bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5530: Fluorescence laparoscopy improves metastatic detection in both early and advanced colon cancer
Authors: Menen Rhiana , Kaushal Sharmeela , Snyder Cynthia , Hoffman Robert M. , Bouvet Michael
Published: August 2012

The aim of the present study was to improve detection and treatment of colon cancer with the use of fluorescence laparoscopy in an orthotopic mouse model of human colon cancer. The orthotopic model was established by intra-cecal injection of GFP-expressing HCT-116 human colon cancer cells into 12 six-week-old female athymic mice. Two models were then established: one of early disease and the second of widely metastatic disease. Two weeks after implantation, 6 mice underwent diagnostic laparoscopy. Laparoscopy was performed first under standard brightfield lighting, followed by fluorescent lighting. The number of metastatic foci identified within each of the four quadrants of the peritoneal cavity was recorded in three mice in each light mode. This process was repeated in the second group 4 weeks after implantation. Following laparoscopy, all animals were sacrificed. Fluorescence imaging was used as a positive control to identify metastasis at necropsy. Tumors and metastases were collected and processed for histologic review. Fluorescence laparoscopy enabled visualization of colon cancer metastatic foci which could not be visualized with standard brightfield laparoscopy in both the early (p= 0.028) and late (p= 0.001) models of colon cancer. When compared to the positive control of open fluorescence imaging, standard brightfield laparoscopy was significantly inferior in both the early (p= 0.03) and late (p= 0.0005) groups. Conversely, fluorescence laparoscopy was not significantly different from open fluorescence imaging in either the early (p= 0.94) or late (p= 0.56) groups. Fluorescence laparoscopy thus allowed identification and localization of sub-millimeter micrometastases that could not be distinguished from surrounding tissue under brightfield laparoscopy. Metastatic lesions were histologically confirmed. This report illustrates the important clinical potential of fluorescence laparoscopy in the surgical treatment of cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5530. doi:1538-7445.AM2012-5530
Abstract 5537: Quantitative multiplexed SRM analysis of oncogenic receptors in FFPE colorectal carcinoma tissue
Authors: Hembrough Todd , Thyparambil Sheeno , Liao Wei-Li , Darfler Marlene , Krizman David , Adams Richard , Burrows Jon
Published: August 2012

Multiple receptor tyrosine kinases are the target of either approved drugs (Her2, EGFR) or robust clinical development efforts (cMet, IGF1R, Her3, etc). In many cases initial drug response is followed by resistance and tumor progression. There are many mechanisms which have been proposed for this including co-expression of alternate RTKs that continue to stimulate tumor growth in the presence of EGFR/Her2 inhibition. Because of the potential that co-expression of oncogenic receptors mediates resistance in these clinical trials, we have developed a panel of new SRM assays which measure the expression of these critical RTKs in multiplex from FFPE tissues. These assays are based on the Liquid Tissue®-SRM technology platform. This approach enables relative and absolute quantification of proteins and their phosphorylation status directly in formalin fixed paraffin embedded (FFPE) tissue. Here we describe the quantitative multiplexed analysis of EGFR, IGF1R and cMet expression in a cohort of 75 archival FFPE colorectal cancer tumors from Cetuximab treated patients. This analysis identified 7 different fingerprints of expression of these three targets. Since each of these targets has either approved inhibitors, or highly active clinical development programs, we are hopeful that this analytical approach can help to identify patients who are most likely to respond to single or combination therapies of RTK inhibitors Follow up studies are underway to expand the RTK- multiplex, repeating EGFR, IGF1R and cMet and adding Her2, Her3, Her4, DR5, cSrc, and other drug targets into the multiplex. Our intention is to use this broader multiplex to re-analyze these patients and attempt to correlate the multiplexed expression of these targets with drug response data.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5537. doi:1538-7445.AM2012-5537
Abstract 5542: Gene expression and genomic DNA stabilization in whole blood stored at room temperature
Authors: Liberal Vasco , Wilkinson Steven P. , Stassinopoulos Angela , Huang Winnie , Whitney Scott , Shireen Lutfunnessa , Ohgi Sharron , Muller Rolf , Muller-Cohn Judy
Published: August 2012

Genomic and mitochondrial DNA and gene expression profiles from blood samples are increasingly being used to diagnose specific diseases, including cancer, monitor disease progression and assess patient responses to medical treatment. However, such applications require reliable preservation of total RNA and DNA in patient samples during collection, transport and storage. Numerous agents threaten DNA and RNA profiles in collected blood samples, including nucleolytic attack, oxidative damage and hydrolysis. Such damage can severely disrupt genotyping and DNA sequencing efforts. Transcription profiles are highly dynamic and can change rapidly during and after blood collection using current methods, potentially affecting interpretation of the expression analysis, and ultimately lead to inadequate treatment options. In this study, we compare freezer storage with room temperature stabilization formulations, DNAgard Blood and PAXgene Blood DNA, in their capacities to preserve genomic DNA integrity in human whole blood for 8 months. In a separate analysis, we compare freezer storage with room temperature stabilization formulations, PAXgene Blood RNA and Biomatrica formulation DRG, in their capacities to stabilize RNA in whole blood samples. We assess changes in the relative expression of a panel of 34000 genes in whole blood specimens over the course of 7 days using RT-qPCR and the Human HT-12 Bead Array (Illumina). Our results demonstrate that room temperature blood storage is a valid alternative to cold-storage for preserving gDNA for more than a year, and RNA for at least 7 days.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5542. doi:1538-7445.AM2012-5542
Abstract 5689: Anti-E2F-1 PEGylated liposomal encapsulated peptide regresses human small cell lung cancer & lymphoma xenografts
Authors: Xie Xiaoqi , Banerjee Debabrata , Minko Tamara , Johnson-Farley Nadine , Kerrigan John E. , Ercikan-Abali Emine , Scotto Kathleen W. , Bertino Joseph R.
Published: August 2012

The E2F family of transcription factors is critical to large number of key cellular processes, including replication, DNA repair, and differentiation. E2F-1 is unique among the E2F family of transcription factors in that it can act as a tumor suppressor or as an oncogene, depending on the cellular context. Given the critical functions of E2F target genes in replication, and differentiation, it is perhaps not surprising that dysregulation of E2Fs has been associated with the cancer phenotype. E2F-1,2 and-3, “activating” E2Fs, function as heterodimers with DP proteins and are regulated by the retinoblastoma protein. E2F-1 and 3 are over expressed in many human tumors and overexpression (suggesting oncogene addiction) is associated with a poor prognosis. We have used phage display to obtain peptides that bind tightly to the E2F consensus promoter recognition sequence. When one of these peptides was coupled to a modified penetration sequence to enhance uptake, the penetratin- peptide (PEP) potently inhibited some but not all malignant cell lines. Modeling studies and ChIP studies showed that part of the pentratin sequence was involved in the binding of the PEP to the E2 F1 promoter. The PEP down-regulated E2F-2 and 4, as well as E2F-1, and-down stream targets of E2F-1. The small cell lung carcinoma (SCLC) cell line and the Ramos cell line (Burkitt lymphoma) were sensitive to low micromolar concentrations of the PEP, and apoptosis was seen within 6 h. These cell lines have mutant p53, and over express E2F-1 and cMyc and BCL2, and are “oncogene addicted”. Normal cells (human bone marrow CD34+ hematopoeitic stem cells, human marrow mesenchymal cells and MEFs), were insensitive to high concentrations of the PEP. When encapsulated in PEGylated liposomes, the PEP caused marked regression of advanced SCLC xenografts in mice without appreciable toxicity. These results prompt us to explore further preclinical development of the PEP for tumor therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5689. doi:1538-7445.AM2012-5689
Abstract 5710: Effective siRNA delivery through specific targeting of tumor
Authors: Jun Eun Sung , Kim Jong-Ho , Lee Byung-Heon , Park Rang-Woon , Kim In-San
Published: August 2012

The siRNA has been expected to apply for several diseases such as cancer since siRNA specifically silences the disease-associated genes. However, effective gene carriers should be developed to overcome the low siRNA stability in vivo, form stable complexes and facilitate intracellular uptake of siRNA. Here, we have investigated tumor targetable oligopeptide carrier including tumor-homing peptide, AP-1, and rich positive peptide, nine arginines, for systemic siRNA delivery. Phage display technique is a promising tool for selecting peptides or proteins with specific binding properties from a lot of variants. AP-1 peptide can bind with interleukin-4 receptor which expressed on surface of several human cancer cells like MDA-MB231. Arginine rich peptides have been widely used for forming complex with DNA, which promote efficient transfection in various mammalian cells. Oligopeptides formed ionic complex with siRNA at 150 nm size and these peptide/siRNA complexes were transfected in MDA-MB231. Under in vitro silencing experiments, peptide/siRNA complexes did silencing of Dronpa gene as similar to lipofectamine/siRNA complexes, which were observed by fluorescent microscope, flow cytometry, and western blotting analysis. For in vivo experiments, we prepared tumorbearing mice by inoculation of Dronpa-MDA-MB231 cells (Dronpa-stable cells). Tumor targetable oligopeptide/siRNA complexes were administrated through tale vein when tumor volume reached at 50 mm3 - we observed fluorescent signals of Dronpa in tumor. In tumor targetable oligopeptide/siRNA-treated mice, weak signals of Dronpa were observed compared with free siRNA-treated mice. These results revealed the promising potential of our oligopeptide carrier as a stable and effective siRNA delivery system for cancer treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5710. doi:1538-7445.AM2012-5710
Abstract 5717: The Poly (ADP-Ribose) Polymerase-1 Inhibitor, AZD2281, enhances radiation response of breast cancer cells by inhibiting DNA repair
Authors: Mehta Meghna , Munshi Anupama
Published: August 2012

PARP-1 is a nuclear enzyme essential for base excision repair (BER), SSB, and possibly other DNA repair processes and an important damage sensor. It has been identified as a synthetic lethal target in BRCA1/2 breast cancers. PARP-1 is activated after binding DNA strand breaks and uses NAD+ as a substrate to form long branched polymers of poly(ADP-ribose) (PAR). PARP-1 mediated poly(ADP-ribosylation) recruits various nuclear acceptor proteins, such as XRCC1, histones, and PARP-1 itself, to assemble other repair complexes to execute DNA repair. As a result, PARP inhibitors are being developed for cancer therapy due to their ability to sensitize cells to DNA-damaging agents. Thus, in this current study, we hypothesized that enhanced radiosensitivity of breast cancer may be achieved by combining ionizing radiation (IR) with the PARP inhibitor, AZD2281. We studied the effects of combined treatment of radiation and AZD2281 on three human breast cancer cell lines, MCF-7, MDA-MB-231 and Hs578t. Cells were treated either with vehicle alone or with AZD2281 (4μM for 24hrs for MCF-7; 2μM for 24hrs for the MDA-MB-231 and Hs578t cells). Clonogenic cell survival assay showed that AZD2281 enhanced tumor cell radiosensitivity with the survival factor at 2Gy (SF2) being reduced from 26%, 52.8%, and 51.5% in vehicle treated to 18.5%, 27.8%, and 45.52% in AZD2281-treated MCF-7, MDA-MB-231 and Hs578t cells respectively. As an initial investigation into the mechanism responsible for AZD2281-mediated radiosensitization, we examined the effect of AZD2281 treatment on the expression of proteins known to be involved in the repair of radiation-induced DSBs. Western blot analysis for Ku70, Ku86, and DNA-PKcs demonstrated a decrease in the levels of all three proteins following AZD2281 treatment in the breast cancer cell lines tested. As an additional test of our hypothesis that AZD2281 impairs the repair of damaged DNA, γ-H2AX foci were assessed as indicators of DNA damage. PARP inhibition markedly enhanced γ-H2AX persistence in the MCF-7 and MDA-MB-231 cell lines suggestive of the presence of persistent DSBs. These data support the use of the PARP inhibitor, AZD2281, in combination with radiation therapy as an innovative treatment strategy to potentially improve outcomes in breast cancer patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5717. doi:1538-7445.AM2012-5717
Abstract 5728: The synergistic effect of Cucurbitacin B and radiation treatment
Authors: Zhang Mei , Yin Liangjie , Yang Shanmin , Hong Jinsheng , Chen Chun , Han Deping , Hou Yanqiang , Zhang Bingrong , Huang Luqiang , Zhang Amy , Vidyasagar Sadasivan , Okunieff Paul , Zhang Lurong
Published: August 2012

Cucurbitacin B (CB), a small molecule purified from Trichosanthes kirilowii Maximowicz, possesses a potent antitumor effect. In this study, we investigated the combined effect of CB and ionizing radiation (IR) on aggressive 4T1 breast cancer cells. In a clonogenic assay, different doses of CB (0, 1.7, or 5 nM) and IR (0, 2, 4, or 6 Gy) were applied to 4T1 cells either in a single regimen or in combination. The surviving fraction at 2 Gy was greatly reduced with the increased doses of CB. The results calculated with CompuSyn program indicated that the combination index (CI) was <1 and the dose reducing index (DRI) was >1, suggesting that the combination of CB and IR has a synergistic effect against cancer cells. Our studies of the underlying mechanism revealed that: 1) CB enhanced IR-induced loss of mitochondria potential, as evidenced by a 10-20% increase in the transition of aggregative JC-1 (representing healthy cells) into monomeric JC-1 (representing early apoptotic cells); 2) cell-cycle analysis revealed that although samples treated with CB or 3 Gy alone had 38% or 29% of cells, respectively, in the G2-M phase as compared to 23% of the vehicle control, the combined treatment encouraged 51% of cells to accumulate in the G2-M phase; and 3) the Western blot assay indicated that survival-related molecules (e.g., phosphorylated Stat 3, c-Myc, Bcl-2, and Bcl-xL) were downregulated, while apoptosis-related molecules (e.g., caspase-9, p21, and p53) were upregulated in the combined treatment group. The data reveal that the systemic administration of CB will not only stop the spread of micrometastases but also enhance local tumor control, which will result in better treatment outcomes. The clinical value of CB should be further explored.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5728. doi:1538-7445.AM2012-5728
Abstract 574: α-santalol induces caspase-dependent apoptosis and G2/M phase cell cycle arrest in human breast cancer cells
Authors: Santha Sreevidya , Bommareddy Ajay , Rule Brittny , Guillermo Ruth , Kaushik Radhey S. , Young Alan , Dwivedi Chandradhar
Published: August 2012

Alpha-santalol, an active component of sandalwood oil, has been shown to have chemopreventive effects on skin cancer both in vitro and in vivo. However, effects of alpha-santalol on other types of cancer have not been studied. In this study, we examine the effects and mechanisms of action of alpha-santalol on human breast cancer cells using estrogen receptor (ER)- positive (MCF-7) and estrogen receptor- negative (MDA-MB-231) breast cancer cell lines. MTT and BrdU cell proliferation ELISA results showed inhibition of cell viability and proliferation in a time and dose-dependent manner in both cell lines. Alpha-santalol at 25-150 µM concentration decreased cell viability after 12h treatment. In addition, TUNEL assay and flow cytometry results revealed that alpha-santalol significantly induced apoptosis in MCF-7 and MDA-MB-231 cells. Further, immunoblotting and caspase activity assays showed involvement of caspase-3, caspase-8 and caspase-9 in apoptotic cell death. Induction of apoptosis by alpha-santalol was further confirmed by detecting the cleavage of Poly (ADP-ribose) Polymerase (PARP) through western blotting. Cell cycle distribution of alpha-santalol treated MFC-7 and MDA-MB-231 cells were analyzed by fluorescence- activated cell sorting (FACS) analysis of propidium iodide staining. We observed that alpha-santalol treatment arrests cell cycle at G2/M phase at 25µM −75µM concentration in both cell lines. Taken together, our studies reveal a potential mechanism for the chemopreventive effect of alpha-santalol on ER- positive and ER- negative breast cancer cells through induction of apoptosis and cell cycle arrest at G2/M phase. Alpha-santalol could be effective for the prevention and treatment of breast cancer. (Supported by Translational Cancer Research Center funded by South Dakota, Governors Office of Economic Development)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 574. doi:1538-7445.AM2012-574
Abstract 5743: Positron emission tomography (PET) imaging analogs of glutamine: Development and evaluation as probes for cancer imaging
Authors: Buck Jason R. , Saleh Sam , Hight Matthew R. , Nickels Mike L. , Manning H. Charles
Published: August 2012

Emerging evidence suggests that nutrient uptake in cancer cells is controlled by oncogenic signaling pathways. The natural amino acid glutamine is an essential nutrient for cell growth and proliferation. Glutamine also participates in protein synthesis and is a nitrogen source for the production of select nucleotides and amino acids. Consequently, quantitative measures of glutamine uptake may reflect critical processes in oncology. Recently, a fluorinated glutamine analog for positron emission tomography (PET) was shown to possess both highly specific cellular uptake in glioma cell lines and in vivo tumor localization (J Am Chem Soc 2011;133:1122-1133). Synthetic methodology leading to this agent, an optically pure 4-fluoro-glutamine derivative, was also reported. Though effective, this convergent synthesis requires multiple steps, extensive chromatographic separation, and nearly a week of reaction time. Employing a chiral auxiliary Schiff base and microwave-assisted organic synthesis, we have significantly improved the synthesis of this and similar agents, drastically reducing the number of synthetic steps (from ten to four) and the overall reaction time (from seven days to two hours). While the radiochemistry relies upon traditional box methods, we are also currently employing high-throughput microfluidic radiolabeling strategies to expedite probe production and labeling efficiency. Finally, using optically pure [18F]4-FGln (2S,4R) synthesized by our improved synthesis, we report preclinical PET images of models of human colorectal cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5743. doi:1538-7445.AM2012-5743
Abstract 577: Equol enhances tamoxifen's antitumor effect by induction of caspase-mediated apoptosis in MCF-7 breast cancer cells
Authors: Charalambous Christiana , Constantinou Andreas I.
Published: August 2012

Soy phytoestrogens, such as daidzein and its metabolite equol, have been proposed to be responsible for the low breast cancer rate in Asian women. However, since these compounds possess both estrogenic and anti-estrogenic properties, it remains unknown whether they provide protection against tumor development and progression. Since the majority of estrogen receptor positive (ER+) breast cancer patients are treated with the selective estrogen receptor modulator tamoxifen, the basic objective of this project is to determine whether equol enhances or inhibits tamoxifen's anti-tumor effect and to identify the molecular mechanisms involved. For this purpose, we examined the individual and combined effects of equol and tamoxifen on the ER+ breast cancer cell line MCF-7 (ERα+/−Rα+). We found that equol at concentrations higher than 10 μM significantly reduced the viability of MCF-7 cells. Furthermore, the combination of equol (100 μM) and 4-hydroxytamoxifen (4-OHT; 10 μM) reduced MCF-7 cell viability and induced apoptosis more effectively than each compound alone. Subsequent treatment of MCF-7 cells with the pan-caspase inhibitor Z-VAD-FMK and the calpain/cathepsin inhibitor ALLN demonstrated that equol and 4-OHT induce apoptosis via both caspase-depended and -independed mechanisms. In order to further investigate the apoptotic pathways employed by equol, 4-OHT and their combination, we evaluated the expression of key proteins involved in apoptosis such as PARP, α-fodrin, AIF, bcl-2, bax, p21 and p53 using Western blot analysis. The combination of 4-OHT and equol induced PARP and α-fodrin cleavage which was inhibited by Z-VAD-FMK indicating that their combination induces caspase-depended apoptosis. This was accompanied by reduced bcl-2:bax ratio confirming a pro-apoptotic effect. The combination of equol and 4-OHT induced caspase-7 cleavage indicating a significant role for caspase-7 in equol- and tamoxifen-induced apoptosis. Finally, upon determination of the effect of equol and tamoxifen on estrogen receptor expression in MCF-7 cells, it was found that equol reduces the ERα:ERα ratio, while 4-OHT increases it. When cells were treated with both compounds we observed that the effect of equol prevailed, providing an additional explanation for the beneficial combined effects of equol and tamoxifen. In conclusion, the effects of equol in potentiating tamoxifen's anti-tumor effect may be explained by the induction of the caspase-mediated apoptotic pathway and the reduction of the ERα:ERα ratio. These results can find applications in clinical or pre-clinical cancer chemoprevention studies against breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 577. doi:1538-7445.AM2012-577
Abstract 586: Withaferin-A inhibits prostate tumor growth by enhancing anti-tumor immune response
Authors: Singh Rajesh , Grizzle William E. , Lillard James W. , Singh Shailesh
Published: August 2012

Indolent nature of prostate cancer provides a ‘large window’ of opportunity for chemopreventive strategies. Most chemopreventive strategies are primarily focused on controlling and/or reversing the mechanisms involved in neoplastic transformation and cancer progression. It is now well accepted that tumors evade the patient's defense system during progression and growth at distant sites. Mounting evidences suggest immunological defects in prostate cancer patients with higher numbers of functionally suppressive CD4+CD25+FOXP3+ T cells (Treg) and lower number of effectors T cells. Furthermore, tumor microenvironment exerts a profound immunosuppressive effect on effectors T cells by secreting TGF-β and IL-10. Therefore, modulating TGF-β and IL-10 using natural and dietary components constitutes an innovative approach for prostate cancer prevention and treatment. In this study, we have shown that Withaferin-A (WA), a bioactive compound isolated from Withania Somnifera also called “Ashwagandha” has the ability to modulate immunosuppressive cytokines (IL-10 and TGF-β) produced by the prostate cancer cells. Additionally, we show selective inhibition of Treg proliferation and induction of apoptosis in Tregs cells in immune competent prostate cancer mouse model treated with WA compared to controls. Our results suggest that WA have potential to fine-tune immunosuppressive phenotype of tumor-associated lymphocytes by modulating immuno-suppressive cytokines. Significant inhibition of tumor growth in immune competent prostate cancer mouse model as compared to immune deficient xenograft suggest potential role of WA on reversal of anti-tumor immune response from immune suppression to immune competence, which is required to fight against prostate cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 586. doi:1538-7445.AM2012-586
Abstract 587: Phenethyl isothiocyanate suppresses inhibitor of apoptosis family protein expression in prostate cancer cells in culture and in vivo
Authors: Sakao Kozue , Desineni Sudhakar , Hahm Eun-Ryeong , Singh Shivendra V.
Published: August 2012

Bioactive food components continue to gain traction for possible chemoprevention of prostate cancer, which is a leading cause of cancer-related death among American men. Phenethyl isothiocyanate (PEITC) is one such compound that is under active investigation for chemoprevention of prostate cancer as well as other types of malignancies, including colon, lung, and breast cancers. PEITC occurs naturally as a thioglucoside conjugate (gluconasturtiin) in a variety of edible cruciferous vegetables (e.g., watercress), and is generated upon cutting or chewing of cruciferous vegetables due to myrosinase-catalyzed breakdown of the gluconasturtiin. Initial evidence supporting prostate cancer protective effect of cruciferous vegetables emerged from population-based case control studies. In vivo chemopreventive efficacy of PEITC against prostate cancer has now been demonstrated in a transgenic mouse model (Transgenic Adenocarcinoma of Mouse Prostate mice; hereafter abbreviated as TRAMP mice). Post-initiation cancer chemopreventive response to PEITC is associated with apoptosis induction but the mechanism of cell death is not fully understood. The present study was undertaken to determine the role of inhibitor of apoptosis (IAP) family proteins in regulation of PEITC-induced apoptosis. The IAP family proteins play critical roles in apoptosis regulation by inhibiting caspases. Elevated expression of IAP proteins, including X-linked inhibitor of apoptosis (XIAP) and Survivin has been reported in human prostate cancers. Exposure of PC-3 and LNCaP human prostate cancer cells to PEITC resulted in downregulation of XIAP and Survivin proteins and suppression of Survivin mRNA expression. PEITC administration to Transgenic Adenocarcinoma of Mouse Prostate mice caused modest but significant downregulation of XIAP and Survivin proteins in the dorsolateral prostate. Proapoptotic response to PEITC was significantly attenuated by ectopic expression of XIAP and Survivin proteins. In addition, Survivin overexpression conferred modest but significant protection against PEITC-mediated inhibition of PC-3 cell migration. In conclusion, these results indicate that cellular responses to PEITC, including apoptosis induction and inhibition of cell migration, in prostate cancer cells are mediated by downregulation of XIAP and/or Survivin, which may serve as valid biomarkers to assess PEITC response in future clinical investigations. This investigation was supported by the USPHS grant RO1 CA101753-08.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 587. doi:1538-7445.AM2012-587
Abstract 588: Evidence for a role of a P2X7- EHBP1-Akt axis in prostate carcinogenesis and in statin chemo-prevention
Authors: Ghalali Aram , Wiklund Fredrik , Darabi Hatef , Högberg Johan , Stenius Ulla
Published: August 2012

Prostate cancer (PCa) is the most commonly diagnosed cancer (over 30% of all cancer in men) and the second leading cause of cancer death in men. Statins, HMG-CoA reductase inhibitors, are potent cholesterol lowering drugs. Recent epidemiology suggests that statins prevent aggressive PCa development. In previous cell experiments we have shown that statins can down-regulate nuclear Akt responses to insulin (J Biol Chem 2010; 285, 27900). We showed that atorvastatin decreases nuclear phosphorylated Akt (pAkt) levels in pancreatic and lung cancer cell lines within minutes. These effects were mediated by the P2X7 receptor, and its natural ligand, extracellular ATP, induced the same effects as statins. Our published work also implicates PTEN, PHLPP1 and 2 and other phosphatases, which form protein complexes. Akt has oncogene activity in PCa and many other cancer types, and we have now tested the possibility that the P2X7-Akt pathway is of importance for the anticipated anticancer effects of statins in the prostate. We performed SNP analysis of two cohorts of PCa patients (Cgems and Gasp). We found significant associations for SNPs in Akt1, Caveolin1, P2X7, NEDD4-1 and EHBP1 genes. NEDD4-1 negatively regulates PTEN. EHBP1 is a previously shown risk factor for aggressive PCa, and has been implicated in plasma membrane trafficking. A signaling pathway analysis (Hum Hered 2011; 71, 11) also indicated significant associations to PCa (p = 0.043). EHBP1 showed the strongest influence. In additional cell studies we employed PCa cells and found that knock down of EHBP1 or p110α (previously shown to have oncogenic activity in PCa; Nature 2008; 454, 776) prevented depletion of nuclear Akt. In further long term cell experiments (several weeks) we found that high concentrations of statins resulted in a decreased expression of p110α. We also overexpressed a mutated NEDD4-1 and this treatment inhibited the effect of statin on nuclear pAkt in PCa cells (DU145). Taken together this study provides evidence for a role of a putative P2X7-EHBP1-Akt axis in PCa development. As this pathway is negatively regulated by statins it may have a role in the indicated chemo-preventive effect of statins.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 588. doi:1538-7445.AM2012-588
Abstract 590: Andrographolide suppresses prostate cancer cell migration and alters the expression of vimentin, ZO-1 and MMP-11
Authors: Banerjee Aditi , Casillas Joseph , Sanchez Maria , Rohena Krizia , Martinez-Ferrer Magaly
Published: August 2012

Prostate cancer is the most common cancer and the second cause of cancer-related deaths in males in the United States. The development of novel chemopreventive strategies may be extremely helpful to prevent and arrest the development of this neoplasm. Andrographolide, a labdane diterpenoid that is the main bioactive component of the medicinal plant Andrographis paniculata, has anti-inflammatory and anticarcinogenic properties. In this study, we sought to examine the effect of andrographolide on PC3 and LNCaP prostate cancer epithelial cells. Andrographolide significantly inhibited PC3 cell growth at a concentration of 10µM after 24 h of treatment (P < 0.001), and inhibited LNCaP cell growth at a concentration of 15 µM after 24 h of treatment (P < 0.001). Wound healing assay and boyden chamber experiments were performed in order to determine whether andrographolide had an effect on migration and invasion. When PC3 and LNCaP cells were treated with andrographolide the total number of migrating and invading cells was significantly reduced (P < 0.001) when compared to control. To study possible anti-migration and anti-invasion mechanisms of andrographolide on PC3 and LNCap cells, we examined the expression of various adhesion molecules. Andrograholide significantly decreased the expression of vimentin in PC3 and LNCap treated cells. However, PC3 and LNCaP cells treated with andrographolide had increased expression levels of occludin and ZO-1. In addition, immunofluorescence and western blot analysis showed that PC3 and LNCaP cells treated with andrographolide had decresased expression levels of MMP-11. Our results suggest that the anti-migration and anti-invasion effects of andrographolide in PC3 and LNCaP cells may be associated with alterations in the expression of occluding, ZO-1 and MMP-11.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 590. doi:1538-7445.AM2012-590
Abstract 593: Mechanisms for inhibition of rodent mammary carcinogenesis by wheel running and/or dietary energy restriction
Authors: Jiang Weiqin , Zhu Zongjian , McGinley John N. , Neil Elizabeth , Thompson Henry J.
Published: August 2012

Our laboratory has reported the inhibitory effects of combined physical activity (PA) and dietary energy restriction (DER) on mammary carcinogenesis while the associated mechanisms are rarely investigated. The objective of this experiment was to determine the cellular regulation of mammary cancer by PA and/or DER. Female Sprague Dawley rats were injected with 1-methyl-1-nitrosourea (50 mg/kg) and 7 days thereafter randomized to: 1) sedentary control, 2) PA at wheel running high (WR-HIGH, 3.4 km per day); or the energy balance equivalents of PA: 3) PA at wheel running low (WR-LOW, 1500 revolutions per day + DER), or 4) DER. In order to investigate the effects of physical activity at a constant intensity, rats were given free access to a motorized activity wheel (55 m/min) with reinforcement of running behavior using a food reward system. The duration of this experiment was 8 weeks. Overall, PA and/or DER reduced mammary cancer incidence, cancer multiplicity, and tumor burden and prolonged cancer latency (p<0.05) in comparison to the sedentary control group. PA, DER, or their combination differentially affected host systemic factors including bioavailable IGF-1 and adipokines (leptin and adiponectin) and cell autonomous mechanisms including modulation of FoxO and down regulation of Akt/mTOR signaling. Mediation appears to be via inhibition of cell proliferation and induction of apoptosis. Supported by PHS grant CA100693 from the National Cancer Institute.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 593. doi:1538-7445.AM2012-593
Abstract 603: Targeted Casp8AP2 methylation increased drug resistance in mesenchymal stem cell and cancer cells
Authors: Leu Yu-Wei , Pai Mei-Yu , Hsu Chia-Chen , Lee Kuan-Der , Chen Chih-Cheng , Chen Yao-Li , Huang Tim H.-M. , Hsiao Shu-Huei
Published: August 2012

Casp8AP2 is a FLASH protein that is critical for the formation of death complex and the relay of death signal into the cells. Genetic defect of Casp8AP2 was found in diseases like acute lymphoblastic leukemia, implying that the miss-expression of Casp8AP2 might cause disease. On the other hand, the promoter of Casp8AP2 is enriched with CpG islands and we observed differential methylation changes within Casp8AP2 promoter during mesenchymal stem cell (MSCs) differentiation. Aberrant DNA methylation was also identified within the Casp8AP2 promoter in vivo like in liver cancers. It was then hypothesized that the abnormal DNA methylation within the Casp8AP2 promoter might contribute to the enhanced survival or drug resistance in cancer development. We determined the epigenetic states within the Casp8AP2 promoter in different cells and confirmed that there are methylation differences between the normal and tumoral samples within the Casp8AP2 promoter. Differential methylation was also observed between the drug resistant cancer cells and primary cancer cells. Global demethylation in drug resistant cell lines reversed the Casp8AP2 methylation, which was correlated with the reversed resistance phenotype. We then target-methylated the Casp8AP2 promoter in normal somatic stem cells and cancer cells like MDA-MB-231 and found increased drug resistance in these cells. These data point out that methylation within the death complex like Casp8AP2 might be able to serve as biomarker and treatment target for drug resistance in cancer cells. (Supported in part by NSC100-2320-B-194-001)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 603. doi:1538-7445.AM2012-603
Abstract 61: A new serotype chimera Cancer Terminator Virus (Ad.5/3-CTV) expands the efficiency and specificity of prostate cancer gene transfer and therapy
Authors: Azab Belal , Dash Rupesh , Das Swadesh , Bhutia Sujit , Shen Xue-Ning , Quinn Bridget , Dasgupta Santanu , Dent Paul , Dmitriev Igor , Wang Xiang-Yang , Curiel David , Grant Steven , Pellecchia Maurizio , Reed John , Sarkar Devanand , Fisher Paul
Published: August 2012

Conditionally replication-competent adenoviruses (CRCAs) represent potentially useful reagents for treating prostate cancer (PC). We previously constructed a unique CRCA, Cancer Terminator Virus (CTV), in which the E1A gene, necessary for adenoviral replication, is controlled by the cancer-selective Progression Elevated Gene-3 (PEG-3) promoter and which simultaneously expresses melanoma differentiation associated gene-7/interleukin-24 (mda-7/IL-24) from the E3 region of the adenovirus. mda-7/IL-24, a novel member of the IL-10-related cytokine gene family, displays almost ubiquitous antitumor properties in vitro and in vivo, leading to its rapid entry into the clinic, where its safety and clinical efficacy, when administered by adenovirus (Ad.mda-7; INGN 241), was observed in a phase I clinical trial in humans with advanced carcinomas and melanomas. mda-7/IL-24 preferentially induces apoptosis in cancer cells while exerting no discernible toxic effects toward normal cells and it also elicits potent “antitumor bystander activity” in distant cancer cells as a consequence of autocrine and paracrine secretion of MDA-7/IL-24. The CTV was generated on a serotype 5 background (Ad.5-CTV) with infectivity depending on Coxsackie-Adenovirus Receptors (CARs). CARs are frequently reduced in many tumor types, including PCs thereby limiting effective therapy. Using serotype chimerism, a novel CTV (Ad.5/3-CTV) was created by replacing the Ad.5 fiber knob with the Ad.3 fiber knob facilitating infection in a CAR-independent manner. We evaluated Ad.5/3-CTV in comparison with Ad.5-CTV in low CAR human PC cells, demonstrating higher efficiency in inhibiting cell viability in vitro. Moreover, Ad.5/3-CTV potently suppressed in vivo tumor growth in a nude mouse xenograft model and in a spontaneously induced PC in Hi-myc transgenic mice. Ad.5/3.CTV exerted a marked anti-tumor ‘bystander’ effect in vivo confirming therapeutic utility. In these contexts, and considering the significant responses in a Phase I clinical trial of a non-replicating Ad.5-mda-7, the use of Ad.5/3-CTV to administer therapeutic and cytotoxic mda-7/IL24 to selectively induce cytolysis and apoptosis in prostate tumor cells represents a viable treatment options. (Supported by R01 CA097318 and P01 CA104177)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 61. doi:1538-7445.AM2012-61
Abstract 619: Flavokawain A inhibits urinary bladder carcinogenesis in the UPII-SV40T transgenic mouse bladder cancer model.
Authors: Liu Zhongbo , Li Xuesen , liu Shuman , Xu Xia , Tian Xuejiao , Simoneau Anne R. , Wu Xue-Ru , Zi Xiaolin
Published: August 2012

The UPII-SV40T transgenic model of the mouse bladder reproduces a precursor to recurrent and invasive Transitional Cell Carcinoma (TCC): carcinoma in situ (CIS). This model therefore is useful for testing the potential of agents for preventing bladder cancer recurrence. Flavokawain A is the predominant chalcone identified from the kava plant. We have recently shown that flavokawain A exhibited strong growth inhibitory effects on human bladder cancer cell lines both in vitro in cell cultures and in vivo in a xenograft mouse model. In this study, we examine whether flavokawain A can also inhibit bladder cancer development in the UPII-SV40T transgenic model. The transgenic mice were identified through genotyping using Southern blotting analysis. Both heterozygous and homozygous UPII-SV40T mice were fed orally with vehicle control or 6 g flavokawain A /kg food for 9 months starting at 6 weeks of age. More than 54% control food- fed male mice died of ureter obstruction and nephrohydrosis within 10.5 months of age, while more than 67% of mice fed with 6g flavokawain A /kg food survived beyond 10.5 months of age. The mean bladder weight of male transgenic mice that were fed with 6g flavokawain /kg food for 9 months decreased by about 46% compared to those fed with vehicle control diet (mean bladder weights for vehicle control versus flavokawain A treatments are 0.2031±0.016 versus 0.1108±0.02565, P=0.0037). The mean bladder weights of female transgenic mice that were fed with 6g flavokawain /kg food for 9 months decreased by 31% compared to those fed with vehicle control diet (mean bladder weights for vehicle control versus flavokawain A treatments are 0.0466±0.0052 versus 0.03225±0.004689, p=0.0556). Mouse pathology analysis revealed that vehicle control treated mice exhibited poorly differentiated transitional cell carcinoma in the bladder, while that flavokawain A treated mice presented well differentiated tumors in the bladder. The bladder tissues from flavokawain A treated mice showed a decreased number of Ki67 positive cells compared to those in vehicle control treated mice, which suggested that flavokawain A inhibited cell proliferation in vivo in bladder tumors. These results suggested a potential of flavokawain A in preventing the recurrence of non-muscle invasive urinary bladder cancer by treating CIS.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 619. doi:1538-7445.AM2012-619
Abstract 623: Prostate cancer and B-vitamins
Authors: Vidal Adriana , Grant Delores , Williams Christina , Allott Emma , Masko Elizabeth , Freedland Stephen J. , Hoyo Cathrine
Published: August 2012

BACKGROUND. Recent reports suggest that excess of nutrients involved in the one-carbon metabolism pathway increases PC risk, however empirical data are lacking. Our hypothesis was to examine whether excessive intake of one carbon nutrients was associated with PC risk and whether this association was similar in LG vs. HG prostate cancer. METHODS. Veteran American men (272 controls and 144 PC cases) who attended the Durham Veteran American Medical Center, between 2004-2011, were enrolled into a case-control study. Intake of folate, vitamin B12, B6 and methionine were measured using a food frequency questionnaire. Regression models were used to evaluate the association among one-carbon cycle nutrients, MTHFR genetic variants and prostate cancer. RESULTS. Higher methionine intake was positively associated with PC risk, OR=2.1 95%CI 1.1-3.9, p=0.02; a risk which increased for low grade PC or Gleason <7, OR=2.75 (95%CI 1.32-5.73), p=0.007. These associations did not vary by MTHFR C677T genotype, however the cross-product term for the association of MTHFR A1298C variant and methionine was statistically significant, compared to non-carriers (p=0.045). Evidence for associations between B vitamins (folate, B12 and B6) and PC risk were less clear. CONCLUSIONS. Our results suggest that high methionine intake increases PC risk. Higher intakes of folate and vitamin B6 may be protective against advanced prostate cancer- however the associations were weak, while vitamin B12 higher intakes showed a trend for significance for higher PC risk. Individuals carrying MTHFR A1298C variants, and consuming more than the recommended levels of methionine, may be at higher risk for PC. Key words: Prostate cancer (PC), folate, vitamins B6, B12, methionine, methylenetetrahydrofolate reductase (MTHFR).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 623. doi:1538-7445.AM2012-623