bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 96: Conformational and thermodynamics analyses of the regulation of trifluoperazine on camodulin binding to Fas: Implications for cancer chemotherapy
Authors: Pan Di , Yan Qi , Chen Yabing , McDonald Jay M. , Song Yuhua 
Published: February 2011

Abstract
Fas-mediated signaling pathway is an important mechanism for apoptosis in a variety of cells, including cholangiocarcinoma and other cancer cells. The formation of death inducing signaling complex (DISC) is a critical step for Fas-mediated signaling. Recent experimental studies showed that calmodulin (CaM) binds to Fas and regulates Fas-mediated DISC formation and the binding of CaM to Fas is inhibited by the CaM antagonist, trifluoperazine (TFP) (J Cell Biochem, 103:788, 2008). The present study sought to determine the structural basis and molecular mechanisms of TFP regulation of Fas-mediated DISC formation. We used a combination of molecular dynamics simulations and binding free energy analyses to investigate the effect of TFP on CaM-Fas binding. We studied the interactions of different number(s) of TFP(s) with CaM and the effect of the resulting conformational changes of CaM on CaM/Fas binding. We found that the binding affinity of one TFP with CaM was affected by the sequential binding of other TFP(s) to CaM, which was consistent with near-UC circular dichroism data (Biochemistry 37:15300, 1998). The number of TFP(s) bound to CaM was shown to affect the conformational and motion changes of CaM, and thus directly affect CaM binding to Fas. Consistent with previous experimental observations that CaM-Fas binding was affected by TFP concentrations (J Biol Chem, 280:29964, 2005), we demonstrated that one TFP bound to CaM inhibited CaM-Fas binding while the subsequent two TFPs or four TFPs bound to CaM resulted in the recovery of CaM-Fas binding (Table 1). Results also showed that conformational and motional changes of Fas by binding to CaM were directly related to the number of TFPs bound to CaM, which could further affect Fas recruitment of FADD to the DISC. This study provides structural and thermodynamics evidence elucidating the role of the CaM antagonist, TFP, in regulating CaM-Fas binding and Fas-mediated DISC formation and apoptosis.Table 1.The change of CaM-Fas binding affinity by the CaM antagonist, TFP. CaM_Fas_WTCaM_1TFP_FasCaM_2TFPs_FasCaM_4TFPs_FasΔG−7.11±4.67−1.32±0.99−6.96±3.47−6.42±1.06ΔΔG 5.79*0.150.69All values in this table were expressed in terms of kcal/mol. ΔG is the binding free energy for the complex, and ΔΔG is the relative binding free energies with respect to the CaM-Fas complex without CaM antagonist TFP(s). * Differences are statistically significant (Student's t-test, p&lt;0.05)
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 96.
Abstract 961: Higher activity of δ-tocopherol (δ-T) than γ-tocopherol and α-tocopherol in the inhibition of lung cancer H1299 xenograft tumors in mice
Authors: Li Guangxun , Picinich Sonia C. , Lee Mao-Jung , Liu Anna Ba , Lu Gary , Yang Chung S. 
Published: February 2011

Abstract
Our previous study showed that γ-TmT (a tocopherol mixture containing 57% γ-T, 24% δ-T, 13% α-T and 1.5% β-T) effectively inhibited colon and lung carcinogenesis as well as the growth of transplanted human lung cancer in mice. This study aims to determine the more active constituents in this mixture in a xenograft model. After injection of human lung cancer H1299 cells into NCr nu/nu nude mice, the mice were maintained on a standard AIN93M diet or diet enriched with α-T, γ-T, δ-T or γ-TmT (each at 0.3% and 0.17%) for 50 days. Dietary δ-T showed the strongest inhibition based on final tumor weight (35.6% and 57.6% reduction from the control by 0.17% and 0.3% δ-T, respectively). Dietary 0.3% γ-T and γ-TmT also showed significant inhibition (37.4% and 46% reduction from control, respectively). However, an inhibitory effect was not observed with dietary α-T group. Consistent with the animal study, the growth of H1299 cells was inhibited by tocopherols with their effectiveness following the order of δ-T &gt; γ-TmT &gt; γ-T, but α-T was not effective. We also found that dietary α-T, γ-T and δ-T treatment respectively increased serum α-T, γ-T and δ-T levels (up to 45, 9.7 and 1.2μM, respectively). However, the serum α-T level was decreased by dietary γ-T and δ-T; for example, at 0.3% they decreased α-T levels by 50% and 42% from the control group, respectively. The results suggest a competition among different forms of tocopherols. Dietary treatment with each form of tocopherol also increased the corresponding side-chain degradation metabolites: carboxyethyl hydroxychromans (CEHC) and carboxymethylbutyl hydroxychromans (CMBHC). With 0.3% α-, γ- and δ-T in the diet, the mean serum levels of α-, γ- and δ-CEHC were 0.08, 0.7 and 3.0μM, respectively; and α-, γ- and δ-CMBHC were 0.14, 0.83 and 4.0μM, respectively. These metabolites retain the intact chromanol ring and, similar to their parent tocopherols, could trap reactive oxygen and nitrogen species. δ-CEHC and δ-CMBHC, which are present at rather high concentrations, may play an important role in inhibiting xenograph and tumor growth. The present results show that δ-T and γ-T effectively inhibited human lung cancer xenograft tumors and the inhibition may be due to the apoptosis-inducing, antioxidative, and reactive nitrogen species-trapping activities of these tocopherols and their metabolites (Supported by NIH grants CA120915, CA122474, and CA133021).
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 961.
Abstract 968: Effects of different types of high fat diets rich in omega-3 or omega-6 fatty acids on uvb-induced complete skin carcinogenesis in skh-1 mice
Authors: Lou You-Rong , Peng Qing-Yun , Li Tao , Medvecky Christopher M. , Wagner George C. , Lu Yao-ping 
Published: February 2011

Abstract
Our previous studies reported that oral administration of caffeine or voluntary exercise may have decreased skin tumor multiplicity, in part, by decreasing fat levels in the dermis and the size of parametrial fat pads. Our studies also showed that surgical removal of the parametrial fat pads (partial lipectomy) enhanced UVB-induced apoptosis in the epidermis. These data suggest that tissue fat may play an important role in regulating UVB-induced apoptosis and skin carcinogenesis. Accordingly, in the present study, we explored the effects of two different types of high fat diets (as described by Rao et al., Cancer Research 61:1927-33, 2001) on UVB-induced complete skin carcinogenesis in SKH-1 hairless mice. Eighty female SKH-1 mice (7-8 weeks old) were equally divided into two groups (40 mice per group). They were irradiated with 30 mJ/cm2 of ultraviolet light B (UVB) once each day, two times per week for 39 weeks. From the beginning of UVB treatment, one group of the mice was given high fat diet containing 32% of polyunsaturated omega-3 fatty acids (HFFO) and the other group of the mice was given high fat diet containing 28% of polyunsaturated omega-6 fatty acids (HFML). Our results showed that although both groups of animals gained weight dramatically, there were no statistical differences on body weight, weight of parametrial fat pad, or thickness of the dermal fat layer. Gross examination showed that HFFO treatment delayed the appearance of UVB-induced skin lesions, decreased the number of mice with tumors and decreased the number and size of tumors per mouse. Histopathological characterization of the skin lesions revealed that HFFO administration decreased the number of squamous cell papillomas per mouse by 64%, decreased the number of keratoacanthomas per mouse by 52% and decreased the number of squamous cell carcinomas per mouse by 46%. In addition to reducing the number of skin tumors, HFFO diet substantially decreased the size of the tumors. That is, the HFFO diet decreased the total volume of squamous cell papillomas per mouse by 98%, decreased the total volume of keratoacanthomas per mouse by 80% and deceased the total volume of squamous cell carcinomas per mouse by 83%. Finally, animals fed the HFFO diet had 22% lower blood levels of tryglycerides (79.3±5.1 mg/dL) than the mice fed the HFML diet (102.0±6.3 mg/dL) (p&lt;0.01), but there was no difference in levels of blood glucose. The effects described here indicate that omega-3 fatty acids in HFFO diet may have beneficial effects against UVB-induced skin carcinogenesis. (Supported by NIH Grant RO1 CA128997)
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 968.
Abstract 973: The NCI Biospecimen Research Network
Authors: Sawyer Sherilyn , Moore Helen M. , Lim Mark David , Vaught Jimmie , Compton Carolyn C. 
Published: February 2011

Abstract
The National Cancer Institute's (NCI) Office of Biorepositories and Biospecimen Research (OBBR) initiated the Biospecimen Research Network (BRN) in early 2006 to coordinate and support systematic investigation into how collection, processing, and storage of human biospecimens affect subsequent analysis. To support this goal, the BRN sponsors, conducts and collaborates on studies to evaluate the effects of biospecimen pre-analytical variables on the outcomes of molecular assays for cancer diagnosis and research. BRN supported findings will contribute to the development of evidence-based best practices for the collection, processing, storage and transport of biospecimens; building on the “NCI Best Practices for Biospecimen Resources.” Currently the BRN sponsors a variety of initiatives aimed to develop and support the research efforts of the biospecimen science community. Through its newly launched extramural research program the BRN supports a portfolio of projects addressing biospecimen pre-analytical variables and biospecimen integrity. Furthermore, the BRN aims to significantly improve biospecimen-based diagnostic and research capabilities by communicating the results of such research to the scientific and clinical communities. The Biospecimen Research Database and the annual BRN symposium, Advancing Cancer Research Through Biospecimen Science are forums established for effective exchange of biospecimen research results. The BRN also supports new research and innovation through its collaborations with other NCI programs such as NCI Innovative Molecular Analysis Technologies (IMAT), the NCI Small Business Innovation Research Development Center (SBIR), the Clinical Proteomics Technologies for Cancer (CPTC) and the Cancer Genome Atlas (TCGA). Through these partnerships, the BRN serves as a resource for program awardees and presents new opportunities for research.
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 973.
Abstract 994: Role of Akt isoforms in regulating intermediate filaments expression in cancer progression
Authors: Fortier Anne-Marie , Cadrin Monique , Asselin Eric 
Published: February 2011

Abstract
Keratins are epithelial-specific intermediate filament (IF) proteins, which are expressed in a tissue-specific manner. As part of the cytoskeleton, keratins are important for the mechanical stability and integrity of epithelial cells and tissues. Moreover, a number of keratins are involved in intracellular signalling pathways which regulate the response to injuries and nonmechanical stresses, cell growth, cell death and cancer progression. Expression of keratins is maintained during malignant transformation of simple epithelial cells. However, vimentin, a mesenchymal-specific IF, is often co-expressed with keratins in late-stage cancer cells presenting a dedifferentiated phenotype. This is associated with an increased metastatic potential. Metastasis, the foremost cause of mortality in cancer patients, is a coordinated biological process involving multiple intracellular signalling pathways. PI3K/Akt pathway has been shown to play a role in invasion/metastasis. Moreover, the three Akt isoforms would play differential roles in this process. In the present study, we investigate the role of Akt isoforms in regulating the expression of keratins and vimentin in different epithelial carcinoma cell lines and the implication of IF in Akt-regulated cell invasion. Our results show that overexpression of constitutively active Akt isoforms up-regulates IF protein and mRNA levels, by increasing IF mRNA stability. On the contrary, down-regulation of Akt isoforms by shRNA decreases IF protein levels. To examine the regulation of IF expression under a more physiological condition, we treated cells with TGF-b1, which is known to induce Akt activation. We demonstrated that IF are up-regulated by TGF-b1 in PI3K-dependent manner. To our knowledge, these results represent the first indication that Akt activity is regulating IF expression. Overexpression of Akt isoforms also increases in vitro cell invasion. Interestingly, we showed that down-regulation of vimentin by siRNA decreases Akt2-induced cell invasion. These results support the hypothesis that modulation of IF expression plays an active role in cancer progression.
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 994.
Abstract 995: Integrin-Linked Kinase localizes in the primary cilia to mediate Hedgehog signaling
Authors: Barakat Badia , Vu Duc , Martellotto Luciano , Watkins David N. , Hannigan Gregory E. 
Published: February 2011

Abstract
The Hedgehog (Hh) signaling pathway is a highly conserved system for regulating metazoan development and cell fate. Integrin linked kinase (ILK) is required for proliferation of Hh-dependent neurons in the mammalian cerebellum, which also give rise to medulloblastoma, the most common childhood brain tumour. Sonic Hedgehog (SHh) binds to the Ptch transmembrane receptor, derepressing the Smo receptor to activate Hh signaling and key downstream targets such as Gli transcription factors. We find that SHh-induced osteogenic differentiation of C3H10T1/2 cells is blocked (&gt;90% inhibition) by a small molecule ILK inhibitor, or by ILK-specific RNAi. SHh induces alkaline phosphatase (AP) activity in control 10T1/2 cells, but not cells transfected with ILK siRNA (&gt;75% knockdown). Moreover we find that the small molecule ILK inhibitor suppresses SHh-induced differentiation in a dose-dependent manner, as effectively and to the same extent as the well-characterized selective Hh inhibitor, cyclopamine. Activation of Smo with the agonist, SAG, induces ILK-dependent AP induction in these cells, suggesting ILK acts downstream of Smo in the Hh pathway. SHh induction of the Gli1 transcription factor is also blocked by inhibiting ILK, suggesting ILK acts upstream of direct Hh targets. ILK inhibitor or siRNA suppresses constitutively activated Gli1 in medulloblastoma cells, derived from Ptch+/-:p53 compound heterozygote mice. In support of ILK involvment in Hh signaling, we have discovered a novel localization of ILK that indicates spatial proximity to Hh signaling molecules. The primary cilium is a microtubule-based organelle that is required for Hedgehog signaling, and we now find that ILK localizes in this structure. Using a selective ciliary marker, acetylated α-tubulin, we find that ILK localizes in the primary cilium along with components of the Hh pathway. Inhibition of ILK does not inhibit cilia formation, indicating that ILK is not involved in ciliogenesis. IFT proteins (eg. kinesin II motor subunit Kif3a), regulate both the assembly of, and the entry of Hh signaling proteins to, the cilium. Knockdown of Kif3a inhibits ciliogenesis and efficiently blocks SHh signaling, strongly suggesting the ciliary pool of ILK is critical for Hh signaling. Our work points to ILK as a novel therapeutic target in Hh-activated cancers.
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 995.
Abstract LB-112: The human papillomavirus type 16 E5 protein represses X-box binding protein 1 and cyclooxygenase-2 stress signaling in keratinocytes
Authors: Sudarshan Sawali R. , Liu Xuefeng , Schlegel Richard 
Published: February 2011

Abstract
Human papillomavirus (HPV) is the causative agent of 99% of cervical cancers, around 60% of which are caused by the high-risk type 16. While two of its three oncoproteins, E6 and E7, have been well characterized, the biological functions of the E5 protein remain elusive. Recent studies have suggested that E5 enhances HPV-induced oncogenesis, although in isolation it is not tumorigenic. In this study, we analyzed the gene expression changes induced by the endoplasmic reticulum (ER) resident HPV-16 E5 protein in primary human keratinocytes. Using real-time RT-PCR, we identified the downregulation of spliced X-box binding protein 1 (XBP-1), a key player in the ER-stress response, as a biochemical marker of E5 expression. The transmembrane kinase receptor responsible for the cleavage of XBP-1, ER-to-nucleus signaling 1 protein (IRE1α/ERN1), was also downregulated in E5-transduced cells. Furthermore, a microarray analysis of E5-transduced keratinocytes revealed the downregulation of another member of the ER-stress response pathway, prostaglandin G/H synthase and cyclooxygenase (COX-2/PTGS2). In addition, levels of COX-2 and XBP-1 were measured in keratinocytes expressing either low-risk HPV-6b E5 or a C-terminal 16E5 mutant containing a two amino acid mutation. The latter mutant had the histidine and alanine residues (conserved in the high-risk HPVs) replaced with tyrosine and isoleucine (conserved in the low-risk HPVs). Importantly, neither 6bE5 nor the H77YA78I 16E5 mutant was able to affect levels of these ER-stress response genes, suggesting that the ability to induce such changes in gene expression is specific to high-risk 16E5. Since preliminary data showed that expression of the E6 and E7 proteins in keratinocytes greatly induced XBP-1 splicing, we hypothesized that 16E5 might play a protective role against the host response to viral infection. In agreement with this postulate, we found that cells co-expressing E6, E7 and E5 had lower levels of COX-2 expression than cells expressing E6 and E7 alone. Interestingly, the levels of COX-2 in E6/E7-transduced cells were also supressed when compared to vector alone, suggesting that multiple pathways of COX-2 repression may exist. The mechanism by which E5 downregulates COX-2 is currently unknown, but it can be overcome by treatment with thapsigargin or tunicamycin, which initiate ER stress via calcium fluxes and abnormal protein glycosylation respectively. It is therefore unlikely that E5 is specifically tempering these pathways. Our data suggest a potential role for E5 in repressing the cellular ER-stress response following HPV infection.
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 LB-112.
Abstract LB-118: 17AAG, through hydrophobic binding, targets mitochondrial VDAC and inhibits cell invasion
Authors: Xie Qian , Wondergem Robert , Shen Yuehai , Cavey Greg , Ke Jiyuan , Thompson Ryan , Bradley Robert , Xu Yong , Holtrop Jennifer , Omar Hanan , Wenkert David , Xu Eric , Vande Woude George F. 
Published: February 2011

Abstract
17-allylamino-17-demethoxygeldanamycin (17AAG) is a member of the geldanamycin (GA) benzoquinone ansamycin family of anticancer drugs that is in clinical trials against several types of cancer. GA compounds bind to the ATP pocket of HSP90, and block its chaperone function resulting in the degradation of a wide variety of substrate oncoproteins, thereby providing a powerful “multi-target therapy”. GA compounds, however, are highly hydrophobic, requiring complicated drug vehicle formulation. In attempting to understand the influence of the hydrophobicity of GA drugs in vitro, we discovered that the hydrophobic molecules, GA or 17AAG, can bind directly to the highly hydrophobic voltage-dependent anion channel (VDAC) or to purified mitochondria where VDAC resides. Moreover, 17AAG binds to purified mitochondria prepared from all tissues and cells tested and compete with HSP90. Consistent with benzoquinone-ubiquinones functioning as classical Ca2+ mitochondrial regulators we associated the benzoquinone ring which all members of the GA family of drugs have as the responsible GA binding moiety with mitochondria. We show that both the ubiquinone, UbO, and GA drugs increase intracellular Ca2+, most likely through intracellular mitochondrial stores and both drugs diminish the plasma cationic current. By contrast, radicicol, with only an ansamycin ring, binds tightly to HSP90, but lacking the benzoquinone ring, has no measurable effect on cationic current or intercellular Ca2+ concentration. Thus, we propose that benzoquinone mitochondria binding through hydrophobic VDAC represents an alternative mode of action of GA drugs that can either alter HSP90 activity or be responsible for toxicity. Both activities must be considered when novel GA derivatives are developed as therapeutics.
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 LB-118.
Abstract LB-119: Production of functionally active glycoproteins and phosphoproteins based on a novel human in vitro expression system
Authors: Kilmer Greg , Vattem Krishna , Hommema Eric , Maas Ken , Deshpande Atul , Webb Brian L. 
Published: February 2011

Abstract
One of the key features of the human proteome is the abundance of post-translation modification (PTM), with more than 50% of proteins being glycosylated and more than 30% of all the proteins regulated by kinase activities. The alteration of protein glycosylation and other PTMs such as phosphorylation in cancer cells is closely related to the onset and progression of cancer and other disease states. In collaboration with RIKEN, we have developed a novel human in vitro translation (also known as cell-free expression) system, that can produce biologically active glycoproteins such as human chorionogonadotrophic hormone as well as active protein kinases such as Akt, MAPK and Aurora A kinase. A highly efficient PCR based approach will also be discussed that eliminates the need for cloning of genes into the expression vector and provides an easier expression of genes present in any other vector. In addition we demonstrate that our in vitro human expression, unlike the only other commercially existing mammalian expression system based on rabbit reticulocytes, can sustain expression of proteins for over 6 hours without the need to remove of inhibitory by products and is compatible with fluorimetric and colorimetric assays.
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 LB-119.