bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3187: Genomic abnormalities in air pollution-related lung cancer
Authors: Zhou Guang-Biao , Yu Xian-Jun , Wu Li-Chuan , Huang Yun-Chao , Li Gao-Feng , Wen Zhe-Sheng , Chen Sai-Juan , Cao Yi , Wang Sheng-Yue , Yang Min-Jun , Chen Zhu
Published: May 2015

The lung cancer incidence in Xuanwei (XW) City and Fuyuan (FY) County in Yunnan Province of China is among the highest in China, owing to high levels of air pollution produced by combustion of smoky coal. This epidemiology profile provides a unique opportunity for identifying the carcinogenic mechanism underlying air pollution. Here, we analyze the genome sequences of 14 XW/FY non-small-cell lung cancers and their paired normal tissues, and report a mean of 289 somatic exonic mutations and 71 genomic rearrangements/tumor. Among the recurring mutated genes, calcium signaling and ion channel genes are the most frequently altered gene category. Higher mutation rate in calcium signaling genes is confirmed by genomic resequencing in 65 XW/FY patients and 85 patients from control regions. Small interfering RNA mediated silencing shows that inactivation of calcium channel leads to inhibition of lung cancer cells, suggesting a role for calcium signaling molecules in air pollution-related carcinogenesis leading to lung cancer.

Citation Format: Guang-Biao Zhou, Xian-Jun Yu, Li-Chuan Wu, Yun-Chao Huang, Gao-Feng Li, Zhe-Sheng Wen, Sai-Juan Chen, Yi Cao, Sheng-Yue Wang, Min-Jun Yang, Zhu Chen. Genomic abnormalities in air pollution-related lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3187. doi:10.1158/1538-7445.AM2014-3187
Abstract 3188: Evaluation of clonal origin of malignant mesothelioma/polyclonal origin of malignant mesothelioma
Authors: Comertpay Sabahattin , Mezzapelle Rosanna , Tanji Mika , Strianese Oriana , Pass Harvey I. , Weigel Tracey , Friedberg Joseph , Sugarbaker Paul , Krausz Thomas , Wang Ena , Gaudino Giovanni , Yang Haining , Powers Amy , Parsons Barbara , Pastorino Sandra , Carbone Michele
Published: May 2015

Tumors are generally thought to be monoclonal in origin, although this paradigm stemmed decades ago mostly from the study of hematopoietic malignancies and sarcomas. The clonal origin of malignant mesothelioma, a deadly cancer resistant to the current therapies, has never been investigated before. We used the HUMARA (human androgen receptor) assay to examine the pattern of clonality in 14 sporadic and 2 familial Malignant Mesotheliomas (MM). Of 16 specimens studied, 14 were informative and nearly all (13/14) revealed two electrophoretically distinct methylated HUMARA alleles, indicating a polyclonal origin for these tumors. This discovery has important clinical implications, in that an accurate assessment of tumor clonality is key to the design of novel molecular strategies for the treatment of MM.

Citation Format: Sabahattin Comertpay, Rosanna Mezzapelle, Mika Tanji, Oriana Strianese, Harvey I. Pass, Tracey Weigel, Joseph Friedberg, Paul Sugarbaker, Thomas Krausz, Ena Wang, Giovanni Gaudino, Haining Yang, Amy Powers, Barbara Parsons, Sandra Pastorino, Michele Carbone. Evaluation of clonal origin of malignant mesothelioma/polyclonal origin of malignant mesothelioma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3188. doi:10.1158/1538-7445.AM2014-3188
Abstract 3189: Morphological and molecular study of cell-chrysotile interaction in two different cell lines
Authors: Ricardi Luana R. , Rezende-Teixeira Paula , Souza Marcelo Medina de , Machado-Santelli Glaucia M.
Published: May 2015

Chrysotile accounts for more than 90% of the asbestos used worldwide. This type of fiber is considered the least oncogenic among the asbestos fibers, mainly based on the fiber dimension and biopersistence inside the lung. It is associated with respiratory diseases and lung cancer and identified as human carcinogen by the International Agency for Research on Cancer. However the mechanism and potential of different fibers to cause cancer is still a matter of debate. The present study focuses the interaction of fibers with different cell types, analyzing their ability and mechanism of fiber internalization. After short chrysotile small fibers exposure time (4-8hrs) both non-small cell lung carcinoma and monocytic cell lines were able of fiber internalization. Transmission electron microscope (TEM) analyses showed fibers involved or not by membrane into the cytoplasm and sometimes seemed to be associated with the chromatin. Cytoskeleton elements were observed to be concentrated around the fibers and they were identified as intermediate filaments according to their diameter. Immunofluorescent preparations imaged by laser scanning confocal microscope and submitted to localization analyses showed that both vimentin and cytokeratin may participate in this process. The data suggest that the internalization of fibers may occur through endocytosis and/or by physical perforation of the plasma membrane.

Since the scavenger receptors are responsible for recognition and internalization of dust particles by macrophages in lung, and some of them are linked to silica-containing particles recognition we evaluated their expression level of mRNA by using quantitative RT-PCR. The scavenger receptors appear to interfere with the internalization of chrysotile fibers process. SR-A and SCARA5 change their relative expression of mRNA after cell exposure to chrysotile fibers, such as SR-A and SCARA5. Furthermore, our study was the first to find the relationship between the SCARA5 receptor and chrysotile fibers. To check if features of the chrysotile internalization by epithelial and monocytic cells would be related with fibers shape or chemical composition, the cells were exposed to single walled carbon nanotubes in similar conditions.

Financial support: FAPESP, CAPES and CNPq)

Citation Format: Luana R. Ricardi, Paula Rezende-Teixeira, Marcelo Medina de Souza, Glaucia M. Machado-Santelli. Morphological and molecular study of cell-chrysotile interaction in two different cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3189. doi:10.1158/1538-7445.AM2014-3189
Abstract 3191: Elevation of tumor-promoting cytokines in mice exposed to the environmental contaminant tributyltin
Authors: Lawrence Shanieek , Pellom Samuel Troy , Hurd-Brown Tasia , Shanker Anil , Whalen Margaret
Published: May 2015

Cytokines are important regulators of immune responsiveness. Alterations of cytokine secretion from immune cells would have the potential to disrupt both immune destruction of cancer cells and to alter the proliferation and invasiveness of tumor cells due to their capacity to act as growth factors, angiogenic factors, and inducers of epithelial-mesenchymal transition. Tributyltin (TBT) is a widespread environmental contaminant having been used in a variety of industrial applications and is found in human blood at levels ranging as high as 261 nM. Studies in human immune cells have shown that secretion of tumor necrosis factor alpha (TNFα) is altered by exposures to TBT. There have been no in vivo studies of the effects of exposure to TBT on the cytokine communication network. As it is not possible to examine alterations of this network in exposed humans, we examined the effects of TBT exposure in a mouse model using Milliplex mouse cytokine/chemokine magnetic bead premixed 32 plex kit. Serum and spleens of TBT-exposed mice were analyzed for changes in cytokine secretion/levels. TBT increased the levels of IL13 in the serum of non-tumor-bearing mice and increased IL1β, IL6, KC, and M-CSF in tumor-bearing mice. The spleens of both non-tumor-bearing and tumor-bearing mice showed increases in Eotaxin when exposed to TBT. Of significant interest is the fact that the presence of tumor causes increases in Eotaxin, IL12p40, M-CSF, and TNFα and exposure to TBT alone can cause increases in these same cytokines. Tumors are known to increase levels of cytokines that enhance their ability to survive. The results showing that TBT can amplify the levels of several cytokines that are increased by tumor suggest that TBT may have tumor-promoting abilities by modulating tumor growth and/or invasiveness. Experiments are under way to dissect cytokine signaling pathways that are affected by TBT. The findings will allow identification of molecular targets that are necessary to intercept the tumor promoting signaling following TBT exposure.

Supported by: NIH grant 5U54CA163066-03

Citation Format: Shanieek Lawrence, Samuel Troy Pellom, Tasia Hurd-Brown, Anil Shanker, Margaret Whalen. Elevation of tumor-promoting cytokines in mice exposed to the environmental contaminant tributyltin. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3191. doi:10.1158/1538-7445.AM2014-3191
Abstract 3194: Platycodin D induces growth arrest and cell cycle arrest by activating MDM2 and interacting with FOXO3a in prostate cancer
Authors: Xu Hongxia , Zhou Rui , Lu Zongliang , Liu Kai
Published: May 2015

Purpose: The present study was designed to evaluate the anti-tumor activity and possible mechanism of action of Platycodin D (PD), a major saponin derived from Platycodin grandiflorum, in human prostate cancer.

Experimental Design: Human prostate cancer cells with different p53 status (p53 wildtype LNCaP cells, p53 null PC3 cells and p53 mutant DU145 cells), and the normal human prostate cell line, RWPE-1, were exposed to various concentrations of PD to compare its cytotoxicity in vitro (via the MTT assay). Detailed invitro and invivo studies on the anti-tumor effects of PD were then performed.

Results: PD exerted cytotoxicity against all three prostate cancer cell lines, with half-maximal inhibitory concentrations in the range of 11.16 to 26.13μmol/L. The PC3 cells were the most sensitive to PD, whereas RWPE-1 cells were not sensitive to PD.A further study in these cell lines showed that PD could potently affect the cell proliferation (indicated by the bromodeoxyuridine assay), induce cell apoptosis (by Annexin V-FITC flow cytometry) and cause cell cycle arrest (PI staining). After being treated with PD for 48 hours,the DU145 and LNCaP cells were arrested in the G0 /G1 phase, and the PC3 cells were arrested in the G2/M phase. A Western blot analysis indicated that PD decreased the expression of MDM2 and increased the expression levels of FOXO3a, p21 and p27 in the three cell lines. In PC3 cells, PD exposure decreased the levels of cell cycle-related proteins, including E2F1, cdk2, cdk4, cdk6, cyclin D1 and cdc2. PD exposure increased apoptosis-related proteins, Bax, caspase9 and caspase3, and downregulated Bcl-2, PARP, and caspase8. Real-time PCR indicated that the same changes had occurred at the transcriptional level for all of these targets. Moreover, PD dose-dependently inhibited the growth of PC3 xenograft tumors in BALB/c nude mice. A Western blot analysis of the xenograft tumors indicated that similar changes in protein expression also occurred in vivo.

Conclusion: PD exhibits significant activity against prostate cancer cells in a p53-independent manner. MDM2 and the FOXO3a transcription factor appear to be involved in the mechanism of action of PD, which provides a basis for future development of this agent for human prostate cancer therapy.

Citation Format: Hongxia Xu, Rui Zhou, Zongliang Lu, Kai Liu. Platycodin D induces growth arrest and cell cycle arrest by activating MDM2 and interacting with FOXO3a in prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3194. doi:10.1158/1538-7445.AM2014-3194
Abstract 3196: Antitumor effect of a Compound A, a derivative of a naturally occurring rotenoid deguelin, by inhibition of ATP binding to heat shock protein 90
Authors: Lee Su-Chan , Lee Ju-Sung , Hyun Seung-Yeob , Choi Hoon , An Hongchan , Kim Kyu-Won , Suh Young-Ger , Lee Ho-Young
Published: May 2015

Despite several decades of intensive efforts to develop anti-cancer drugs and therapeutic strategies, lung cancer is still one of the main causes of cancer-related deaths worldwide. Deguelin, a natural plant product, has shown anti-cancer activities in skin, colon, breast and non-small cell lung (NSCLC) cancers. However, the potential side effects of deguelin could be an obstacle to its use in clinic. In the current study, we aimed to develop a derivative of deguelin with reduced potential side effects. We have synthesized a novel deguelin analog, compound A, and evaluated its effects on cancer and normal bronchial epithelial cell proliferation, survival, and colony formation by the MTT, FACS, and/or anchorage-dependent and -independent colony forming assays. Mechanisms underlying anti-tumor effects of compound A were investigated by Western blot, and RT-PCR. Treatment with compound A inhibited cancer cell proliferation and colony formation, promoted cell cycle arrest and induced apoptosis. Compound A also suppressed the expression of HIF-1α protein and its target gene, VEGF, in NSCLC cells. Administration of compound A to NSCLC xenograft-bearing mice significantly decreased tumor growth by inhibiting tumor angiogenesis and by inducing apoptosis. In its action mechanism, compound A was involved in binding to the ATP-binding pocket of HSP90, leading to the suppression of HSP90 function. These collective results suggest that a novel deguelin derivative, compound A, has a strong potential as an anti-cancer drug.

Citation Format: Su-Chan Lee, Ju-Sung Lee, Seung-Yeob Hyun, Hoon Choi, Hongchan An, Kyu-Won Kim, Young-Ger Suh, Ho-Young Lee. Antitumor effect of a Compound A, a derivative of a naturally occurring rotenoid deguelin, by inhibition of ATP binding to heat shock protein 90. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3196. doi:10.1158/1538-7445.AM2014-3196
Abstract 3197: Gedunin inactivates the co-chaperone p23 causing cancer cell death by apoptosis
Authors: Patwardhan Chaitanya A. , Peterson Laura B. , Blagg Brian , Chadli Ahmed
Published: May 2015

Pharmacological inhibition of Hsp90 is an exciting option for cancer therapy. The clinical efficacy of Hsp90 inhibitors is, however, less than expected. Binding of the co-chaperone p23 to Hsp90, and induced overexpression of anti-apoptotic proteins, Hsp70 and Hsp27, are thought to contribute to this outcome. Herein, we report that the natural product, gedunin, may provide a new alternative to inactivate the Hsp90 machine. We show that gedunin directly binds to p23 and inactivates it, without overexpression of Hsp27 and relatively modest induction of Hsp70. Using molecular docking and mutational analysis, we mapped the gedunin-binding site on p23. Functional analysis shows that gedunin inhibits p23 chaperoning activity, blocks its cellular interaction with Hsp90 and interferes with p23-mediated gene regulation. Cell treatment with gedunin leads to cancer cell death by apoptosis through inactivation of p23 and activation of caspase 7, which cleaves p23 at the C-terminus. These results provide important insight into the molecular mechanism of action of this promising lead compound.

Citation Format: Chaitanya A. Patwardhan, Laura B. Peterson, Brian Blagg, Ahmed Chadli. Gedunin inactivates the co-chaperone p23 causing cancer cell death by apoptosis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3197. doi:10.1158/1538-7445.AM2014-3197
Abstract 3199: Pterostilbene supresses prostate cancer progression in transgenic mice model
Authors: Zhang Liangfen , Dhar Swati , Rimando Agnes M. , Kumar Avinash , Lage Janice , Lewin Jack R. , Zhang Xu , Levenson Anait S.
Published: May 2015

Metastasis-associated protein 1 (MTA1) is a cancer progression-related epigenetic modifier whose overexpression is associated with aggressive prostate cancer (PCa), disease recurrence and metastasis in men. We previously found that MTA1 expression is significantly increased in Pten-null murine cells, and that resveratrol and its analogues inhibit MTA1 expression. In the current study, we used the murine Pten knockout PCa model to evaluate the effect of pterostilbene (PTER) on MTA1-mediated events in PCa progression. Pterostilbene is a potent natural analogue of resveratrol with improved pharmacokinetic properties and superior pharmacological potency. The Pten conditional knockout mice that mimic human disease and provide fast stage-defined development of PCa were fed, starting at 3 weeks of age, with phytoestrogen-free diet AIN 76A and injected daily (i.p.) either with 10% DMSO (control, untreated) or with 10 mg/kg bw PTER until 6, 10, 15 and 25 weeks of age. At necropsy, mouse prostates were formalin-fixed and paraffin-embedded for histological and immunohistochemical analysis. For protein analysis, mouse prostate dissection was performed to distinguish anterior prostate (AP) and dorso-latero-ventral (DLV) lobes. Our results showed that the PTER-treated group had a lower tumor grade and higher incidence of prostate intraepithelial neoplasia (PIN) at the expense of a substantial decrease in adenocarcinoma incidence. Prostate tissues from the PTER-treated group presented a decrese in Ki67 proliferation marker-positive cells and an increase in apoptosis. As potential mechanisms of PTER efficacy, 50-75% decrease in MTA1 and pAkt levels were observed at different time points. In addition, PTER inhibited androgen receptor (AR) expression, especially in the AP. Taken together, our findings suggest that PTER blocks PCa growth and progression at PIN stage in Pten conditional knockout mice model through inhibition of MTA1-Akt axis.

Citation Format: Liangfen Zhang, Swati Dhar, Agnes M. Rimando, Avinash Kumar, Janice Lage, Jack R. Lewin, Xu Zhang, Anait S. Levenson. Pterostilbene supresses prostate cancer progression in transgenic mice model. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3199. doi:10.1158/1538-7445.AM2014-3199
Abstract 320: Frequent inactivation of the Mieap-regulated mitochondrial quality control in pancreatic and breast cancer
Authors: Kamino Hiroki , Nakamura Yasuyuki , Sano Hitoya , Murai Ryuya , Saito Yuri , Futamura Manabu , Yoshida Kazuhiro , Hiraoka Nobuyoshi , Kanai Yae , Kushima Ryoji , Katagiri Toyomasa , Arakawa Hirofumi
Published: May 2015

Mieap, a p53-inducible protein, controls mitochondrial quality by repairing or eliminating unhealthy mitochondria via MALM (Mieap-induced Accumulation of Lysosome-like organelles within Mitochondria) or MIV (Mieap-Induced Vacuole), respectively (1, 2). Two mitochondrial outer membrane proteins, BNIP3 and NIX are essential mediators of MALM (3). Previously, we reported that the Mieap-regulated mitochondrial quality control (MQC) pathway was inactivated in almost 80% of colorectal cancer patients. To evaluate the role of the Mieap-regulated MQC in pancreatic and breast cancer, we analyzed the status of p53, Mieap, BNIP3 and NIX in 50 cases of primary pancreatic cancer and 63 cases of primary breast cancer by examining p53 mutation and Mieap/BNIP3/NIX promoter methylation. As the result, the Mieap-regulated MQC pathway was inactivated in 71% of pancreatic and 27% of breast cancer patients. Interestingly, the BNIP3 promoter methylation was frequently detected in colorectal and pancreatic cancer patients (49% and 58%, respectively) whereas it was not detected in breast cancer patients at all (0%). Instead, the Mieap promoter methylation was highly detected in breast cancer patients (12.7%) compared to colorectal and pancreatic cancer patients (8.8% and 2.0%, respectively). These results suggest that the Mieap-regulated MQC pathway plays a critical role in tumorigenesis of pancreatic and breast cancer, and that there may be some different mechanisms between colorectal/pancreatic and breast cancers for inactivation of the Mieap-regulated MQC pathway. Inactivation of Mieap-regulated MQC pathway leads to the accumulation of unhealthy mitochondria generating high level of mitochondrial reactive oxygen species. We are now investigating whether the p53/Mieap/BNIP3 MQC pathway affects cell growth, cell motility or tumorigenicity using several pancreatic and breast cancer cell lines that are subcutaneously implanted into nude mice. In this paper, we discuss the potential role of the Mieap-regulated MQC in pancreatic and breast tumor suppression.

1. Miyamoto Y et al. PLoS ONE 6: e16054, 2011

2. Kitamura N et al. PLoS ONE 6: e16060, 2011

3. Nakamura Y et al. PLoS ONE 7: e30767, 2012

Citation Format: Hiroki Kamino, Yasuyuki Nakamura, Hitoya Sano, Ryuya Murai, Yuri Saito, Manabu Futamura, Kazuhiro Yoshida, Nobuyoshi Hiraoka, Yae Kanai, Ryoji Kushima, Toyomasa Katagiri, Hirofumi Arakawa. Frequent inactivation of the Mieap-regulated mitochondrial quality control in pancreatic and breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 320. doi:10.1158/1538-7445.AM2014-320
Abstract 3201: Parthenolide ameliorates experimental colitis associated colon cancer by inhibition of NF-kB signaling
Authors: Kim Se Lim , Kim Sang-Wook , Kim Seong Hun , Kim In Hee , Lee Seung Ok , Lee Soo Teik , Kim Dae Ghon
Published: May 2015

Parthenolide (PT), a NF-kB inhibitor, has recently been demonstrated as a promising anticancer agent that promotes apoptosis of cancer cells. We previously showed that PT suppresses tumor growth in a xenograft model of colorectal cancer cells by regulation of Bcl-2 family. Unfortunately, little is known about its role in the process of tumor development in colitis associated colon cancer (CAC). Therefore, this study was designed to investigate the effects of PT on an experimental murine CAC model. Experimental CAC was induced by azoxymethane (AOM) and dextran sulfate sodium (DSS). Mice were divided into 3 groups: AOM+DSS, AOM+DSS+2mg/kg PT and AOM+DSS+4mg/kg PT. We demonstrated that administration of PT significantly reduced the severity of AOM/DSS-induced CAC as assessed by histological analysis, and resulted in downregulation of phospho-NF-κB p65 expression by the blockade of phosphorylation and subsequent degradation of IκB-a. Administration of PT ameliorated the carcinogenesis through the downregulation of antiapoptotic protein Bcl-2 and Bcl-xL mediated by inhibition of NF-kB activation. Moreover, apoptosis and caspase-3 expression also increased markedly in PT administration group. These findings demonstrate that PT downregulates NF-kB resulting in initiation of apoptosis and eventual suppression of CAC development, suggesting that PT exerts beneficial effects in experimental CAC and could therefore be a potential chemopreventive and therapeutic agent of CAC.

Citation Format: Se Lim Kim, Sang-Wook Kim, Seong Hun Kim, In Hee Kim, Seung Ok Lee, Soo Teik Lee, Dae Ghon Kim. Parthenolide ameliorates experimental colitis associated colon cancer by inhibition of NF-kB signaling. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3201. doi:10.1158/1538-7445.AM2014-3201
Abstract 3202: Deciphering the targets and mechanism of action of the natural product Tonantzitlolone in clear cell renal cell carcinomas
Authors: Sourbier Carole , Scroggins Brad , Mannes Philip Z. , Beutler John A. , Linehan W. Marston , Neckers Len
Published: May 2015

Tonantzitlolone is a diterpene ester first extracted in 1990 from the native Mexican plant Stillingia sanguinolutea (Euphorbiaceae), which displays anti-tumor activity. Although its structure and total synthesis have been elucidated, its cellular target(s) and potential mechanism of action remain unknown. We identified Tonantzitlolone as a dual PKCα and PKCθ activator in vitro. However in clear cell renal cell carcinoma (CCRCC), its activity was mostly PKCθ-dependent. Through activation of PKCθ, Tonantzitlolone induced an insulin-resistant phenotype by inhibiting IRS1 and the PI3K/Akt pathway. Simultaneously, Tonantzitlolone activated the transcription factor heat shock factor 1 (HSF1), driving glucose dependency. Thus, similar to the selective PKCθ activator Englerin A, Tonantzitlolone induces a metabolic catastrophe in CCRCC, starving the cells of glucose while increasing their glycolytic dependency, ultimately leading to cell death.

Citation Format: Carole Sourbier, Brad Scroggins, Philip Z. Mannes, John A. Beutler, W. Marston Linehan, Len Neckers. Deciphering the targets and mechanism of action of the natural product Tonantzitlolone in clear cell renal cell carcinomas. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3202. doi:10.1158/1538-7445.AM2014-3202
Abstract 3203: Honokiol inhibits growth and migration of renal cell carcinoma
Authors: Cheng Shujie , Welty Matt , Eliaz Isaac , Castillo Victor , Sliva Daniel
Published: May 2015

Renal cell carcinoma (RCC) is a common urological cancer worldwide and is known to the high risk of recurrence and metastasis. Approximately 70% of patients with RCC will develop recurrence after surgical resection, and 25%-30% of patients will eventually develop progression to distant metastasis. Honokiol is a small-molecule polyphenol isolated from the genus Magnolia, which has been shown to be a potential anticancer agent in multiple facets of signal transduction. Here we demonstrate that honokiol inhibits proliferation of RCC cells 786-0 and A498 without affecting cell viability. Moreover, honokiol also significantly inhibited migration of 786-0 cells in a dose-dependent manner. DNA microarray analysis showed that honokiol regulated expression of many genes related to human tumor metastasis in 786-0 cells. Real time PCR analysis confirmed that the expression of metastasis suppressor KISS1 and its receptor, KISS1R, were upregulated in 786-0 cells after treatment with honokiol. In addition, the shape changes and excessive formation of actin stress fibers were identified in 786-0 cells treated with honokiol. This phenomenon disappeared when treated cells with the pharmacological Rho-kinase inhibitor Y-27632 and honokiol. This inhibition can also be identified in 786-0 cells treated with Y-27632 only. Our present results demonstrated that honokiol could inhibit the growth and migration of RCC, which is likely to be regulated by the Rho and Rho-Associated Kinase (ROCK) pathway. In conclusion, honokiol is a biologically active natural compound which can be considered for the alternative treatment of RCC. The investigation of detailed mechanisms and molecular targets are in progress.

Citation Format: Shujie Cheng, Matt Welty, Isaac Eliaz, Victor Castillo, Daniel Sliva. Honokiol inhibits growth and migration of renal cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3203. doi:10.1158/1538-7445.AM2014-3203
Abstract 3207: Curcumin induces pancreatic cancer cell death by targeting IAPs
Authors: Osterman Carlos J. Diaz , Valenzuela Malyn May Asuncion , Bennit Heather R. Ferguson , Khan Salma , Wall Nathan R.
Published: May 2015

Background

Pancreatic cancer has among the highest mortality rates compared to other cancer types, with 94% of patients dying within five years of diagnosis. Contributing to the high mortality rate, patients with pancreatic cancer do not present significant symptoms until the disease is advanced. Moreover, treatment strategies have not improved substantially over the last decade. For this reason, a recent trend has led toward focusing on the use of herb-derived compounds, such as curcumin (turmeric derivative), to explore alternative treatments that will improve current pancreatic cancer therapy.

Objective

The objectives of this study were (1) to determine the effects of curcumin on pancreatic cancer cell metabolism, viability, cell cycle and morphology, and (2) to determine the mechanism by which curcumin induces cell death in pancreatic cancer cells by analyzing intracellular expression levels of inhibitor of apoptosis proteins (IAPs).

Design/method

Pancreatic adenocarcinoma cell lines (Panc-1 and MiaPaCa-2) were used to assess the effects of curcumin in pancreatic cancer. Effects on metabolism, viability and cell cycle were assayed by Alamar Blue, Trypan Blue and Propidium Iodide (PI) staining/flow cytometry (FACSCalibur). Morphological imaging was performed using Hoffman Modulation microscopy. Apoptosis was detected by Annexin V/PI flow cytometry (MACSQuant) and the expression of IAPs was assessed by Western blot.

Results

These data show that curcumin decreases pancreatic cancer cell metabolism and viability. Cell cycle stages were arrested in pancreatic cancer cells treated with curcumin compared to the non-treated controls. Apoptotic cells were detected by microscopy and confirmed by Annexin V/PI staining with subsequent flow cytometry analysis. IAPs levels are reduced in pancreatic cancer cell lines treated with curcumin compared to non-treated controls.

Conclusion

These results demonstrate that pancreatic adenocarcinoma cell lines are sensitive to curcumin treatment. Moreover, curcumin's possible mechanism of action is inducing apoptosis by targeting IAPs. Collectively, these data suggest a potential role for curcumin in pancreatic cancer therapy.

Citation Format: Carlos J. Diaz Osterman, Malyn May Asuncion Valenzuela, Heather R. Ferguson Bennit, Salma Khan, Nathan R. Wall. Curcumin induces pancreatic cancer cell death by targeting IAPs. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3207. doi:10.1158/1538-7445.AM2014-3207
Abstract 3208: Identification and characterization of a small molecule involved in the downregulation of RET transcription in MTC
Authors: Kumarasamy Vishnu Muthuraj , Shin Yoon-Joo , Sun Daekyu
Published: May 2015

The gain of function mutations in the RET proto-oncogene, which results in the constitutive activation of the mitogenic pathways, were found mainly in Medullary Thyroid Carcinoma (MTC). In our study, a new therapeutic strategy has been explored to repress the expression of the RET gene by targeting the transcriptional activation of this gene with small molecules. The transcriptional inhibitor of the RET gene was initially screened from the NCI chemical libraries using the cell-based luciferase reporter gene assay, which has a promoter sequence identical to the RET promoter. As a result of our screening effort, a small molecule, nonactin, has been identified as a potential lead compound, downregulating the transcriptional activity of the RET gene promoter. The effect of this molecule in the downregulation of the RET mRNA levels and the protein levels was confirmed by RT-PCR and western blotting respectively. We also observed that Egr1 expression is downergulated in a dose dependent manner by this small molecule, which is in correlation with the RET downregulation. The RET promoter has a highly conserved GC rich region, which acts as a binding site for Egr-1, which is involved in the transcription of many growth factor genes. Thus, we conclude that Egr1 would be one of important transcriptional factors involved in the activation of the RET and nonactin inhibits the expression of Egr1, which in turn downregulates the transcription of RET oncogene.

Citation Format: Vishnu Muthuraj Kumarasamy, Yoon-Joo Shin, Daekyu Sun. Identification and characterization of a small molecule involved in the downregulation of RET transcription in MTC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3208. doi:10.1158/1538-7445.AM2014-3208
Abstract 3214: C-seco taxanes with 7, 9-linkages exhibit better activity in drug-resistant tumor cells
Authors: Tang Yong , Cai Pei , Rodríguez-Salarisch Javier , Díaz J. Fernando , Fang Weishuo
Published: May 2015

Taxanes are a group of very successful MT stabilizing agents in cancer chemotherapy, although they suffer from tumor drug resistance, which compromises clinical efficacy. Many efforts were made in our labs to the structural modification of paclitaxel to improve efficacy against tumor drug resistance mediated by multiple mechanisms, with the emphasis on two clinically relevant ones: P-glycoprotein (Pgp) and β-III tubulin overexpression. While Pgp mediated multidrug resistance has been extensively explored and applied to the drug discovery projects, β-III tubulin overexpression related drug resistance was much less explored.

The β-III tubulin isotype was found to play an important role in conferring resistance to MT stabilizing agents only in recent decade. It has also been identified as a survival factor in several types of cancers, thus conferring resistance to various anticancer agents, such as MT destabilizing agents and DNA interacting agents.

We have found some high affinity taxanes (e.g. LX2-32C, Yg-3-46a) exhibited their activity toward Pgp efflux pump and also β-III tubulin mediated drug resistance. However, they did not show greater specificity for the β-III tubulin isotype. Based on a C-seco taxane IDN5390 which was reported to be more selectively interacted with β-III tubulin than paclitaxel, we have succeeded in the design and synthesis of 7,9-O-linked C-seco taxanes with different linkages. Some of them exhibited better activity than IDN5390, and the activities were found sensitive to the structural changes of the linkers.

Citation Format: Yong Tang, Pei Cai, Javier Rodríguez-Salarisch, J. Fernando Díaz, Weishuo Fang. C-seco taxanes with 7, 9-linkages exhibit better activity in drug-resistant tumor cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3214. doi:10.1158/1538-7445.AM2014-3214
Abstract 3215: Role of pterostilbene on proliferation, invasion, and migration in ovarian and uterine cancer cells
Authors: Lowe Gina , Wen Wei , Roberts Cai , Finlay James , Glackin Carlotta , Han Ernest , Dellinger Thanh H.
Published: May 2015

PURPOSE: To determine the functional effects of the resveratrol analog, pterostilbene (PTE), and its potential adjuvant role with cisplatin, in ovarian and uterine cancer cells.

EXPERIMENTAL DESIGN: The human ovarian cancer cell lines (OVCAR8, SKOV3) and uterine cancer cell lines (HEC1A, ECC1) were obtained from ATCC, and grown according to standard protocols using recommended media. Standard proliferation (MTT) and migration assays were performed in routine fashion, with or without PTE +/- cisplatin treatment. For the in-vitro Cell Proliferation Assay, different concentrations of cisplatin (3 - 50 μM), pterostilbene (PTE) (30 - 500 μM), or both in combination were added for 72 hours. Cell migration was assayed using 24-mm diameter chambers with 8-μm pore filters, with or without treatment of PTE and cisplatin, and migrated cells were counted using Image-pro program. A synergistic analysis from the MTT assay results was calculated via the CompuSyn for Windows computer program based on the combination index (CI) theorem of Chou-Talalay, to determine the presence of synergy between cisplatin and PTE.

RESULTS: PTE reduces proliferation in OVCAR8, SKOV3, HEC1A, and ECC1 cells, and its anti-proliferative effect approaches that of cisplatin. Addition of PTE to cisplatin potentiates the anti-proliferative effects of cisplatin in HEC1A, ECC1, SKOV3, and OVCAR8 cells. The combined effect of PTE and cisplatin is synergistic in OVCAR8 cells based on synergy calculations (CI<1). PTE reduces cell migration in HEC1A and OVCAR8 cells, and its addition to cisplatin treatment improves the anti-migratory effect.

CONCLUSIONS: Pterostilbene, a resveratrol analog with improved bioavailability, reduces proliferation and migration in ovarian and uterine cancer cells, and potentiates the anti-proliferative and anti-migratory effects of cisplatin, thus presenting as an attractive non-toxic alternative for potential adjuvant chemotherapy in ovarian and uterine cancer.

Citation Format: Gina Lowe, Wei Wen, Cai Roberts, James Finlay, Carlotta Glackin, Ernest Han, Thanh H. Dellinger. Role of pterostilbene on proliferation, invasion, and migration in ovarian and uterine cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3215. doi:10.1158/1538-7445.AM2014-3215
Abstract 3221: Covalent inhibitors of S100B (SBiXs) in malignant melanoma
Authors: Cavalier Michael C.
Published: May 2015

Summary: The calcium binding protein, S100B, has been directly linked to malignant melanoma (MM) and has long been used as a prognostic indicator. Studies have proven S100B to not only be a clinical marker within MM, but that it binds to the tumor suppressor p53, and promotes its degradation. p53 is typically found as wild-type in MM, and inhibition of S100B expression restores both p53 protein levels and normal transcriptional activation/apoptosis activities. Subsequently, we have made it our long-term goal to discover S100B inhibitors (SBiXs) in order to restore the p53 tumor suppression function in patients with MM, and have already identified many SBiXs.

Purpose: It is our purpose to discover/synthesize and/or to improve existing inhibitors of S100B to restore p53 activity in human malignant melanoma. Amongst these SBiXs are compounds which covalently modify S100B. The effects of covalent SBiXs on S100B in Malignant Melanoma are to be investigated.

Results and conclusions: The development of S100B inhibitors (SBiXs) is ongoing at the University of Maryland Schools of Pharmacy and Medicine with the goal of improving affinity and specificity towards inhibiting S100B. Fluorescent Polarization Completion Assays have identified compounds that bind S100B and inhibit melanoma cell growth. S100B binding by these SBiXs was confirmed using NMR spectroscopy. The crystallographic structures of the SBiX:S100B complexes were solved revealing covalent modifiers occupying persistent binding site 2. The compounds offer potential bridging scaffolds between sites 1 and 3. All five SBiXs demonstrated the ability to kill malignant melanoma with some killing in a S100B dependent manner. TheS100B-SBiX complexes presented will act as the basis for the design of improved SBiXs. SBiXs may also have therapeutic value for treating other cancers with elevated S100B and wt p53 such as astrocytoma, renal tumors, and some forms of leukemia.

Citation Format: Michael C. Cavalier. Covalent inhibitors of S100B (SBiXs) in malignant melanoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3221. doi:10.1158/1538-7445.AM2014-3221
Abstract 3222: Structural evaluation of several antitumor Tencon variants
Authors: Cardoso Rosa , Goldberg Shalom , Luo Jinquan , Barnakov Alexander , Swift Edward , Jacobs Steven , Gilliland Gary
Published: May 2015

Tencons are engineered alternative scaffold proteins that combine the affinity and specificity of antibodies with the biophysical features of smaller molecules and hold high promise as biotherapeutic agents. We present the crystal structures and antigen binding data of several anti-Tumor Tencon variants. The structures of the variants show a switch of the FG loop conformation from α-helical at the tip to mostly β-strands. This conformational change prompts differential exposure of potential binding residues and a finger-like shape for the FG region that may facilitate its access to binding pockets on the target protein, thus likely accounting for the observed differences in KD.

Citation Format: Rosa Cardoso, Shalom Goldberg, Jinquan Luo, Alexander Barnakov, Edward Swift, Steven Jacobs, Gary Gilliland. Structural evaluation of several antitumor Tencon variants. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3222. doi:10.1158/1538-7445.AM2014-3222
Abstract 3227: JNK2 oligomerization regulates its activation through non-canonical pathways
Authors: Kaoud Tamer S. , Riggs Austin F. , Dalby Kevin N.
Published: May 2015

C-Jun N-terminal kinases (JNKs) regulate various cellular functions. Their activation is mediated cooperatively by both MKK4 and MKK7. The c-Jun N-terminal kinase 2 (JNK2) isoform is constitutively activated in glial tumor cell lines and human glioblastoma models. We investigated the regulation of JNK2 self-activation in vitro in both cell-free and cell-based experiments. Light scattering analysis suggested that unphosphorylated recombinant JNK2α2 exists in solution as a mixture of monomers, dimers and tetramers. JNK2α2 self-phosphorylates more rapidly in vitro when assayed at low concentration leading to its activation. However, at a slightly higher concentration, the formation of a tetramer is favored, whose ability to self-phosphorylate is suppressed. HEK293T cells that were transfected with varying amounts of pcDNA-JNK2α2 showed that an increased concentration of JNK2α2 (54 kDa) could suppress the appearance of activated JNK2α2 in mammalian cells. A mutant of JNK2 (F170R) thought to stabilize the DFG-out conformation appears to stabilize the formation of a JNK2 tetramer. Taken together our data suggest that oligomerization may regulate non-canonical (self-activation) activation of JNK2.

Citation Format: Tamer S. Kaoud, Austin F. Riggs, Kevin N. Dalby. JNK2 oligomerization regulates its activation through non-canonical pathways. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3227. doi:10.1158/1538-7445.AM2014-3227
Abstract 3230: Genome-wide localization of anti-cancer drugs
Authors: Anders Lars , Guenther Matthew G. , Qi Jun , Fan Zi Peng , Marineau Jason J. , Rahl Peter B. , Lovén Jakob , Sigova Alla A. , Smith William B. , Lee Tong Ihn , Bradner James E. , Young Richard A.
Published: May 2015

A vast number of small-molecule ligands, including therapeutic drugs under development and in clinical use, elicit their effects by binding specific proteins associated with the genome. An ability to map the direct interactions of a chemical entity with chromatin genome-wide could provide new and important insights into the mechanisms by which such small molecules interfere with tumor cell functions. We have developed a method that couples affinity capture of chemical entities and massively parallel DNA sequencing (Chem-seq) to identify the sites bound by small molecules throughout the human genome. Using Chem-seq, we have uncovered the full repertoire of the genomic sites bound by a BET bromodomain inhibitor, a cyclin-dependent kinase (CDK) inhibitor and a DNA intercalating drug. Moreover, by combining Chem-seq with ChIP-seq, we have characterized the interactions of drugs with their targets throughout the genome of tumor cells. These methods provide a powerful approach to enhance understanding of therapeutic action and characterize the specificity of drugs that interact with DNA or genome-associated proteins.

Citation Format: Lars Anders, Matthew G. Guenther, Jun Qi, Zi Peng Fan, Jason J. Marineau, Peter B. Rahl, Jakob Lovén, Alla A. Sigova, William B. Smith, Tong Ihn Lee, James E. Bradner, Richard A. Young. Genome-wide localization of anti-cancer drugs. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3230. doi:10.1158/1538-7445.AM2014-3230
Abstract 3234: Covalent capture of protein binding partners using an azide-tagged, photo-reactive stapled alpha helical p53 peptide
Authors: Whiting Amanda L. , Mitala Joe J. , Headley Kathryn M. , Reilly Joe , Morrison Bethanie L. , Murray Kevin A. , Bernal Federico
Published: May 2015

Mutations in the DNA binding region of tumor suppressor p53 render it transcriptionally inactive (dominant negative) but can also manifest in transcriptionally-independent gain-of-function (GOF) effects, possibly due to mutp53 interacting with other, as yet, unknown targets. Identification of these targets would help to elucidate the pathways and vulnerabilities involved in mutp53 GOF. Covalently linking a known handle to these unknown proteins is one method to aid in their isolation and identification. A hydrocarbon-stapled alpha helical peptide of the p53 transactivation domain (residues 14-29), SAH-p53-8, has been shown to strongly interact with known p53 targets that utilize this domain for binding (HDM2, Kd = 55 nM, HDMX, Kd = 2.3 nM). Substitution of a key interacting residue for the unnatural amino acid benzoylphenylalanine (Bpa) results in a stapled peptide that retains the biochemical properties of SAH-p53-8 (affinities of 38.1 nM and 58.3 nM, respectively) and can covalently capture known targets via photochemical reaction. The introduction of an azido group - a versatile, biochemically-inert capping group - to the N-terminus of this photo-reactive peptide allows for the attachment of various tags to the covalently bound protein(s) via a copper-catazlyzed azide alkyne cycloaddition. Tags include both fluorophores for visualization and affinity tags for protein isolation. This poster will cover the biochemical proof-of-principle of this protein isolation method and exploration into more complex cell lysate environments.

Citation Format: Amanda L. Whiting, Joe J. Mitala, Kathryn M. Headley, Joe Reilly, Bethanie L. Morrison, Kevin A. Murray, Federico Bernal. Covalent capture of protein binding partners using an azide-tagged, photo-reactive stapled alpha helical p53 peptide. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3234. doi:10.1158/1538-7445.AM2014-3234