bluuebunny/crossref_metadata_2025 huggingface dataset:
Relation of Epicardial Fat Thickness with Carotid Intima-Media Thickness in Patients with Type 2 Diabetes Mellitus
Authors: Cetin Mustafa , Cakici Musa , Polat Mustafa , Suner Arif , Zencir Cemil , Ardic Idris
Published: May 2013

Aims.The aim of this study was to investigate the relationship of echocardiographic epicardial fat thickness (EFT) with carotid intima-media thickness (CIMT), in patients with type 2 diabetes mellitus (T2DM).Methods and Results.A total of 139 patients with T2DM (mean age 54.3 ± 9.2 and 49.6% male) and 40 age and sex-matched control subjects were evaluated. Echocardiographic EFT and ultrasonographic CIMT were measured in all subjects. Patients with T2DM had significantly increased EFT and CIMT than those of the controls (6.0 ± 1.5 mm versus 4.42 ± 1.0 mm,P<0.001and 0.76 ± 0.17 mm versus 0.57 ± 0.14 mm,P<0.001, resp.). EFT was correlated with CIMT, waist circumference, BMI, age, duration of T2DM, HbA1c in the type 2 diabetic patients. Linear regression analysis showed that CIMT (β=3.52,t=3.72,P<0.001) and waist circumference (β=0.36,t=2.26,P=0.03) were found to be independent predictors of EFT. A cutoff high risk EFT value of 6.3 mm showed a sensitivity and specificity of 72.5% and 71.7%, respectively, for the prediction of subclinical atherosclerosis.Conclusion.We found that echocardiographic EFT was significantly higher in patients with T2DM. Our study also showed that EFT was strongly correlated with waist circumference and CIMT as being independent of sex.
Inclusion Complex of Zerumbone with Hydroxypropyl-β-Cyclodextrin Induces Apoptosis in Liver Hepatocellular HepG2 Cells via Caspase 8/BID Cleavage Switch and Modulating Bcl2/Bax Ratio
Authors: Muhammad Nadzri Nabilah , Abdul Ahmad Bustamam , Sukari Mohd Aspollah , Abdelwahab Siddig Ibrahim , Eid Eltayeb E. M. , Mohan Syam , Kamalidehghan Behnam , Anasamy Theebaa , Ng Kuan Beng , Syam Suvitha , Arbab Ismail Adam , Rahman Heshu Sulaiman , Ali Hapipah Mohd
Published: May 2013

Zerumbone (ZER) isolated fromZingiber zerumbetwas previously encapsulated with hydroxypropyl-β-cyclodextrin (HPβCD) to enhance ZER’s solubility in water, thus making it highly tolerable in the human body. The anticancer effects of this new ZER-HPβCD inclusion complex via apoptosis cell death were assessed in this study for the first time in liver hepatocellular cells, HepG2. Apoptosis was ascertained by morphological study, nuclear stain, and sub-G1 cell population accumulation with G2/M arrest. Further investigations showed the release of cytochrome c and loss of mitochondrial membrane potential, proving mitochondrial dysfunction upon the ZER-HPβCD treatment as well as modulating proapoptotic and anti-apototic Bcl-2 family members. A significant increase in caspase 3/7, caspase 9, and caspase 8 was detected with the depletion of BID cleaved by caspase 8. Collectively, these results prove that a highly soluble inclusion complex of ZER-HPβCD could be a promising anticancer agent for the treatment of hepatocellular carcinoma in humans.
Performance Evaluation of IEEE 1609 WAVE for Vehicular Communications
Authors: Wen Jyh-Horng , Weng Chien-Erh
Published: May 2013

In IEEE 1609, it uses IEEE 802.11 Enhanced Distributed Channel Access (EDCA) mechanism to access the channel. IEEE 802.11 EDCA is a new wireless technology for wireless access in the vehicular environment (WAVE). It defines a new supplement to the existing IEEE 802.11 MAC protocol. In IEEE 802.11 EDCA, the aim is providing a QoS support. While the system serves different access categories (ACs), EDCA does not perform well under high load conditions. In order to improve the efficiency, we pay attention to the EDCA with transmit opportunity (TXOP) mechanism. We first proposed a Markov chain model and studied the behavior. We extend the model to support IEEE 802.11 EDCA and presented a more accurate analysis under nonideal channel environment. We also compared it with that without TXOP mechanism under channel error environment.
Molecular Profiling Predicts the Existence of Two Functionally Distinct Classes of Ovarian Cancer Stroma
Authors: Lili Loukia N. , Matyunina Lilya V. , Walker L. DeEtte , Benigno Benedict B. , McDonald John F.
Published: May 2013

Although stromal cell signaling has been shown to play a significant role in the progression of many cancers, relatively little is known about its importance in modulating ovarian cancer development. The purpose of this study was to investigate the process of stroma activation in human ovarian cancer by molecular analysis of matched sets of cancer and surrounding stroma tissues. RNA microarray profiling of 45 tissue samples was carried out using the Affymetrix (U133 Plus 2.0) gene expression platform. Laser capture microdissection (LCM) was employed to isolate cancer cells from the tumors of ovarian cancer patients (Cepi) and matched sets of surrounding cancer stroma (CS). For controls, ovarian surface epithelial cells (OSE) were isolated from the normal (noncancerous) ovaries and normal stroma (NS). Hierarchical clustering of the microarray data resulted in clear separations between the OSE, Cepi, NS, and CS samples. Expression patterns of genes encoding signaling molecules and compatible receptors in the CS and Cepi samples indicate the existence of two subgroups of cancer stroma (CS) with different propensities to support tumor growth. Our results indicate that functionally significant variability exists among ovarian cancer patients in the ability of the microenvironment to modulate cancer development.
Lazer-Leach Type Conditions on Periodic Solutions of Liénard Equation with a Deviating Argument at Resonance
Authors: Wang Zaihong
Published: May 2013

We study the existence of periodic solutions of Liénard equation with a deviating argumentx′′+f(x)x'+n2x+g(x(t-τ))=p(t),wheref,g,p:R→Rare continuous andpis2π-periodic,0≤τ<2πis a constant, andnis a positive integer. Assume that the limitslimx→±∞g(x)=g(±∞)andlimx→±∞F(x)=F(±∞)exist and are finite, whereF(x)=∫0x‍f(u)du. We prove that the given equation has at least one2π-periodic solution provided that one of the following conditions holds:2cos(nτ)[g(+∞)-g(-∞)]≠∫02π‍p(t)sin(θ+nt)dt, for allθ∈[0,2π],2ncos(nτ)[F(+∞)-F(-∞)]≠∫02π‍p(t)sin(θ+nt)dt, for allθ∈[0,2π],2[g(+∞)-g(-∞)]-2nsin(nτ)[F(+∞)-F(-∞)]≠∫02π‍p(t)sin(θ+nt)dt, for allθ∈[0,2π],2n[F(+∞)-F(-∞)]-2sin(nτ)[g(+∞)-g(-∞)]≠∫02π‍p(t)sin(θ+nt)dt, for allθ∈[0,2π].
New OTRA-Based Generalized Impedance Simulator
Authors: Gupta Ashish , Senani Raj , Bhaskar D. R. , Singh A. K.
Published: May 2013

Operational transresistance amplifier (OTRA) has attracted considerable attention in the recent literature in several applications such as impedance simulation, universal biquad filter realization, realization of sinusoidal oscillators and multivibrators. However, to the best knowledge of the authors, any OTRA-based generalized impedance simulator circuits have not been reported so far. The purpose of this paper is to present such a circuit.
Fibroblast Growth Factor Receptor 3 Is a Rational Therapeutic Target in Bladder Cancer
Authors: Gust Kilian M. , McConkey David J. , Awrey Shannon , Hegarty Paul K. , Qing Jing , Bondaruk Jolanta , Ashkenazi Avi , Czerniak Bogdan , Dinney Colin P. , Black Peter C.
Published: May 2013

Activating mutations of fibroblast growth factor receptor-3 (FGFR3) have been described in approximately 75% of low-grade papillary bladder tumors. In muscle-invasive disease, FGFR3 mutations are found in 20% of tumors, but overexpression of FGFR3 is observed in about half of cases. Therefore, FGFR3 is a particularly promising target for therapy in bladder cancer. Up to now, most drugs tested for inhibition of FGFR3 have been small molecule, multityrosine kinase inhibitors. More recently, a specific inhibitory monoclonal antibody targeting FGFR3 (R3Mab) has been described and tested preclinically. In this study, we have evaluated mutation and expression status of FGFR3 in 19 urothelial cancer cell lines and a cohort of 170 American patients with bladder cancer. We have shown inhibitory activity of R3Mab on tumor growth and corresponding cell signaling in three different orthotopic xenografts of bladder cancer. Our results provide the preclinical proof of principle necessary to translate FGFR3 inhibition with R3Mab into clinical trials in patients with bladder cancer. Mol Cancer Ther; 12(7); 1245–54. ©2013 AACR.
Abstract A40: Association between smoking status and free, total, and percent free prostate-specific antigen
Authors: Li Jun , Thompson Trevor , Joseph Djenaba , Master Viraj
Published: May 2013

Background: There are scant data available on the relationship between smoking and total prostate-specific antigen (tPSA), free PSA (fPSA) and percent-free PSA (%fPSA). Given the high prevalence of smoking and frequency of PSA screening, it is important to determine any association between smoking and PSA values using nationally representative data.

Materials and Methods: Included in the final study population were 3,820 men aged ≥40 years who participated in the 2001-2006 National Health and Nutrition Examination Survey and met eligibility criteria for PSA testing. The distributions of tPSA, fPSA, and %fPSA were estimated by sociodemographic and clinical characteristics. Multivariate linear regression models were fit to determine the adjusted relationship between smoking and tPSA and %fPSA while controlling for these characteristics simultaneously.

Results: For all ages combined, the median tPSA and fPSA were 0.90 (0.81–0.90) ng/ml and 0.26 (0.25–0.28) ng/ml, respectively. The multivariate linear regression analysis showed that tPSA was 7.9% and 12.2% lower among current and former smokers, respectively, than that among never smokers. High body mass index and diabetes were also statistically significantly associated with lower tPSA level. About one third of the men had a %fPSA<25. Current smokers had significantly lower %fPSA compared with former smokers.

Conclusions: Our findings that smoking is inversely associated with tPSA may have potential implications for the interpretation of PSA levels in men who are current or former smokers. Given the high prevalence of smoking, obesity, and diabetes, additional research on the combined effect of these health risk factors is warranted.

Citation Format: Jun Li, Trevor Thompson, Djenaba Joseph, Viraj Master. Association between smoking status and free, total, and percent free prostate-specific antigen [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A40.
Abstract A47: Akt mediates prostate cancer cell migration and invasion via affinity modulation of integrin αvβ3
Authors: Goc Anna , Liu Junxiu , Byzova Tatiana , Shenoy Somanath
Published: May 2013

Purpose: Prostate cancer cells often metastasize to the bones. The ability of prostate cancer cells to attach to several matrix proteins, migrate with a sense of direction in response to stimuli, invade to the adjacent tissue and enter the micro-circulation etc. are pre-requisites for them to metastasize to distant tissues. During this sojourn, prostate cancer cells keep transforming itself from one type to another by constantly changing the expression profile of its surface receptors and secreted factors, which in turn, helps them to respond to the cues from the micro-environment and mediate tumor progression. The current study is focused on the role of Akt kinase in the affinity modulation of integrin αvβ3 leading to prostate tumor invasion and metastasis.

Methods: We used a number of tumor cell functional assay methods, molecular biology and biochemistry tools, immunocytochemistry, electric cell impedance sensing technology etc. to establish a correlation between Akt activity and inside-out activation (affinity modulation) of integrin αvβ3 leading to prostate cancer cell invasion and metastasis.

Results: Our previous studies have shown that Akt is necessary for the affinity modulation of integrins in many normal cell types. Our results also implicated that Akt is necessary for prostate cancer cell functions such as proliferation, survival, invasion and migration. Current results revealed the involvement of Akt-mediated activation of integrin αvβ3 in human prostate cancer cells in vitro. We showed that Akt and integrin αvβ3 cooperation is required not only for tumor growth but also for invasion and further micro-metastasis to the blood vessels and eventually metastasis to the bone.

Conclusion: Our study indicated a causal relationship between Akt activity, expression of integrin αvβ3 (that are not at all expressed by normal prostatic epithelial cells) and its inside-out activation (affinity modulation). We also conclude that Akt-mediated activation of integrin αvβ3 is necessary to initiate the switch in prostate cancer cells to invade and metastasis to distant tissues such as bone. Our study projects Akt and integrin αvβ3 as potential therapeutic targets for prostate cancer therapy.

Citation Format: Anna Goc, Junxiu Liu, Tatiana Byzova, Somanath Shenoy. Akt mediates prostate cancer cell migration and invasion via affinity modulation of integrin αvβ3 [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A47.
Abstract A49: Characterization of pre-B-cell leukemia transcription factor 1 in prostate cancer
Authors: Ramberg Hakon , Alshbib Ayham , Berge Viktor , Wanzhong Wang , Svindland Aud , Taskén Kristin Austlid
Published: May 2013

The main purpose of our project is to determine the role of Pre-B Cell Leukemia Transcription Factor 1 (PBX1) in prostate cancer development and progression.

In this study of PBX1, we used immunohistochemical staining of benign and malignant prostate tissue as well as immunoblotting and Real-Time RT-PCR of different prostate cancer cell lines to characterize the expression pattern and regulation of PBX1. Furthermore, we transfected LNCaP with short interference RNA (siRNA) against PBX1 to determine its role in proliferation and differentiation of prostate cancer cells.

Our analyses of PBX1 expression in prostate cancer cell lines indicate that the level of PBX1 is higher in LNCaP and VCaP compared to LNCaP C4-2B and DU145, at both the mRNA and protein levels. Immunohistochemical analysis of PBX1 showed that the protein is mainly expressed in basal cells of benign tissue and in a minor sub-fraction of cells located in the luminal compartment. Tumor tissue lacks basal cells and the expression level of PBX1 was therefore down-regulated in malignant compared to benign tissue. In less differentiated tumor areas, the frequency of PBX1 positive cells was lower than in highly differentiated tumors. Thus, an inverse correlation between PBX1 staining and Gleason grade was observed. The TMAstudy included 42 patients radically operated for prostate cancer in 2004 (108 spots) and no significant correlations between PBX1 expression and clinical parameters were observed.

Cyclic AMP has previously been shown to induce growth arrest and neurite outgrowth of prostate cancer cells. We have shown that the level of PBX1 mRNA is down-regulated by cAMP. Transient transfection of PBX1 siRNA in LNCaP cells mimicked the morphological changes observed when LNCaP cells are stimulated with cAMP (forskolin) and both treatments reduced the growth rate of LNCaP cells. Several cell cycle regulated genes have been identified as cAMP target genes in LNCaP cells and we are currently investigating whether these are PBX target genes. Furthermore, preliminary studies indicate decreased expression of luminal differentiation markers in LNCaP cells transfected with PBX1 siRNA.

Based on these preliminary studies, we suggest that PBX1 might play a role in regulation of development and/or progression of prostate cancer, as suggested for other types of cancers, like ovarian and renal cancers.

Citation Format: Hakon Ramberg, Ayham Alshbib, Viktor Berge, Wang Wanzhong, Aud Svindland, Kristin Austlid Taskén. Characterization of pre-B-cell leukemia transcription factor 1 in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A49.
Abstract A58: Galbanic acid isolated from Ferula assafoetida exerts in vivo antitumor activity in association with antiangiogenesis and antiproliferation
Authors: Kim Sunghoon , Kim Kwanhyun , Jeong Soojin , Lee Hyojeong , Lu Junxuan
Published: May 2013

To investigate whether galbanic acid (GBA) exerts antiangiogenic and anticancer activities. Using human umbilical vein endothelial cell (HUVEC) model, we analyzed effects of GBA on cellular and molecular events related to angiogenesis. We tested its direct antiproliferative action on mouse Lewis lung cancer (LLC) cells and established its in vivo antiangiogenic and antitumor efficacy using LLC model.

GBA significantly decreased vascular endothelial growth factor (VEGF)-induced proliferation and inhibited VEGF-induced migration and tube formation of HUVECs. These effects were accompanied by decreased phosphorylation of p38-mitogen-activated protein kinase (MAPK), c-jun N-terminal kinase (JNK), and AKT, and decreased expression of VEGFR targets endothelial nitric oxide synthase (eNOS) and cyclin D1 in VEGF-treated HUVECs. GBA also decreased LLC proliferation with an apparent G2/M arrest, but did not induce apoptosis. In vivo, inclusion of GBA in Matrigel plugs reduced VEGF-induced angiogenesis in mice. Galbanic acid given by daily i.p. injection (1 mg/kg) inhibited LLC-induced angiogenesis in an intradermal inoculation model and inhibited the growth of s.c. inoculated LLC allograft in syngenic mice. Immunohistochemistry revealed decreased CD34 microvessel density index and Ki-67 proliferative index in GBA-treated tumors.

Overall, our findings suggest that GBA exerts anticancer activity in association with antiangiogenic and antiproliferative actions.

Citation Format: Sunghoon Kim, Kwanhyun Kim, Soojin Jeong, Hyojeong Lee, Junxuan Lu. Galbanic acid isolated from Ferula assafoetida exerts in vivo antitumor activity in association with antiangiogenesis and antiproliferation [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A58.
Abstract A61: Hexim-1 protein expression levels modulate TGFβ activity in prostate cancer
Authors: Dhar-Mascareno Manya , Mascareno Eduardo , Liu Fang
Published: May 2013

Hexim-1 is a highly conserved protein from human to mouse, and a well-established inhibitor of RNA polymerase II-dependent transcription elongation and cell proliferation. Others and we have proposed and provided evidence that changes in the expression and activity in Hexim-1 significantly influence the progression of cancer. Here we show that changes in expression levels of Hexim-1 modulate TGFβ1 signaling by inducing posttranslational modifications in the SMADs proteins (small mother against decapentaplegic) at the polylinker region of the Smad1 and Smad3 proteins. Cdk9 serine activity not only targets specific serine residues in the carboxyl terminus of RNA polymerase II during transcriptional elongation but also serine residues in the polylinker region of Smad3. Hexim-1 directly inhibits Cdk9 activity, and therefore, transcription elongation and TGFβ1/Smad3 activity. Our results indicate that prostate cancer accompanies significant changes in Hexim-1 expression as well as sub cellular redistribution. Similar observations were obtained in the TRAMP mouse model of prostate cancer. In order to understand the role of Hexim-1 during prostate cancer progression, we generated a TRAMP mouse model heterozygous for Hexim-1. Our studies in the TRAMP mouse model revealed that lowered levels in Hexim-1 protein expression triggered a more aggressive form of prostate cancer. TGFβ1/Smad3 plays a significant role during proliferation, cell migration, as well as in epithelial mesenchymal transition, which is critical during metastasis. To test the role of Hexim-1 as a modulator of TGF/Smad3 in vitro, we use the epithelial mouse prostate cancer cell line TRAMP-C2 and derived the TRAMP-C2 heterozygous for Hexim-1 (HT). We observed increased serine 208 phosphorylation of Smad3 as well as enhanced transcriptional activity of TGF β/Smad3 target genes involved in EMT such as SM22alpha and alpha smooth muscle actin in the HT-TRAMP-C2 cell line. These findings suggest that Hexim-1 is a critical regulator of prostate cancer progression.

Citation Format: Manya Dhar-Mascareno, Eduardo Mascareno, Fang Liu. Hexim-1 protein expression levels modulate TGFβ activity in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr A61.
Abstract B10: Inactivation of androgen-induced coactivator ARD1 inhibits androgen receptor acetylation and prostate tumorigenesis
Authors: Wang Zehua , Wang Zemin , Guo Jianhui , Liu Wanguo
Published: May 2013

Androgen signaling through androgen receptor (AR) is critical for prostate tumorigenesis. Given that AR-mediated gene regulation is enhanced by AR coactivators, inactivation of those coactivators is emerging as a promising therapy for prostate cancer (PCa). Here, we show that the N-acetyltransferase Arrest-defect-1 protein (ARD1) functions as a novel AR coactivator in PCa cells. ARD1 is upregulated in human PCa cell lines, tumor biopsies, and in prostate tumor tissues of Pten null mouse. The expression of ARD1 was augmented by treatment with synthetic androgen (R1881) unless AR is deficient or is inhibited by AR-specific siRNA or androgen inhibitor bicalutamide (Casodex [CDX]). Depletion of ARD1 by shRNA suppressed PCa cell proliferation, anchorage-independent growth, and xenograft tumor formation in SCID mice, suggesting that AR-dependent ARD1 expression is biologically germane. Notably, ARD1 was necessary and sufficient to transcriptionally coactivate a number of AR-target genes that are involved in prostate tumorigenesis shown by qRT-PCR, reporter assay, and chromatin immunoprecipitation (ChIP) analysis. Furthermore, ARD1 but not ARD1-dead (acetylase activity null mutant) interacted physically with and acetylated the AR protein in vivo and in vitro. Moreover, the ARD1-mediated AR acetylating is p300 independent. Since ARD1-specific AR acetylation facilitated the AR binding to its targeted promoters for gene transcription, we propose that ARD1 functions as a unique AR coactivator and forms a positive feedback loop for AR-dependent prostate tumorigenesis. Disruption of AR-ARD1 interactions may be a potent intervention for androgen-dependent PCa therapy.

Citation Format: Zehua Wang, Zemin Wang, Jianhui Guo, Wanguo Liu. Inactivation of androgen-induced coactivator ARD1 inhibits androgen receptor acetylation and prostate tumorigenesis [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B10.
Abstract B43: An accurate seven-gene signature plus Gleason score for prostate cancer prognosis
Authors: Chen Xin , Jia Zhenyu , Dan Mercola
Published: May 2013

One of the major problems in the development of prostate cancer prognostic biomarkers is the composition heterogeneity in tissue samples, which causes the inconsistency among different studies. To deal with this problem, we developed two-step strategy. In the first step, the proposed two-step cluster analysis model accounting for the cell-type composition percentage was used to identify 324 genes strongly associated with biochemical recurrence status and in the second step the 324 identified genes was further investigated by SAM (significance analysis of microarray) leading to a seven-gene classifier. The classifier was then validated on two independent data sets with overall accuracy and sensitivity of 71% and 76%, respectively. The seven-gene classifier stratified the test samples into two risk groups that are highly associated with biochemical relapse status (hazard ratio = 2.6 with p value = 0.03). The study also showed that the inclusion of the Gleason sum to the seven-gene classifier raised the sensitivity and accuracy to 76% and 83% respectively when tested on the independent data sets. These results indicated that our seven-gene classifier with the Gleason score can be used for predicting outcomes for prostate cancer patients.

Citation Format: Xin Chen, Zhenyu Jia, Mercola Dan. An accurate seven-gene signature plus Gleason score for prostate cancer prognosis [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B43.
Abstract B5: Epigenetic modification of AR in PC3 cells negatively regulates mTORC1/2 activity
Authors: Ellis Leigh , Adelaiye Remi , Ku Shengyu , Godoy Alejandro , Pili Roberto
Published: May 2013

Wild type androgen receptor (AR) acts as a tumor suppressor and mediates the differentiation of non-malignant prostate epithelial cells. However, in the development of prostate cancer AR activity becomes critical for the development and progression of disease. Previous studies demonstrate that re-expressing wild type AR within an AR negative cell line (PC3) possesses tumor suppressor quality by reducing cell proliferation and tumorgenicity in mice. Further, it is documented that acetylation of AR in non-malignant cells is a post-translational modification associated with increased transcriptional activity of AR, whereas acetylation of AR in tumor cells results in negative regulation of AR transcriptional activity. We therefor are seeking to investigate the regulation of re-expressed AR within PC3 cells by induced acetylation post HDAC inhibition. Our preliminary data reveals that sensitivity to HDAC inhibition is increased with the re-expression of AR in PC3 cells. Further, epigenetic modification of AR in this model negatively regulates mTORC1 and mTORC2 expression and activity, independent of AR transcriptional activity. Ongoing experiments are being conducted to investigate further the role of epigenetic post-translational modification of AR and its negative regulation of mTORC1/2 signaling.

Citation Format: Leigh Ellis, Remi Adelaiye, Shengyu Ku, Alejandro Godoy, Roberto Pili. Epigenetic modification of AR in PC3 cells negatively regulates mTORC1/2 activity [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B5.
Abstract B52: Loss of anterior gradient protein in high-grade prostate cancer is associated with loss of tumor cell adhesion and epithelial cell polarity
Authors: Chanda Diptiman , Sawant Anandi , Hensel Jonathan Adam , Isayeva Tatyana , Reilly Stephanie D. , Siegal Gene P. , Smith Claire , Ponnazhagan Selvarangan
Published: May 2013

Prostate cancer has the highest rate of incidence among all cancers in men in the United States and the second leading cause of death behind lung cancer. The human homologue of the frog (Xenopus) anterior gradient protein (AGR-2) has gained considerable attention in recent years for its putative role in neoplastic growth, metastasis and poor survival of patients with breast and prostate cancers. AGR-2 is reported to be androgen dependent and expressed only during the early stages of prostate cancer progression, whereas significant decline in AGR-2 levels in higher grades tumors coincided with metastatic disease. Human primary prostate cancer tissues and regional metastasis were immunostained for AGR-2 expression. Results indicated that AGR-2 was localized in the glandular epithelium of the prostate gland. No significant difference in AGR-2 staining was observed between normal, BPH and well-differentiated prostate cancers, except in normal prostate, where AGR-2 was mostly concentrated in the nuclei. Similar results were observed in a spontaneously developing transgenic adenocarcinoma of mouse prostate (TRAMP) model. AGR-2 expressions was reduced significantly in moderately and poorly-differentiated tumors in both human and TRAMP mice. Higher AGR-2 levels were found in the metastatic tissue albeit lower expressions in the highest grades prostate tumors within the same individual, suggesting further requirement of AGR-2 for growth at the metastatic sites. Targeted silencing of AGR-2 in a bone metastatic prostate cancer cell line, PC3, resulted in significantly reduced cellular adhesion to various extracellular matrix proteins and a failure to form acinar structures following 3-dimensional growth on matrigel. We also determined that AGR-2 silencing led to down-regulation of crucial integrins in PC3 cells, which possibly explains mechanism by which AGR-2 maintains polarized morphology of epithelial cells through promotion of cell adhesion. Based on our findings, we hypothesize that loss of AGR-2 in prostate cancer cells allows detachment from the basement membrane of the primary site and initiates metastasis. AGR-2 as well as integrin levels are restored during passage through circulation and at the metastatic site(s), helping them re-attach to the new microenvironment.

Citation Format: Diptiman Chanda, Anandi Sawant, Jonathan Adam Hensel, Tatyana Isayeva, Stephanie D. Reilly, Gene P. Siegal, Claire Smith, Selvarangan Ponnazhagan. Loss of anterior gradient protein in high-grade prostate cancer is associated with loss of tumor cell adhesion and epithelial cell polarity [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B52.
Abstract B57: Oncostatin M promotes progression of Pten-deficient prostate cancer
Authors: Smith Daniel A. , Zong Yang , Kiba Atsushi , Witte Owen N.
Published: May 2013

Chronic inflammation has been proposed to play a crucial role in several cancers including gastric, pancreatic, colon and lung, with recent evidence suggesting involvement in prostate cancer. While both interleukin 6 (IL6) and the related cytokine oncostatin M (OSM) have been implicated in prostate cancer, there have been few studies using in vivo models. To address this issue, we employed a dissociated prostate tissue recombination system using epithelium derived from conditional Pten knockout mice. Loss of Pten is common in prostate cancer and generates an intermediate murine prostatic intraepithelial neoplasia (mPIN) lesion in our system, providing an ideal means to assess functional synergy. We hypothesize that increased expression of these cytokines in the prostate microenvironment will promote malignant progression of intermediate prostate cancer lesions.

Paracrine expression of IL6 did not transform naïve prostate epithelium nor did it exhibit functional synergy with Pten null epithelium in our system. Grafts expressing OSM alone did not transform naïve tissue, though in contrast, presented dramatic functional synergy with loss of Pten in prostate epithelium. Loss of Pten combined with OSM exhibited progression to poorly differentiated adenocarcinoma with features of epithelial-to-mesenchymal transition (EMT). Transformed tissues displayed loss of cytokeratin expression with down-regulation and mis-localization of e-cadherin. These tumors exhibit activation of several pathways downstream of OSM signaling including STAT3 and Erk1/2 as seen by immunohistochemical analysis. Further, we interrogated the role of OSM in promoting hormone refractory prostate cancer by castrating host animals following tumor establishment. Loss of Pten alone failed to progress beyond mPIN lesions and eventually regressed at 6 months post castration. In dramatic contrast, grafts expressing the OSM ligand exhibited initial regression with castration resistant growth following a latency period, recapitulating events commonly observed in human disease.

Our evidence suggests that in the context of Pten loss, IL6 does not appear to promote prostate malignancy in the murine system. Conversely, we have identified that OSM synergizes with loss of Pten to promote aggressive prostate malignancy. Paracrine expression of OSM results in activation of several pathways, providing a means to interrogate hierarchy within signaling modules in the context of prostate cancer progression. Our study further indicates that OSM could play a role in promoting castration resistance, allowing investigation of potential points for therapeutic intervention.

Citation Format: Daniel A. Smith, Yang Zong, Atsushi Kiba, Owen N. Witte. Oncostatin M promotes progression of Pten-deficient prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B57.
Abstract B67: The epigenetic regulator Hmga2 is sufficient to initiate prostate cancer via paracrine Wnt signaling
Authors: Zong Yang , Huang Jiaoti , Witte Owen N.
Published: May 2013

Carcinomas result from the stepwise acquisition of multiple genetic alterations within the epithelial compartment. The surrounding stroma, including extracellular matrix, blood vasculature, inflammatory cells and fibroblasts, also plays important roles in cancer initiation and progression. Aberrant microenvironment resulting from aging and chronic inflammation as well as tumor stress can provide “field effects” to facilitate the development of multifocal malignant lesions with independent clonal origin. Given the rare frequency of genetic changes identified in cancer associated stroma, it is likely that epigenetic alterations, such as histone modifications and chromatin remodeling, could contribute to the tumor-promoting phenotype of cancer associated stromal cells. Here we utilize high-mobility group AT-hook 2 (Hmga2), a nonenzymatic epigenetic regulator, to modify prostate stromal cells, and demonstrate that perturbation of the microenvironment by stromal-specific overexpression of this chromatin remodeling protein is sufficient to induce multiple high-grade prostatic intraepithelial neoplasia lesions from adjacent naïve epithelial cells. The growth-promoting effects of Hmga2-expressing stromal cells are predominantly mediated by increased Wnt/β-catenin signaling and preferentially act on prostate stem cells-enriched basal epithelial cells. Enhancement of Hmga2-induced paracrine signaling by overexpression of androgen receptor (AR) in the stroma drives full carcinoma in the adjacent epithelial tissues, indicating that combined targeted therapies against augmented Wnt and stromal AR signaling may represent as an effective treatment strategy for prostate cancer. Our findings provide compelling evidence for the critical contribution of epigenetic changes in stromal cells to multifocal tumorigenesis, and for the first time reveal that Hmga2, a cell intrinsic oncoprotein, could exert its function via paracrine Wnt signaling.

Citation Format: Yang Zong, Jiaoti Huang, Owen N. Witte. The epigenetic regulator Hmga2 is sufficient to initiate prostate cancer via paracrine Wnt signaling [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr B67.
Abstract C11: Using GCP to improve initial response to androgen deprivation in prostate cancer
Authors: Mooso Benjamin A. , Mudryj Maria , deVere White Ralph W. , Ghosh Paramita M. , Savoy Rosalinda M. , Cheung Jean P. , Vinall Ruth L. , Tepper Clifford G. , Wang Yu , Siddiqui Salma , Bedolla Roble G. , Mikhailova Margarita
Published: May 2013

Recurrent prostate cancer (PCa) is treated with androgen deprivation therapy (ADT, chemical castration), but patients on this treatment frequently relapse, indicative of the advent of castration resistant prostate cancer (CRPC). Due to the multifocal nature of the disease, any one treatment may not affect all pathways altered in CRPC; therefore, we set out to delay CRPC by increasing the apoptotic rate during ADT. We previously published that Filamin A (FlnA) is localized to the nucleus in androgen-dependent cells, whereas its expression is lost from the nucleus in CRPC. Restoration of FlnA nuclear expression induced apoptosis in the absence of androgens. We now show that genistein combined polysaccharide (GCP), a combination of isoflavones, can induce FlnA translocation to the nucleus. GCP and its main component genistein previously failed to prevent progression in pre-prostatectomy patients with low-grade PCa. However, our new data, both in vitro and in an animal model, show that GCP impedes relapse following castration, and induces apoptosis in PCa cells undergoing ADT, by promoting FlnA cleavage and nuclear translocation. Investigation of the mechanism of GCP's effects on FlnA showed that ADT phosphorylates FlnA and prevents its cleavage, whereas GCP inhibits FlnA phosphorylation, thereby promoting its cleavage and nuclear translocation. Additional data show that nuclear localization of FlnA induces apoptosis during ADT by preventing androgen receptor (AR) stabilization and by suppression of the ErbB3/PI3K/Akt axis. Taken together, our results identify GCP as a putative therapeutic module to prolong the effectiveness of ADT in patients with metastatic prostate cancer.

Citation Format: Benjamin A. Mooso, Maria Mudryj, Ralph W. deVere White, Paramita M. Ghosh, Rosalinda M. Savoy, Jean P. Cheung, Ruth L. Vinall, Clifford G. Tepper, Yu Wang, Salma Siddiqui, Roble G. Bedolla, Margarita Mikhailova. Using GCP to improve initial response to androgen deprivation in prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr C11.
Abstract C23: MicroRNA-106b targets caspase-7 and is associated with recurrence in human prostate cancer
Authors: Hudson Robert S. , Dorsey Tiffany H. , Yi Ming , Esposito Dominic , Stephens Robert M. , Croce Carlo M. , Ambs Stefan
Published: May 2013

Previously, we found several microRNA (miR) of the polycistronic miR-106b-25 cluster to be elevated in prostate cancer. This cluster is a homolog to the prototypical oncomiR-1 (miR-17-92) and is thought to act as a super-oncogene when co-amplified with its host gene minichromosome complex maintenance component 7 (MCM7). In this study we validated our previous findings in an independent prostate cancer dataset and further found that miR-106b is elevated in metastatic tissues and associated with disease recurrence. Genome-wide analysis of miR-106b-25 correlated transcripts revealed enrichment of focal adhesion and ECM-receptor interaction genes. Further, we found miR-106b target enrichment in transcripts that were protective against recurrence, suggesting that miR-106b targets collectively have a functionally protective role in the recurrence of prostate cancer. In cell based assays, overexpression of miR-106b increased proliferation, adhesion-free survival, soft agar survival, and adhesion to ECM components (laminin, and collagen type I and IV). In addition, microarray analysis showed that miR-106b down-regulated transcripts were enriched for miR-106b targets sites and cell death pathways genes. When we compared genes that were related to recurrence, altered by miR-106b, and predicted to have miR-106b 3′UTR target sites we identified CASP7, a central executioner of apoptosis. Corroborating these findings we found that the expression of CASP7 was consistently reduced in primary human prostate tumors and metastatic lesions across multiple prostate cancer datasets, and is by itself a candidate predictor of disease recurrence. Western blot analysis in RWPE-1, 22Rv1, and LNCaP prostate cells showed that miR-106b was capable of reducing CASP7 protein levels. Direct targeting of the CASP7 3′UTR by miR-106b was confirmed using wild-type and mutant 3′UTR luciferase reporter assays. This research reveals a novel oncogenic role for the miR-106b-25 cluster in prostate cancer and, for the first time, indentifies CASP7 as a direct target of miR-106b. Future work is aimed at understanding the role of miR-106b-dependent regulation of CASP7 and the possible utility of targeting miR-106b as novel therapy for prostate cancer.

Citation Format: Robert S. Hudson, Tiffany H. Dorsey, Ming Yi, Dominic Esposito, Robert M. Stephens, Carlo M. Croce, Stefan Ambs. MicroRNA-106b targets caspase-7 and is associated with recurrence in human prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr C23.
Abstract C61: The use of beta blockers improve cancer-specific survival in prostate cancer patients receiving androgen deprivation therapy
Authors: Grytli Helene Hartvedt , Håheim Lise Lund , Taskén Kristin Austlid
Published: May 2013

Objective: To examine the potential effects of beta blocker use on clinical prognostic factors present at the time of prostate cancer diagnosis and on prostate cancer specific survival.

Experimental procedures: The present study is a prospective study of men who participated both in the Oslo study of 1972/73 and in a follow-up health screening in 2000 (n=6515). Self-administrated questionnaires provided information about use of beta blockers during the past four weeks before the survey in 2000 (n=776; 11.9%). Data from the 2000 survey were coupled with clinical and prognostic information from the Norwegian Cancer Registry to identify those men diagnosed with prostate cancer. Mortality data was obtained from the Norwegian Causes of Death Registry. All men diagnosed with prostate cancer that did not report the use of a beta blocker were considered the control group.

Results: Out of the 6515 men participating in the Oslo II study, a total of 655 men (10.1%) were diagnosed with prostate cancer. There was no significant differences between beta blocker users (n=80) and non users (n=575) regarding PSA, clinical T-stage, Gleason score, metastasis, or age at diagnosis. There were also no effects on prostate cancer specific survival in the group as a whole, analyzed by competing risk regression modeling (adjusted subdistribution hazard ratio (SHR) 0.69, 95% confidence interval (CI) 0.36-1.30, p = 0.252). When stratifying patients according to androgen deprivation therapy (ADT) from time of diagnosis, there was an association between beta blocker use and improved prostate cancer specific survival for men in the ADT- treated group (n=263), with an adjusted SHR of 0.18 (95% CI 0.03–0.96, p = 0.044). For patients not treated with ADT from the time of diagnosis (n=227), no difference on prostate cancer specific survival was seen (adjusted SHR 1.19, 95% CI 0.20-6.99, p = 0.844). Adjustment was made for metastasis, age at diagnosis and level of education as recorded in the 2000 screening.

Conclusion: The use of beta blockers was associated with an increased prostate cancer specific survival in men treated with ADT.

Citation Format: Helene Hartvedt Grytli, Lise Lund Håheim, Kristin Austlid Taskén. The use of beta blockers improve cancer-specific survival in prostate cancer patients receiving androgen deprivation therapy [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr C61.
Abstract C64: Adult murine prostate basal and luminal cells are self-sustained lineages that can both serve as targets for prostate cancer initiation
Authors: Choi Nahyun , Zhang Boyu , Zhang Li , Ittmann Michael , Xin Li
Published: May 2013

The prostate epithelial lineage hierarchy and the cellular origin for prostate cancer remain inadequately defined. We sought to utilize a lineage tracing approach to define the epithelial lineage hierarchy in the prostate. A previously reported K14-CreER transgenic mouse line was bred with a fluorescent reporter line to specifically label the prostate basal cell lineage. In parallel, the prostate luminal cell lineage was labeled with fluorescence using a novel K8-CreERT2 transgenic mouse line generated in our own lab. Using this lineage tracing approach, we show that adult rodent prostate basal and luminal cells are independently self-sustained in vivo.

We then sought to characterize the cells of origin for prostate cancer in the context of loss of function of Pten, which occurs frequently in primary and advanced human prostate cancer. We bred the K14-CreER and K8-CreERT2 mouse lines with Ptenfl/fl mice separately so that Pten expression can be disrupted specifically in prostate basal and luminal cells, respectively. Disrupting Pten in either lineage led to prostate cancer initiation. However, the cellular composition and onset dynamics of the resulting tumors are distinctive. Prostate luminal cells are more responsive to Pten null-induced mitogenic signaling. In contrast, basal cells are resistant to direct transformation. Instead, loss of Pten activity induces the capability of basal cells to differentiate into transformation-competent luminal cells.

These studies suggest that deregulation of the normal prostate epithelial differentiation program is a critical step for initiation of human prostate cancer with a basal cell origin. Furthermore, suppressing signaling pathways that induce basal-luminal differentiation may provide an efficient approach to prevent prostate cancer initiation.

Citation Format: Nahyun Choi, Boyu Zhang, Li Zhang, Michael Ittmann, Li Xin. Adult murine prostate basal and luminal cells are self-sustained lineages that can both serve as targets for prostate cancer initiation [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr C64.
Abstract IA1: COP1: Tumor suppressor role revealed
Authors: Dixit Vishva M.
Published: May 2013

Proto-oncogenes ETV1, ETV4, and ETV5 belong to the PEA3 family of ETS transcription factors and have been identified recently in prostate cancer chromosomal rearrangements that result in their overexpression. Little is known, however, about their post-translational regulation. Here we show that ETV1, ETV4, and ETV5 interact with the ubiquitin ligase COP1. COP1 together with its binding partner DET1 ubiquitinated ETV1, resulting in ETV1 proteasomal degradation. Importantly, we show that truncation of ETV1 by the TMPRSS2:ETV1 translocation found in prostate cancer removes its COP1 binding motif and thereby enhances its stability. Consistent with a critical role for COP1 in regulating PEA3 transcription factors, COP1 deficiency in primary mouse prostate cells produced prostate intraepithelial neoplasia in vivo. The relationship between COP1 and ETV1 was also evident in human prostate cancer samples where loss of COP1 expression correlated with elevated ETV1 protein.

Citation Format: Vishva M. Dixit. COP1: Tumor suppressor role revealed [abstract]. In: Proceedings of the AACR Special Conference on Advances in Prostate Cancer Research; 2012 Feb 6-9; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(4 Suppl):Abstract nr IA1.