp contrast, activation of the PI-3 kinase pathway is not required for GJIC suppression. It is remarkable that T51B cells expressing a mutant deficient in binding of Shc, hence activation of the Ras pathway, are still morphologically transformed and do grow into tumors in syngeneic rats, albeit with an altered morphology. These results indicate that GJIC suppression requires Ras/Erk but not PI-3 kinase/Akt signalling, and is independent of neoplastic conversion in rat liver epithelial 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 3123. doi:10.1158/1538-7445.AM2011-3123
Abstract 3161: Increased expression of MUC1 and claudin-10 is associated with poor prognosis of patients with gastric cancer
Authors: Park Keon Uk , Kim Jin Young , Do Young Rok , Song Hong Suk , Hwang Ilseon , Ha Eunyoung , Ryoo Hun Mo , Min Young Joo
Published: June 2012

Background: Mucins are high molecular weight glycoproteins that play important roles in carcinogenesis and tumor invasion. Several studies have shown the aberrant expression profile of mucins in gastric cancer. Claudins are the proteins that participate in the formation of tight junctions. Tight junction proteins are believed to be involved in the regulation of proliferation, differentiation, and other cellular functions. Expression of claudin family in gastric cancers has been studied by many laboratories, but the results are conflicting. Therefore, we investigated the potential of MUC and claudin family to be used as a prognostic marker in gastric cancer.

Methods: One-hundred eighty gastric adenocarcinoma patients who underwent surgical resection and had not received any pre- or -post surgery therapy, were selected for this study. Among the 180 gastric cancer samples tested here, 63 (34%) were early gastric cancer, and 117 (66%) were advanced. We made tissue microarrays with paraffin-embeded formalin fixed blocks of gastric cancer and these microarrays were evaluated for phenotypic expression of MUC1, MUC4, Claudin 1, 3, 4, 5, 7 and 10 using anti-human rabbit or mouse polyclonal antibody. The expression levels were correlated with key clinicopathologic features and patient outcomes.

Results: There was no significant difference of MUC and claudin expression between early and advanced gastric cancer. Gastric cancer patients with increased MUC1 expression were significantly associated with shorter overall survival as compared to those with low expression of MUC1 (log rank, p=0.039). MUC4 expression were significantly associated with decreased overall survival only in N0 patients (log rank, p=0.046). Increased expression of Claudin-10 was significantly associated with shorter disease free survival (log rank, p=0.038). Claudin 1,3,4,5 and 7 expression was not associated with significant difference of overall survival or disease free survival.Conclusion: Our present findings show that the increased expression of MUC1 and claudin-10 is a prognostic indicator of poor survival of patients with early and advanced gastric cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3161. doi:10.1158/1538-7445.AM2011-3161
Abstract 3184: Diagnostic value of Annexin A1 as a marker for myoepithelial cells in normal breast glands but is lost in malignant breast glands
Authors: Elshimali Yahya I. , Wu Yanyuan , Chen Daniel , Vadgama Jaydutt V.
Published: June 2012

Introduction: Annexin AI belongs to the annexin family of Ca2+-dependent phospholipid-binding proteins. Annexin I has been of interest for use as a potential target for anticancer drugs. Upon induction by modified NSAIDS and other potent anti-inflammatory drugs, annexin I inhibits the NF-κB signal transduction pathway, which is exploited by cancerous cells to proliferate and avoid apoptosis. ANXA1 inhibits the activation of NF-κB by binding to the p65 subunit.

Methods: We used immunohistochemistry stain of Annexin A1 monoclonal antibody to study its expression in 100 cases of breast tissue, 50 cases of normal tissue vs. 50 cases of malignant tissue, using tissue micro array (TMA).

Results: Our analysis show a decreased expression of Annexin A1 in malignant tissues compared to non-malignant tissues. Annexin A1 stains myoepithelial cells in 95 % of normal glands, but it is absent in 90 % of the periphery malignant glands. This is most likely due to the absence of the myoepithelial cells.

Conclusion: We conclude that Annexin A1 has diagnostic value in breast pathology by staining myoepithelial cells of normal gland in addition to its characteristic in staining of normal ductal cells at different intensity. Annexin A1 could be used as a biomarker for myoepithelial cells in surgical pathology practice.

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 3184. doi:10.1158/1538-7445.AM2011-3184
Abstract 3188: Discovery and validation of non-invasive biomarkers for benign prostate hyperplasia and prostate cancer
Authors: Curatolo Adam S. , Roy Roopali , Jedinak Andrej , Loughlin Kevin , Meyers Michael , Libermann Towia A. , Dillon Simon , Bhasin Manoj , Moses Marsha A.
Published: June 2012

Although the PSA test is successfully used to determine how well a prostate cancer patient is responding to therapy, it cannot reliably distinguish between the presence of prostate cancer and benign prostate diseases such as benign prostatic hyperplasia (BPH) or prostatitis. These conditions and early prostate cancer can all result in low to moderate levels of PSA in the blood. The absence of sensitive and specific biomarkers that can distinguish between benign conditions and prostate cancer often leaves patients and their doctors with little recourse than to utilize invasive approaches to diagnose prostate cancer that can result in harmful and undesirable side effects. With the goal of identifying and validating non-invasive biomarkers that can distinguish between neoplastic and non-neoplastic disease of the prostate, we have utilized iTRAQ (isobaric tag for relative and absolute quantitation) as our technique of choice to sensitively and accurately identify and quantify proteins that are differentially present in the urine of men with BPH and prostate cancer. This approach resulted in the identification of 26 proteins that were differentially expressed at a significant level, with high confidence, in urines obtained from men from each of the two groups of interest. These proteins represent a number of functional categories including cell assembly and organization, cell signaling, cell morphology, carbohydrate metabolism, cellular growth and proliferation, lipid metabolism, androgen and estrogen metabolism and DNA replication, recombination and repair, among others. Validation of ten proteins with the greatest differential expression between the two groups has been completed. Immunoblot analyses have confirmed the significant differential presence of these proteins in a larger number of patient samples analyzed thus far. Large scale validation studies are ongoing. These data suggest that the ability to non-invasively monitor the levels of these proteins in the urine of patients with uninformative PSA levels has the potential to provide clinically relevant information for reliable diagnosis of BPH and prostate cancer. These proteins may also serve as potential therapeutic targets for the treatment of men with BPH and prostate cancer. (Supported by The Ellison Foundation)

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 3188. doi:10.1158/1538-7445.AM2011-3188
Abstract 3202: Decreased Dkk3 protein expression is associated with a poor prognosis of cervical cancer
Authors: Lee Eun-Ju , Kim Mi Kyung , Park Hyung Moo , Lee Sang-Hoon , Kim Dong-Ho , Lee Moo-Yul , Myung Soon-Chul , Kim Won Yong
Published: June 2012

Dkk3 has tumor suppressor properties in various cancers. Recently, we reported that the mRNA level of Dkk3 gene was down-regulated by epigenetic alteration in cervical cancer and that forced expression of Dkk3 resulted in the growth inhibition of cervical cancer cells. The aim of this study was to assess the expression of Dkk3 protein in cervical cancer tissue and determine potential clinical correlations. Immunohistochemical analysis was performed on 94 cervical cancer tissues. Clinical and pathological parameters were obtained from medical record. Survival data were estimated using Kaplan-Meier estimates and compared with log-rank test where indicated. Out of 94 tissue samples, negative, weak, moderate and high expression of Dkk3 were observed in 29 (30.9%), 31 (33.0%), 18 (19.1%) and 16 (17.0%) tissue samples. Dkk3 expression was negatively correlated with FIGO clinical stage (p<0.03, Mann-Whitney test). The 5-year disease free survival of patients with a negative or weak expression was significantly lower than that of those with moderate or high expression (70.6% versus 86.3%, respectively, p=0.04, log-rank test). The result of this study showed that down-regulation of Dkk3 is associated with advanced FIGO clinical stage and poor survival outcome of cervical cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3202. doi:10.1158/1538-7445.AM2011-3202
Abstract 3225: Influence of localized vibration loading on bone targeting of bisphosphonate
Authors: Papineni Rao V. , Pan Jingyi , Geldhof Benjamin
Published: June 2012

Targeting of bisphosphanate analog to specific skeletal sites is investigated by using localized vibration loading. Breast, lung, and prostate cancer cells metastasize frequently at bone resulting in debilitating pain, which is one of the major complications in majority of patients. The palliative bone pain management includes the bisphosphanate therapy. Bisphosphonates apart from being a potent inhibitor of bone resorption, also provides pain relief and improves bone health. However these medications result in complications such as bisphosphonate-related osteonecrosis of the jaw (BRONJ). Our continued efforts in investigating site-selective skeletal targeting of bisphosphonates are primarily to establish modes to overcome such drug-related side-effects. Non-invasive near-infrared (NIR) fluorescence imaging was used to monitor binding of NIR dye conjugated bisphosphonate (osteosense) to specific skeletal sites in response to localized vibration loading. Vibration loading, was performed on multiple sets of mice- subjecting them to vibration frequency of 70 Hz for five min (5 times/week) duration at the right thigh region. Osteosense and was injected intravenously after 21-day exercise regime. Both noninvasive and ex vivo imaging show significantly increased NIR signals at the right surface of the lumbar vertebrae. The unequivocal outcome in both the cohort studies, following the localized vibration loading confirms the role of direct high-frequency vibration stimulation in bisphosphonate targeting. Histochemical analysis of the bone and the muscle describe the mechanistic role in the targeted binding of the bisphosphonate.

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 3225. doi:10.1158/1538-7445.AM2011-3225
Abstract 3281: Functional characterization of a fusion angioinhibitor molecule “Combostatin”
Authors: Boosani Chandra Shekhar , Gunda Venugopal , Laknaur Archana , Akulapalli Sudhakar
Published: June 2012

Angiogenesis, a process of new blood vessel formation is observed during pathological and physiological conditions, can be controlled by circulating endogenous angioinhibitory molecules derived from non-collagenous domains of type IV and type XVIII collagen. In the present study we developed a fusion protein (Combostatin) from functional domains of three known endogenous angioinhibitors [α1(IV)NC1 (arresten), α3(IV)NC1 (tumstatin) and type XVIII collagen (endostatin)] using recombinant DNA technology and expressed using Baculovirus expression system. We then tested the angioinhibitory functions of Combostatin in-vitro using cellular proliferation, invasion, migration and tube formation assays. Combostatin was found to inhibit endothelial cell proliferation and invasion in a dose dependent manner, similar to its parent molecules. Interestingly, inhibition of endothelial migration and tube formation by Combostatin was significantly higher compared to its parent molecules. In addition Combostatin binds to different cell surface integrins and inhibits multiple signaling pathways. Taken together, our studies identify that Combostatin is a more potent angioinhibitor molecule compared to its parent molecules. Our results support earlier studies that showed multiple angioinhibitors which are promising candidates with clinical importance compared to single inhibitor molecules. Further pre-clinical validation of Combostatin is essential to characterize its use in the treatment of vascular diseases including cancer, diabetic retinopathy etc.

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 3281. doi:10.1158/1538-7445.AM2011-3281
Abstract 331: C-MYC gene amplification in endometrial cancer is associated with aggressive tumors
Authors: Birkeland Even , Krohn Antje , Trovik Jone , Holst Frederik , Werner Henrica M.J. , Krakstad Camilla K. , Wik Elisabeth , Akslen Lars , Simon Ronald , Salvesen Helga B.
Published: June 2012

Endometrial cancer is among the most common gynaecological malignancies in industrialized countries. Although 75% of cancers are treated at an early stage, 15% to 20% recur. Due to these recurrences the demand for systemic therapies and better prognostic markers are vital. Our recent study of global genetic characterisation has identified that C-MYC amplifications associate with aggressive phenotype in uterine cancers (Salvesen et al., PNAS 2009).

The present study was conducted to validate the phenotype associated with C-MYC amplification in a large population based prospectively collected sample series from primary tumours and corresponding metastatic lesions from patients with endometrial cancer. Primary tumours from 399 patients and 80 corresponding metastatic lesions were analysed for amplification of the C-MYC gene using fluorescence in situ hybridization (FISH).

We found that 5% of primary endometrial cancer tumors have C-MYC amplifications or gains. C-MYC amplification is correlated with high FIGO stage (P = 0.003), non-endometroid subtype (P = 0.002), and high grade (P <0.001). Patients with C-MYC amplifications had a significantly poorer recurrence free- (P = 0.002) and disease specific survival (P = <0.001). These data suggest that C-MYC amplification might be one important mechanism involved in endometrial cancer tumourigenesis. Further characterization of C-MYC amplifications in corresponding metastatic lesions are in progress.

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 331. doi:10.1158/1538-7445.AM2011-331
Abstract 3356: Loss of breast epithelial hCLCA2 promotes EMT and increases risk of metastasis
Authors: Elble Randolph C. , Walia Vijay , Yu Yang , Cao Deshou , Sun Miao , Premkumar Louis
Published: June 2012

Transition between epithelial and mesenchymal states is a feature of both normal development and tumor progression. We report that expression of chloride channel accessory protein hCLCA2 is a characteristic of epithelial differentiation in the immortalized MCF10A and HMLE models, while induction of EMT by cell dilution, TGFbeta, or mesenchymal transcription factor Snail sharply reduces hCLCA2 levels. Attenuation of hCLCA2 expression by lentiviral shRNA caused cell overgrowth and focus formation, enhanced migration and invasion, and increased mammosphere formation in methylcellulose. These changes were accompanied by downregulation of E-cadherin and upregulation of mesenchymal markers such as vimentin and fibronectin. Moreover, hCLCA2 expression is greatly downregulated in breast cancer cells with a mesenchymal or claudin-low profile. These observations suggest that loss of hCLCA2 may promote metastasis. We find that higher-than-median expression of hCLCA2 is associated with a one-third lower rate of metastasis over an 18 year period among breast cancer patients compared to lower-than-median (n=344, unfiltered for subtype). Thus, hCLCA2 is required for epithelial differentiation, and its loss during tumor progression contributes to metastasis. Overexpression of hCLCA2 has been reported to inhibit cell proliferation and is accompanied by increases in chloride current at the plasma membrane and reduced intracellular pH (pHi). We found that knockdown cells have sharply reduced chloride current and higher pHi, both characteristics of tumor cells. These results suggest a mechanism for the effects on differentiation. Loss of hCLCA2 may allow escape from pHi homeostatic mechanisms, permitting the higher intracellular and lower extracellular pH that are characteristic of aggressive tumor 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 3356. doi:10.1158/1538-7445.AM2011-3356
Abstract 3357: Partial mesenchymal to epithelial reverting transition in breast and prostate cancer metastases
Authors: Chao Yvonne , Wu Qian , Acquafondata Marie , Dhir Rajiv , Wells Alan
Published: June 2012

Epithelial to mesenchymal transition (EMT) is an oft-studied mechanism for the initiation of metastasis. We have recently shown that once cancer cells disseminate to a secondary organ, a mesenchymal to epithelial reverting transition (MErT) can occur, which we postulate is to enable metastatic colonization. Despite a wealth of in vitro and in vivo studies, evidence of MErT in human specimens is rare and difficult to document because clinically detectable metastases are typically past the micrometastatic stage at which this transition is most likely evident. We obtained specimens of primary and metastatic tumors from breast and prostate cancer patients and evaluated expression of various epithelial and mesenchymal markers by immunohistochemistry. Breast cancer metastases exhibited increased expression of membranous E-cadherin and β-catenin compared to primary tumors, consistent with EMT at the primary site and MErT at the metastatic site. Expression of epithelial markers connexins 26 and 43 was also increased, particularly in brain metastases. Despite the upregulation of epithelial markers in metastases, expression of mesenchymal markers vimentin and FSP1 was mostly unchanged or only slightly decreased. Furthermore, the switch in expression of E-cadherin to N-cadherin that is sometimes detected in EMT, was not found by our immunostaining. N-cadherin detection did not reveal a consistent pattern of expression in primary or metastastic tumors. We observed similar results in prostate cancer metastases; not only was E-cadherin expression increased but also expression inversely correlated with size of the metastasis, suggesting that a second EMT may occur in the ectopic site for tumor growth or to seed further metastases. In summary, we have observed an increased expression of epithelial markers and persistence of mesenchymal markers consistent with a partial MErT that may enable a second EMT at the metastatic site. Our results suggest that cancer cells continue to display phenotypic plasticity beyond the EMT that initiates metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3357. doi:10.1158/1538-7445.AM2011-3357
Abstract 3381: AhR shRNA effectively suppresses B(a)P-mediated induction of slug modulating tumor invasion and cell migration in non small cell lung cancer cell line
Authors: Ahn Kwang-Sung , Park Juwon , Kim Kyungsun , Chae Yang Seok , Won Nam Hee
Published: June 2012

Exposure of B(a)P on lung tissue have been implicated in cancer development and its progression. Even though increasing evidences have been reported adversely effects of B(a)P in tumor progression, mechanism by which exposure of Bap on lung cancer cells increases the metastatic potentials still remains unknown. Here, we show that B(a)P triggers a marked morphological changes of Non Small Cell Lung Cancer (NSCLC) cells. These B(a)P-mediated effects are mimicked by decreasing E-cadherin resulting in loosening cell-cell contacts and by increasing Slug expression resulting in inducing epithelial mesenchymal transition (EMT). In case that NCSCLC cell was prolonged exposed to B(a)P, B(a)P-exposed cells showed increasing cell motility and invasion. They correlate with B(a)P-mediated phosphorylations of JNK and p38. And they are reverted by treatment with their inhibitors. In transient transfection analysis, H1299-snail2, constitutively expressing snail2, increased the cell motility and invasion. AhR shRNA effectively suppresses either B(a)P-mediated tumor invasion and induction of snail2. And suppression of E-caherin was found in H1299-snail2. Results of immunohistochemical stain revealed that Slug strongly expressed in invasive foci of Non Small Cell Lung Cancer tissue. Our results suggested that NSCLC cells prolonged exposed to B(a)P increased cell motility and invasiveness via regulation of expression of Sanil2 by exposure of B(a)P on NSCLC 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 3381. doi:10.1158/1538-7445.AM2011-3381
Abstract 3411: Analysis of gene profile and identification of molecular targets in endometrial cancer stem cells
Authors: Nakamura Mitsuhiro , Kyo Satoru , Zhang Xivzhi , Takakura Masahiro , Maida Yoshiko , Mizumoto Yasunari , Mori Noriko , Bono Yukiko , Sasagawa Toshiyuki , Inoue Masaki
Published: June 2012

Background; Cancer stem cells (CSCs) have been known to be the major source of tumor propagation and might be an attractive therapeutic target. Our previous study showed that CD133+ Ishikawa and MFE 280 endometrial cancer cells have the ability of tumorigenicity, self-renewal and differentiation capacity in vitro and in vivo, while CD133- cells did not. Furthermore, CD133+ cells showed more resistance to cisplatin- or paclitaxel-induced cytotoxicity than CD133- cells. These results indicate that CD133 is the CSC maker in endometrial cancer cells. In the present study, we sought to analyze gene profile and identify molecular targets in endometrial cancer stem cells. Methods; We examined microarray analysis to investigate the molecules essential for the CSCs characteristics by 3D-Gene chip (TORAY). Results; Microarray analysis of total 25,000 genes showed that about 500 genes were highly expressed in CD133+ cells, compared to CD133- cells. The ALDH1 gene was up-regulated in CD133+ cells, in accordance with the previous reported expression of ALDH1 on breast and colorectal CSCs. We also found out the elevated expression of ABC transporter genes, ABCG2, ABCG3, ABCG4, ABCG8, ABCC8 and ABCC9. It is well known that cells expressing those genes are resistant to chemotherapeutic agents and associated with side population (SP). Fluorescence activated cell sorter analysis demonstrated that the ratio of SP fraction in CD133+ Ishikawa cells is slightly higher than in CD133- Ishikawa cells (0.8% vs 0.6%). Isolated SP/CD133+ cells ells showed more extensive proliferation and tumorigenicity, compared with non-SP/CD133+, SP/CD133- and non-SP/CD133- cells. Finally, we focused on high expression of MMP14 (MT1-MMP) gene in CD133+ cells. Invasion assay revealed that CD133+ Ishikawa and MFE 280 cells have more invasive property than CD133- Ishikawa and MFE 280 cells. The inhibition of MT1-MMP gene by siRNA significantly reduced the invasive ability of CD133+ cells. These results suggest that MT1-MMP is one of the critical molecules that generate biological features of CSC and may be a potential therapeutic target in endometrial cancer. Conclusions; We demonstrated that SP/CD133+ cells might be more defined CSCs in endometrial cancer and identified important molecules essential for characterizing endometrial CSCs in which MT1-MMP plays critical roles in generating biological features of CSCs in this tumor type. These findings shed light on establishment of molecular target therapy in endometrial cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3411. doi:10.1158/1538-7445.AM2011-3411
Abstract 3427: A core invasiveness gene signature reveals reduced EMT in samples from patients with IBC, potentially regulated by decreased EGFR- and TGFb-signaling
Authors: Marsan Melike , Limame Ridha , François Katrien , Bertucci François , Ueno Nato , Lardon Filip , Pauwels Patrick , Van Dam Peter , Vermeulen Peter B. , Dirix Luc Y. , Van Laere Steven J.
Published: June 2012

Introduction. We identified a core invasiveness gene (CIG) signature that predicts the invasive properties of breast cancer cell lines. In the current study, we investigate the translational utility of the current gene signature in human breast cancer, including inflammatory breast cancer (IBC). The latter is a highly invasive and metastatic breast cancer subtype.

Materials and methods. Six publicly available gene expression data sets and a data set on 137 IBC and 252 non-IBC samples were analyzed. Each sample was classified according to sets of stromal (N=2), prognostic (N=2), stem cell (N=3), epithelial-to-mesenchymal transition (EMT) (N=3) and pathway (N=20) gene signatures. In addition, the cell-of-origin subtype classifier and our own CIG-signature were applied. Associations between the CIG-signature and clinicopathological or molecular features were searched for using univariate and multivariate analysis.

Results. We identified associations of the CIG signature (FDR<0.01) with a mammosphere-derived classifier (median Rs=0.49) and with two EMT-signatures (Core-EMT: median Rs=0.52; EMT/stromal classifier: median Rs=0.59). In addition EGFR, STAT3 and TGFb were found to be hyperactivated in CIG-positive tumors (median Rs=0.45; 0.33 and 0.60 respectively). No relations between CIG-expression and stromal gene expression, molecular subtypes or any of the clinicopathological variables were observed. Compared to non-IBC, decreased CIG-expression, TGFb- and EGFR-activation were observed in IBC (respectively P<0.0001, P<0.0001 and P=0.036). As for other EMT-associated gene signatures, both positive (Claudin-low and LYN-signatures: P=0.004 and P=0.015) and negative (EMT/stromal classifier: P=0.015) associations with IBC have been observed. When testing the core-EMT signature, no difference was found (P=0.166).

Discussion. We show that the CIG signature in breast cancer is correlated with EMT, stem cell biology and EGFR-, TGFb- and STAT3-activation. Samples from patients with IBC demonstrate ambiguous EMT-patterns, suggest that tumor cells from patients with IBC are in a specific state of cell plasticity and EMT as such is not the main mode of invasion in IBC. On the contrary, lowered TGFb signaling in IBC suggest a form of collective invasion.

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 3427. doi:10.1158/1538-7445.AM2011-3427
Abstract 3444: Enforced expression of MycN and C-Myc induces different medulloblastoma subtypes
Authors: Kawauchi Daisuke , Robinson Giles , Uziel Tamar , Rehg Jerold , Zindy Frederique , Qu Chunxu , Gajjar Amar , Gilbertson Richard J. , Roussel Martine F.
Published: June 2012

Medulloblastoma (MB), a tumor of the cerebellum, is the most common malignant brain tumor of childhood. MBs are classified histologically and molecularly into four distinct major subgroups, two of which correspond to the deregulation of major developmental pathways, Sonic Hedgehog (SHH) and WNT. Among the two other subgroups, one correlates with high levels of C-MYC expression. Using orthotopic transplantation in the cerebellum or cortex of naïve recipient mice, we showed that enforced expression of MycN in cerebellar granule neuron progenitors (CGNPs) isolated from the cerebellum of postnatal (P) day 6-7 Cdkn2c-/-, Trp53-/-, Atoh1-GFP mice induced MBs with a SHH “signature”. Interestingly, overexpression of C-Myc in the same CGNPs induced MBs with Large Cell Anaplastic (LCA) features and mRNA and microRNA profiles distinct from the SHH subgroup. In vitro analysis revealed that tumor cells were resistant to Shh antagonists, such as cyclopamine and BMP4. Ongoing experiments are focusing on understanding the differences between MycN and C-Myc function in CGNPs.

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 3444. doi:10.1158/1538-7445.AM2011-3444