bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 455: Phospho-ERK as a predictor for different ways of metastasis and / or survival in patients with metastasized colon cancer
Authors: Strohäker Jens , Falch Claudius , Weyhern Claus Hann von , Adam Patrick , Königsrainer Alfred , Fend Falko , Brücher Björn LDM
Published: August 2012

Background: Not much is known about different signalling pathways and its influence on metastasis in colon cancer. Phospho-ERK (pERK) is a downstream-target of K-Ras within the EGFR-Pathway. The goal of our investigations was the evaluation of the MAP-Kinase-cascade in regard to different ways of metastasis. Methods: 98 patients with adenocarcinom of the colon and a postoperative pT2/3-status, who underwent surgical resection between 2006 and 2008 at the University of Tübingen had been included within the study collective. 53 patients had no metastasis (control group), while 45 patients revealed primary metastasis: lymphogenetic metastasis (n=20), hepatic spread (n=20) and multiple metastasize (n=5). On a tissue-micro-Array pERK was stained by immunohistochemistry, the analysis was done semiquantitatively. Results: The graduation in immunohistochemistry was done as followed: negative staining (n=31), weak positive staining (n=36), moderate staining (n=20) and strong staining (n=11). 19 out of 31 negative patients (61%) revealed metastasis, while this had been 17 out of 36 weak positive (47%), 6 out of 20 (30%) moderate and 4 out of 11 (36%) strongly positive stained patients (p=0.229). No significant correlation was observed for lypmhogenetic (p=0.683) or multiple metastatic localization (p=0.666) and the pERK-expression. A significant correlation between the pERK-expression and the number of hepatic metastasis was observed (p=0.049). Additionally pERK-negative patients (31/98 = 32%) revealed a significant worse survival compared to those with positive pERK-staining (p=0.003). Conclusion: Our first results show, that a weak pERK-expression is correlated with a higher incidence in patients with liver metastasis. Patients without a pERK-expression revealed a worse survival compared to patients with pERK-expression.

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 455. doi:1538-7445.AM2012-455
Abstract 4553: Multiplexed, digital gene expression and fusion transcript analysis to screen for EML4-ALK positive lung cancer
Authors: Lira Maruja E. , Kim Tae-Min , Huang Donghui , Deng Shibing , Koh Youngil , Jang Bogun , Lee Se-Hoon , Chung Doo Hyun , Kim Woo Ho , Kim Dong-Wan , Mao Mao
Published: August 2012

Purpose: EML4-ALK fusions occur in approximately 5% of non-small cell lung carcinoma and define a subpopulation of lung cancer patients highly responsive to ALK kinase inhibitors. Current methodologies for detecting presence of ALK rearrangements are labor-intensive, costly, and not ideally suited for screening large number of patient samples. To allow for a sensitive, facile, and inexpensive methodology to detect EML4-ALK fusions, we developed a direct transcript profiling to detect common variants of EML4-ALK fusions. Experimental Design: Using a single multiplex assay, we simultaneously interrogated presence of EML4-ALK transcripts and ALK 3′ over-expression in 8 ALK-positive and 22 ALK-negative NSCLC samples previously tested by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). For controls, we also tested ALK-positive cancer cells lines NCI-H3122 and NCI-H2228, and an ALK-negative cell line A549. Purified RNA from formalin-fixed paraffin-embedded (FFPE) tissue sections was hybridized in duplicates to a biotinylated capture probe and a color-coded reporter probe designed to hybridize to sequences spanning EML4-ALK junction. In addition, multiple probes were created to hybridize within 5′ and 3′ regions of ALK transcript to detect discordant levels between the breakpoint, indicative of fusion event. Purified hybridization complexes are immobilized and aligned on a cartridge where a microscope CCD camera images and counts the molecular color-coded tags attached to reporter probes. The number of reporter counts corresponds to the number of transcripts being interrogated. Results: With this assay, we obtained 100% concordance in EML4-ALK fusion calls to results generated by FISH and IHC analyses. Similar findings were obtained for control cancer cell lines. The assay is highly reproducible and sensitive, detecting ALK-fusion transcripts even in samples with low tumor content. Using the combined strategy of fusion detection and ALK 5′ and 3′ transcript quantification, we were able to validate the status of one patient negative for ALK rearrangement by FISH but positive for ALK protein expression by IHC. Conclusions: We have developed a novel and sensitive methodology to screen for common EML4-ALK fusions in NSCLC. The assay is inexpensive, easy to perform, high-throughput and compatible with FFPE tissue samples. This is a promising technology highly suitable for screening large numbers of tumor samples without the need for cDNA synthesis and PCR amplification.

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 4553. doi:1538-7445.AM2012-4553
Abstract 4556: Epidermal growth factor signaling alterations in uterine papillary serous carcinoma (UPSC) associate with survival
Authors: Groeneweg Jolijn W. , Tambouret Rosemary H. , Borger Darrell R. , Foster Rosemary , Rueda Bo R. , Growdon Whitfield B.
Published: August 2012

Endometrial cancer is the most common gynecologic malignancy and the majority of patients are cured with surgery alone. A substantial subset of patients, however, present with high grade uterine papillary serous carcinoma (UPSC) and these patients account for a disproportionate degree of the recurrence, chemoresistance and death associated with endometrial cancer. Novel treatment strategies incorporating molecularly targeted therapies would be of great value in effective clinical management of these tumors. HER2 (ErbB2) hyperactivity has been associated with 10-20% of UPSC, though efforts to target this signature with trastuzumab have yielded no response in a recent prospective trial. In breast cancer, HER2/HER3 (ErbB2/ErbB3) dimerization has been associated with trastuzumab resistance. The expression profile of HER3 in UPSC is currently unknown. The objective of this investigation was to characterize HER2 and HER3 expression in a UPSC series and correlate these findings with clinical outcomes including survival. Under an IRB approved protocol, we identified 33 patients who underwent initial surgical staging at our institution between 2000 and 2009 with archival formalin fixed, paraffin embedded blocks and complete clinical follow up data. These tumor samples were subjected to immunohistochemistry (IHC) for HER2 and HER3 as well as fluorescent in situ hybridization (FISH) to assess the amplification status of the HER2 gene. Non-parametric tests were utilized to test correlation between detected levels of HER2 or HER3 protein expression and HER2 gene amplification and survival analysis was performed using the Kaplan-Meier method. HER2 gene amplification was observed in 34% of the cohort while HER2 protein over-expression, defined as 2+ or 3+ staining, was found in 59% of samples tested. HER2 gene amplification was associated with high HER2 protein expression (p < 0.05). Moreover, HER2 gene amplification was significantly associated with a decreased overall survival (p < 0.05). Finally, HER3 over-expression, defined as 2+ or 3+ staining, was observed in 52% of the entire cohort and HER3 protein levels exhibited a significant inverse relationship to HER2 protein expression and HER2 gene amplification (p < 0.05). These findings suggest that HER2 and HER3 signaling may function independently of one another in UPSC. The observed alterations in HER2 and HER3 expression levels constitute clinically relevant and prevalent signatures in UPSC that merit further investigation to determine the potential benefits of therapies targeting the epidermal growth receptor pathways.

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 4556. doi:1538-7445.AM2012-4556
Abstract 4558: FDG-PET uptake in breast cancer molecular subtypes
Authors: Elias Sjoerd G. , Wisner Dorota J. , Chen Yunn-Yi , Behr Spencer C. , Griffin Ann , Esserman Laura J. , van't Veer Laura J. , Hylton Nola M.
Published: August 2012

Background: Data on breast cancer FDG uptake in relation to clinically relevant molecular subtypes and their interaction is scarce. Some have shown higher standardized uptake values (SUV) in triple negative (TN) and HER2+ than hormone receptor (HR)+ disease. We hypothesize that FDG uptake in HER2+ disease depends on HR status. Methods: We identified 186 invasive breast cancer patients with a whole body FDG-PET/CT scan between 2005 and 2010 at UCSF (within 6 months prior to or after diagnosis but before therapy). Receptor expression was assessed on adjuvant therapy naïve specimens using IHC (HR [ER and PR], HER2) and FISH (HER2) according to clinical practice. We related routinely assessed (log-transformed) SUVmax to receptor expression with linear regression and analysis of covariance (ANCOVA), and adjusted for tumor grade. Results: SUVmax was reported for 160 primary breast cancers and 154 had complete data for HR and HER2. Median age at diagnosis was 52 years (range 21-87), and 34% had localized, 52% regional, and 14% distant disease. Average tumor size was 3.9 cm (SD 2.2), and 12% had good, 51% moderate and 37% poor grade. FDG uptake strongly depended on molecular subtype (Table). Uptake was lowest in HER2-HR+ and somewhat - but not significantly - higher in HER2+HR- disease. The highest SUVmax was found in HER2+HR+, followed by TN disease (both significantly higher than HER2-HR+). The interaction between HR and HER2 in relation to FDG uptake was significant (P=0.003). ANCOVA showed that the higher TN uptake could largely be explained by differences in grade, whereas SUVmax in HER2+HR+ disease remained significantly higher than in HER2-HR+ disease. Conclusion: This large series of advanced primary breast cancer showed FDG uptake to be comparably low in HER2-HR+ and HER2+HR- disease, but significantly higher in HER2+HR+ and TN disease. Tumor grade largely explained the higher FDG uptake in TN disease but not the results for HER2+HR+. FDG-PET may need evaluation at different settings taking into account uptake variation due to tumor biology.

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 4558. doi:1538-7445.AM2012-4558
Abstract 4559: A novel rotational Raman imaging device for early cancer detection
Authors: Garai Ellis , Zavaleta Cristina
Published: August 2012

A novel Raman imaging device that is capable of rotationally scanning hollow structures has been developed to be used in conjunction with tumor targeting Surface enhanced Raman scattering (SERS) nanoparticles. SERS nanoparticles can act as excellent contrast agents in that they offer ultra high sensitivity, and multiplexing capabilities due to the unique spectral signatures associated with them. One key advantage of multiplexing includes the ability to utilize several targeted SERS probes to target and localize a multitude of unique biomarkers associated with a specific disease, and effectively increasing cancer specificity. The device can potentially be used to distinguish a variety of tissues of interest in the colon such as: flat lesions, varying types of adenomas and carcinomas. There are also applications beyond the colon, such as the esophagus, stomach, oral cavity and bladder. In addition, multiplexing can allow for real-time functional imaging. A 45-degree scanning mirror, actuated with micromotor, is used to perform circumferential scans of the lumen of the colon at a rate of 1 revolution per second and a spatial resolution or approximately 1.5 mm. The device is able to be used with currently available endoscopes by inserting it into the working channel of an endoscope. The use of many (36) multi-mode fibers is used to increase the flexibility of the fiber bundle so that it can be inserted and used in a fully articulating endoscope while still being able to collect a sufficient amount scattered Raman light. When in the circumferential scanning mode, American National Standards Institute (ANSI) regulations will allow us to increase the laser illumination power to about 580 mW, resulting in a detection limit of approximately 1 pM of SERS nanoparticles. One of the key features of our Raman device is that it has been designed for efficient use over a wide range of working distances. This is necessary to accommodate for imperfect centering of the device within the colon, as well as variable working distances due to folds and bends within the colon. These techniques could allow endoscopists to distinguish between normal and cancerous tissues instantly, as well as to identify flat lesions that are easily missed during conventional screening endoscopy. After a suspicious region has been identified, the rotating mirror can be held at a certain angle, and used to help guide biopsies or resection of tissues in real-time.

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 4559. doi:1538-7445.AM2012-4559
Abstract 4562: Comprehensive analysis of the TMPRSS2-ERG translocation during prostate cancer progression
Authors: Ruiz Christian , Braun Martin , Scheble Veit J. , Zellweger Tobias , Rentsch Cyrill A. , Bachmann Alexander , Dodi Albert , Perner Sven , Sesterhenn Isabell A. , Srivastava Shiv , Bubendorf Lukas
Published: August 2012

Introduction. Approximately half of the diagnosed prostate carcinomas (PC) are characterized by a chromosomal rearrangement fusing the androgen regulated gene TMPRSS2 to the oncogenic ETS transcription factor ERG. Aim of this study was to comprehensively analyze the impact of this translocation on the expression of the ERG gene in hormone-naïve (untreated) and castration-resistant prostate cancers and to define the influence of AR protein expression and genomic amplification in this context. Methods. We constructed a tissue microarray (TMA) containing 915 tissue cores from 107 hormone-naïve PC and from 101 castration-resistant (CR) PC. In addition, we included 56 specimens from distant metastases. We analyzed the TMPRSS2-ERG translocation status by fluorescence in-situ hybridization and the expression profiles of ERG, AR and the proliferation marker Ki67 by immunohistochemistry. Results. Nearly half of the analyzed PC tissue specimens (untreated: 38%, castration-resistant: 46%) harbored a TMPRSS2-ERG gene fusion. Untreated PC with positive translocation status showed increased tumor cell proliferation (p<0.05). As expected, TMPRSS2-ERG gene fusion was strongly associated with increased ERG protein expression in untreated, as well as in castration-resistant PC (both p<0.0001). However, we detected a subgroup of CR PCs with the gene fusion, but without detectable ERG protein expression. This subgroup showed significantly lower levels of AR protein expression and of the androgen regulated serum PSA (both p<0.05). Conclusions. This comprehensive study comparing the TMPRSS2-ERG gene and ERG protein expression status in a large cohort of hormone-naïve and CR PC, identifies a subgroup of translocated CR PCs without ERG protein expression. Our results suggest that this subgroup may represent CR PCs with a dispensed AR pathway. Further analyses may help to explain if these tumors represent a distinct subgroup of AR independent CR PCs.

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 4562. doi:1538-7445.AM2012-4562
Abstract 4681: Inhibition of poly (ADP-ribose) polymerase-1 (PARP-1) as a strategy for targeted therapy in Ewing's sarcoma
Authors: Brenner J. Chad , Feng Felix Y. , Han Sumin , Bou-Maroun Laura M. , Patel Sonam , Goyal Siddharth V. , Liu Meilan , Lonigro Robert , Prensner John R. , Tomlins Scott A. , Chinnaiyan Arul M.
Published: August 2012

The Ewing's sarcoma family of tumors (ESFTs) are aggressive malignancies which frequently harbor EWS-FLI1 or EWS-ERG genomic fusions. Here, we demonstrate that these fusion products interact with, and depend on, poly (ADP-ribose) polymerase 1 (PARP1), a DNA damage response protein and transcriptional co-regulator. This interaction occurs in a DNA-independent manner. ESFT cell lines and xenografts are preferentially sensitive to PARP1 inhibition, and the addition of a PARP1 inhibitor to the second-line chemotherapeutic agent temozolamide resulted in complete response of all treated tumors in an EWS-FLI1 ESFT xenograft model. PARP inhibition blocked Ewing's cell line, but not control osteosarcoma or rhabdomyosarcoma cell line invasion, as well as the formation of lung metastasis in a xenograft model of ESFT. Mechanistically, the EWS-FLI1 and EWS-ERG fusions induce DNA damage, which is potentiated by PARP1 inhibition in ESFT cell lines. In a positive feedback loop, EWS-FLI1 fusions maintain the expression of PARP1 by direct regulation of the PARP1 promoter. This regulation is independently supported by a gene expression array data set of 20 ESFT patient tumors (Spearman r = 0.8165). Finally, because front-line therapy for this disease includes an intense regimen of five cytotoxic chemotherapy agents from which patients can quickly relapse with even more aggressive disease, we studied the effect of PARP inhibition in two Ewing's sarcoma cell line models derived after patient relapse. Importantly, these two cell lines maintained a preferential sensitivity to PARP1 inhibition despite their general chemoresistance. These findings suggest that targeting the EWS-FLI1: PARP1 interaction axis is a promising therapeutic strategy for the Ewing's sarcoma family of tumors.

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 4681. doi:1538-7445.AM2012-4681
Abstract 4692: ABT-888 synergizes treatment of colon cancer cell lines with irinotecan
Authors: Davidson David , Wang Yunzhe , Aloyz Raquel , Panasci Lawrence
Published: August 2012

Numerous studies have associated poly [ADP-ribosolose] polymerase 1 (PARP1) with DNA-damage repair. PARP1 rapidly localizes to DNA damage sites where it alters target proteins, (e. g. histones, topoisomerase I and DNA dependent protein kinase (DNA-PK)), through the addition of poly-ADP ribose side chains. By this mechanism, PARP1 plays a role in diverse repair pathways including homologous recombination/non-homologous end joining (HRR/NHEJ). Presently, third generation PARP inhibitors are in use clinically. These drugs have resulted in meaningful clinical responses and an increase in survival in metastatic breast and ovarian cancer patients bearing BRCA-deficient or triple negative tumors. The Abbott lead compound, ABT-888, is a potent PARP1 inhibitor that sensitizes many cancer cells in-vitro and in-vivo to temozolomide. In the present work, we hypothesized that colon cancers would be sensitized to the DNA damaging chemotherapeutic agent, irinotecan by ABT-888. Since PARP1 is involved in DNA repair, we propose that its inhibition would increase irinotecan-induced DNA damage and cell death in colon cancer cells lines. Using the sulforhodamine B assay (SRB), significant synergy (I<1 indicates synergy) was observed between ABT-888 and irinotecan at concentrations as low as 0.06 μM (Table 1). Levels of synergy observed in the SRB assay correlated with levels of PARP1 inhibition measured biochemically in cell lysates. Furthermore, 24h post treatment combinations of ABT-888/SN38 resulted in increased G2/M cell cycle arrest and increased levels of DNA damage as determined by γH2AX staining and comet assay. Additionally, 48h post treatment increased levels of apoptosis were associated with this drug combination. In conclusion this study suggests that ABT-888 may be a clinically effective adjuvant to current colon cancer therapies that include the use of irinotecan.

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 4692. doi:1538-7445.AM2012-4692
Abstract 4703: SAHA downregulation of HuR expression is responsible for reduction of cyclin D1 mRNA stability and cell transformation by EGF in Cl41 cells
Authors: Zhang Jingjie , Ouyang Weiming , Li Jingxia , Yu Yonghui , Li Xuejun , Huang Chuanshu
Published: August 2012

Previous studies have reported that anti-cancer activity of suberoylanilide hydroxamic acid (SAHA) is associated with its downregulation of cyclin D1 protein expression at transcription and translation levels. In current studies, we demonstrated that SAHA downregulation of cyclin D1 expression mainly via modulation of cyclin D1 mRNA stability in EGF-induced mouse epidermal Cl41 cell transformation (anchorage-independent colony formation) model. We found that SAHA treatment resulted in dramatic inhibition of EGF-induced cell transformation and cyclin D1 expression. Further studies showed that SAHA downregulation of cyclin D1 expression was only observed in endogenous cyclin D1, but not in reconstitutional cyclin D1 in same cells, excluding the possibility of SAHA regulation of cyclin D1 expression at levels of protein degradation. Moreover, we found that SAHA was able to inhibit EGF-induced cyclin D1 mRNA expression, whereas it did not show any inhibitory effect on cyclin D1 promoter-deriven luciferase reporter activity under same experimental conditions, suggesting that SAHA might downregulate cyclin D1 mRNA stability. This notion was greatly supported by the results showing that co-treatment of cells with SAHA and actinomycin D increased the cyclin D1 mRNA degradation in comparison to that in cells treated with actinomycin D alone. Consistent with downregulation of cyclin D1 mRNA stability, SAHA treatment also attenuated HuR protein expression, which is a well-characterized cyclin D1 mRNA-binding protein. Taken together, our results demonstrated that SAHA treatment resulted in downregulation of cyclin D1 expression by inhibition of HuR expression, which led to reduction of cyclin D1 mRNA stability and protein expression, and further in turn caused the inhibition of EGF-induced cell transformation in Cl41 cells.

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 4703. doi:1538-7445.AM2012-4703
Abstract 4716: The antitumor effect of HDAC inhibitor Tenovin-6 for gastric cancer cells
Authors: Ueno Takunori , Hirai Sachiko , Hirose Mitsuaki , Endo Shinji , Suzuki Hideo , Yamato Kenji , Hyodo Ichinosuke
Published: August 2012

Aim: Tenovin-6 is one of the small molecule inhibitors of Histone Deacetylase (HDAC), which have been to shown to suppress cancer cell growth and angiogenesis. In Japan, the mortality of gastric cancer is still high and innovative strategies are required for patients with advanced unresectable gastric cancer. We conducted this study to verify anti-tumor effect of Tenovin-6 as a single agent for gastric cancer cells and also in combination with cytotoxic agents including 5-fluorouracil (5-FU) and cisplatin (CDDP) in vitro. Moreover, we validated the effectiveness of Tenovin-6 on animal xenografted model.Methods: 4 gastric cancer cell lines, MKN-45, NUGC-4 (p53 wild), NUGC-3 (p53 mutant) and Kato-III (p53 null), were incubated with Tenovin-6 in dose and time dependent manner. Cell viability was calculated by WST-8 assay after treatment with Tenovin-6 alone or in combination with 5-FU or CDDP. Protein expression of Sirt-1, p53, p21, DR5 and Caspase-3 were monitored by western blotting to reveal mechanism of antitumor effects by Tenovin-6. In vivo xenograft model established with human gastric cancer cell line MKN-45 was treated with intraperitoneal administration of Tenovin-6 at dose of 50 mg/kg/day for 2 weeks. Results: All 4 gastric cancer cells decreased dramatically by addition of Tenovin-6 with 2.3 to 4.3 μM of IC50. The decreasing trend was not different among cell lines. The additive and synergic effects were shown with combination of 5-FU or CDDP. The expression of Caspase-3, p21 and DR5 were increased by Tenovin-6 in time dependent manner. In contrast, the expression of Sirt-1 and p53 was not changed in all periods. Also in vivo, single agent therapy with Tenovin-6 significantly reduced subcutaneous xenograft tumor growth in nude mice. Conclusion: Tenovin-6 showed excellent anti-tumor effects in all gastric cancer cells via induction of apoptosis and cell cycle arrest. It may be promising therapeutic material for gastric 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 4716. doi:1538-7445.AM2012-4716
Abstract 4724: Anti-tumor effects of SC-2001, a novel STAT3 inhibitor, in breast cancer cells
Authors: Tseng Ling-Ming , Liu Chun-Yu , Kao Yuan-Ping , Shiau Chung-Wai , Su Jung-Chen , Chang Kung-Chi , Chen Kuen-Feng
Published: August 2012

Background: STAT3 has been shown to play an important role in breast carcinogenesis and is often overexpressed in primary breast tumors. STAT3 has emerged as a novel potential anti-cancer target. Previously, we have designed SC-2001 ((Z)-2-((3-methoxy-2H-pyrrol-2-ylidene)methyl)-1H-pyrrole) and reported its efficacy in HCC cells (AACR 2011, abstract#4516). SC-2001 is structurally related to the Mcl-1 inhibitor obatoclax, and showed better antitumor effects than obatoclax in HCC cells. Here, we examined the efficacy and drug mechanism of SC-2001 in breast cancer cells. Methods: breast cancer cell lines were used for in vitro studies. Apoptosis was examined by both flow cytometry and Western blot. Signal transduction pathways in cells were assessed by Western Blot. Tyrosine Phosphatase Assay Kit was used for SHP-1 activity assay. Gene silencing was done by small interference RNA (siRNA). Results: SC-2001 inhibited tumor cell growth (by MTT assay) and induced apoptosis in several breast cancer cell lines (MDA MB-468, MDA MB-231, MDA MB-453, MCF-7). SC-2001 inhibited the protein-protein interactions between Mcl-1 and Bak similar to obatoclax. In addition, SC-2001 downregulated Mcl-1 expression by reducing its transcription, whereas obatoclax had no such effect. SC-2001 downregulated the phosphorylation of STAT3 (Tyr 705) and subsequently inhibited its transcriptional activities in a dose-dependent manner. Moreover, STAT3-regulated proteins, including Mcl-1, survivin and cyclin D1, were also repressed by SC-2001. Over-expression of STAT3 in MDA MB-468 cells protected cells from SC-2001-induced apoptosis. Importantly, SC-2001 enhanced the expression of SHP-1, a negative regulator of STAT3. Inhibition of SHP-1 by either specific inhibitor or siRNA reduced the apoptotic effects of SC-2001, indicating that SHP-1 plays a key role in mediating SC-2001-induced cell death. Conclusions: our results suggest that SC-2001 induced apoptosis in breast cancer cells, and that this effect is mediated through SHP-1-dependent STAT3 inactivation. (Supported by NSC100-2325-B-002-036, NSC98-2320-B-010-005-My3 and NSC-100-2325-B-010-007, VGHTPE V100D-005-4, 101DHA0100318, and 101DHA0100230.) Keyword: obatoclax, breast cancer, STAT3, SHP-1, apoptosis

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 4724. doi:1538-7445.AM2012-4724
Abstract 4728: Fatostatin, a novel SREBP inhibitor, suppresses cell growth and induces apoptosis in prostate cancer
Authors: Li Xiangyan , Chen Yi-Ting , Chung Leland W.K. , Huang Wen-Chin
Published: August 2012

Sterol regulatory element-binding proteins (SREBPs), SREBP-1 and SREBP-2, are transcription factors mainly controlling lipogenesis and cholesterogenesis via the regulation of genes related to fatty acid and cholesterol biosynthesis. We previously showed that SREBP-1 was highly elevated in clinical prostate cancer specimens compared with non-tumor prostate tissues. Also, SREBP-1 promoted cell viability and progression through alterations of the metabolic and signaling networks involving lipogenesis, androgen receptor (AR) and oxidative stress in prostate cancer cells. In the present study, we evaluated the therapeutic benefit of targeting the aberrant SREBP signaling in prostate cancer by using a novel SREBP inhibitor, fatostatin. Fatostatin significantly inhibited cell proliferation and suppressed anchorage-independent colony formation in soft agar in LNCaP and C4-2B prostate cancer cells. Fatostatin also reduced the abilities of in vitro migration and invasion in both cells. The results of flow cytometric analyses with annexin V/propidium iodide staining showed that fatostatin caused G2/M arrest and induced cell death. Additionally, apoptosis was observed by inducing caspase-3 and poly (ADP-ribose) polymerase cleavages and increasing caspase activities in fatostatin-treated prostate cancer cells. In the animal study, fatostatin significantly inhibited C4-2B xenograft tumor growth and markedly decreased serum PSA level compared to control group in immune-compromised mice. The results of fatostatin action in prostate cancer cells in vitro and in vivo are due to specific interruption of the proteolytic process of mature SREBP-1, -2 and their regulated gene expressions including HMG-CoA reductase, fatty acid synthase and further inhibition of lipid biosynthesis and AR signaling network. In summary, our study demonstrates for the first time that fatostatin will be developed as a novel anti-prostate cancer drug and provides new insights into cancer therapy by targeting SREBP.

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 4728. doi:1538-7445.AM2012-4728
Abstract 4750: Overcoming DDP resistance using folic acid-heparin-DDP nanoparticles
Authors: Peng Xianghong , Wang Yiqing , Chen Zuo (Georgia) , Nie Shuming , Shin Dong M.
Published: August 2012

Cisplatin (DDP) has been widely used in the treatment of different cancers. However, resistance to DDP limits its clinical application. Since reduced intracellular DDP accumulation is one of the main causes for DDP resistance, increasing the accumulation of DDP may overcome drug resistance. The folate receptor (FR) is overexpressed on the surface of many human tumors including breast, ovarian and lung cancer, but is found at low levels in normal tissues. In this study, folic acid (FA) was conjugated to heparin-DDP (FHDDP) nanoparticles to target FR. In vitro cellular uptake experiments showed that the accumulation of FHDDP nanoparticles in FR-expressing DDP-resistant KB-CP.5 cancer cells (selected from KB3-1 cells, and showing a 36-fold increase in resistance to DDP compared with KB3-1) was significantly higher than that of free DDP (9.31±2.2 ng Pt/106 cells vs 0.98±0.25 ng Pt/106 cells, Pt:0.5ug/ml, 4 hrs at 37°C). Further, the level of platinum DNA-adducts in the resistant cells treated with FHDDP nanoparticles was about 4-fold greater than in cells treated with free DDP (24.21±5.32 pg Pt/ug DNA vs 97.9±22.5 pg Pt/ug DNA). Nude mice bearing KB-CP.5 cell tumors receiving FHDDP nanoparticles (2.5mg/kg, tail vein injection, twice weekly, 5 times) showed enhanced antitumor effects as compared with mice treated with free DDP (tumor volume: 823±221mm3 vs 1450±392mm3, 21 days after treatment), while non-targeted heparin-DDP nanoparticles did not show significant difference compared with free DDP. Our results suggest that FHDDP nanoparticles have the potential to be used for overcoming DDP resistance.(This research was supported by the National Cancer Institute award 5 P50 CA128613)

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 4750. doi:1538-7445.AM2012-4750
Abstract 4764: Targeting formin autoinhibition to construct cell infrastructure
Authors: Lash-VanWhye Leanne , Kitchen-Goosen Susan , Alberts Arthur S.
Published: August 2012

Rho small GTP-binding proteins locally manage Diaphanous-related formins as they construct the cytoskeletal infrastructure that supports essential changes in cell morphology. Rho binding unlatches formin intramolecular autoinhibition. The autoinhibition is mediated by the Dia-inhibitory and -autoregulatory domains (DID & DAD) which flank the formin homology-2 (FH2) domain. The free FH2 domain guides the assembly of the linear actin and microtubule structures required for adhesion, migration, division, and polarity establishment. Microtubule stabilization is already known to be an effective anti-cancer strategy. Combined with the knowledge that formins can act as haploinsufficient tumor suppressor genes, a screen was conducted for agents that could impair autoinhibition in order to activate formins for therapeutic benefit. We have focused on two of the primary “hits” for development. VARI01 and VARI02 are nearly identical thioamide inhibitors of DID-DAD binding (IC50 values ≤ 140 nM). Exposure of cultured tumor cells to either compound triggered the expected changes in cell structure and signaling that were consistent with formin activation. These included microtubule stabilization, F-actin assembly, the activation of the transcription factor SRF, cell-cycle arrest, and eventual apoptosis. SRF senses changes in actin dynamics to activate the expression of genes that reinforce cell structure and monitor genome integrity. VARI01 and VARI02 both exhibit anti-tumor activity in xenograft studies. Ongoing studies are aimed at identifying VARI01/VARI02-sensitive tumor types and to define other potential therapeutic applications. Significance: Microtubule stabilization by taxol has proven to be an effective anti-cancer strategy, but it has significant deleterious side effects. Small-molecule formin activation serve as a novel class of anti-cancer agents that may provide tools for unlocking the biological roles of formins in signalling and disease.

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 4764. doi:1538-7445.AM2012-4764
Abstract 4770: Delivery of doxorubicin to multi-drug resistant murine xenografts via drug-loaded micelles formed from mixtures of amphiphilic triblock copolymers
Authors: Dulken Benjamin , Kim Tae Hee , Mount Christopher , Du Mengyuan , Gombotz Wayne , Pun Suzie
Published: August 2012

BACKGROUND: The toxicity of chemotherapeutic drugs is often the limiting factor in patient dosing and can be the source of severe discomfort and suffering for patients undergoing chemotherapy. For example, the anthracycline antibiotic doxorubicin (DOX) is a potent chemotherapeutic but prolonged use can also lead to permanent heart damage. Tumors can also develop resistance to DOX by increased expression of drug efflux pumps. Nanoparticle formulations of DOX have been reported to improve tumor-specific delivery via the enhanced permeability and retention (EPR) effect. METHODS: In this work, we have encapsulated DOX in polymeric micelles formed from mixtures of amphiphilic triblock copolymers. The copolymers are composed of a hydrophobic polymer segment flanked by two hydrophilic blocks. Two different copolymers were used to form the mixed micelles: a copolymer of poly(ethylene oxide) and poly(hydroxybutyrate) (PEO-PHB-PEO), and a copolymer of poly(ethylene oxide) and poly(propylene oxide) (Pluronic F127). We hypothesize that forming mixed micelles will improve drug loading and enable preparation at room temperature (where Pluronic F127 micelles are highly unstable), while facilitating drug delivery by utilizing the stability of Pluronic F127 micelles at physiological temperatures. RESULTS: We have demonstrated that the micelles formed from the mixtures of PEO-PHB-PEO and Pluronic F127 triblock copolymers provide a combination of high drug loading content and drug delivery capabilities. DOX loading for mixed micelles was greater than that for Pluronic F127-only micelles (p<0.05 for all mixture ratios). DOX encapsulated in mixed micelles demonstrated increased uptake in a drug resistant MDA-435-MDR cell line over free doxorubicin (p<.05 for all mixture ratios), while cytotoxicity of the micelle treatment was approximately equivalent to that of the free drug. Tumor treatment efficacy was assessed in mice with subcutaneous dual xenografts of MDA-435 and MDA-435-MDR (drug resistant) cell lines. Tumor suppression of drug resistant tumors was similar for encapsulated and free drug, but mice treated with micellar DOX showed no significant weight loss after treatment, while mice treated with unencapsulated DOX experienced severe weight loss after one week when compared to initial time point (18.2% +/− 2.1% weight loss, p = .0009). CONCLUSION: The cytotoxicity, drug uptake, and tumor suppression properties demonstrated by PEO-PHB-PEO/Pluronic F127 mixed micelles supports their viability as chemotherapeutic drug delivery systems for drug resistant tumors.

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 4770. doi:1538-7445.AM2012-4770
Abstract 4779: K-983, an orally available small molecule inhibitor of FGFR, demonstrated prolonged survival in a mouse orthotopic gastric cancer model
Authors: Furuta Takayuki
Published: August 2012

The receptor tyrosine kinase FGFR and its ligand fibroblast growth factor (FGF) play important roles in cell growth, migration, and survival. Dysregulation of the FGFR/FGF signaling pathway has been found in many human malignancies. K-983 was identified as a novel, potent, and orally active small molecule inhibitor of FGFR. The anti-tumor activity of K-983 was evaluated in a rat xenograft models. K-983 displayed dose dependent anti-tumor efficacy. K-983 demonstrated dose dependent inhibition of FGFR phosphorylation and prolonged survival in a mouse peritoneal dissemination model of gastric cancer. Furthermore, K-983 was examined using an orthotopic model of human scirrhous gastric cancer in nude mouse. K-983 (80 mg/kg) was orally administered once daily to the mice for 28 days. Histopathological examination revealed that K-983 inhibited tumor growth at inoculated site, invasion in gastric wall and metastasis to other organs. K-983 also significantly prolonged the survival of mice in this orthotopic tumor model. In summary, these results demonstrated the promising therapeutic potential of K-983 in anticancer 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 4779. doi:1538-7445.AM2012-4779
Abstract 4782: Two-dimensional gel analysis of protein expression profile in patients with invasive squamous cell carcinoma of the uterine cervix for identification of differentially expressed proteins in relation to response to radiation therapy
Authors: Srinivasan Radhika , Sood Swati , Ghosh Sujata , Patel Firuza
Published: August 2012

Background: Cancer of uterine cervix is the second most common cancer and an important cause of cancer related mortality among women worldwide. These patients are treated by chemoradiation, response to which is a major factor affecting their outcome. Objective: To identify differentially expressed proteins in the tissue biopsies of patients with invasive cervical cancer by two-dimensional gel electrophoresis [2-DE] and correlate it with their clinical outcome. Methods: After an informed consent, 10 patients of invasive squamous cervical cancer, stage IIB, between 40-50 years treated with chemoradiation as per standard protocol were recruited. Based on the 24 month-outcome, there were two groups (N=5): i) Evidence of disease (ED) and ii) No evidence of disease (NED). Pooled protein from the tissue biopsies were subjected to 2-DE using pH 3-10 IPG strips of 7cm length, silver stained and expression profile compared using Image Master 7.0 software (GE Healthsciences). The differentially expressed protein spots were identified and isoelectric point (pI) & Molecular Weight (MW) was determined from the plot of relative electrophoretic mobility (Rf) values of the marker against their respective molecular weight and the plot of pI marker against their respective distance from the anode respectively. The TagIdent software was used for their identification. Results: Out of a total of 327 spots identified, 89 spots showed differential expression, of which 40 were highly significant (ANOVA p <0.03) between ED vs NED groups. Conclusion: 2-DE has provided preliminaryinformation regarding differentially expressed proteins which may help identify biomarkers for prediction of outcome in patients with invasive cervical cancer which needs further confirmation. Differentially expressed proteins in NED vs ED groups in invasive cervical 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 4782. doi:1538-7445.AM2012-4782
Abstract 4786: Biomarkers for lung adenocarcinoma in salivary proteome
Authors: Villagomez Jose Guillermo , Castellanos Lyssia , Ortega Alette , Zavala Diego , Ponciano Guadalupe , Arrieta Oscar G. , Reyes Juan Pablo , Mercado Gabriela E.
Published: August 2012

Lung cancer is the major cause of cancer-related deaths and has a poor prognosis with a 5-yr overall survival of <10%, mostly due to lack of early stage diagnosis. Cigarette smoking is estimated to be responsible for >80% of the cases. Previous studies have demonstrated that former smokers have a higher risk of lung cancer compared to nonsmokers. Early detection of lung cancer would improve the overall survival of this disease. However, there is no validated screening test for lung cancer. To develop an early detection test, we selected saliva as the biofluid of choice due to its accessibility, protein content that provides diagnostic information on a variety of diseases, including cancer. To identify differentially expressed proteins in saliva from individuals with lung adenocarcinoma, whole saliva samples were collected from 6 patients with lung adenocarcinoma and 6 matched healthy controls (male individuals, >40 years, smokers >10 pack year). Proteins and peptides from whole saliva samples were analyzed by two-dimensional gel electrophoresis identifying 20 differentially expressed protein spots. Mass spectrometry analysis identified 63 peptides and revealed fourteen non-redundant proteins in saliva obtained from lung adenocarcinoma patients. Four of these proteins have been previously reported as serum biomarkers of cancer, and two of them were associated with non-small cell lung cancer. These potential biomarkers are being tested in an independent and larger cohort for validation. Patient-based saliva proteomics is a promising approach to searching for cancer biomarkers. Further characterization of these markers may provide the basis for new, noninvasive tests for screening, detection, and monitoring of high-risk individuals.

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 4786. doi:1538-7445.AM2012-4786
Abstract 4863: Ubiquitination of the breast cancer oncogene IKKα is essential for transformation
Authors: Zhou Alicia Y. , Shen Rhine R. , Kim Eejung , Chen Zhijian , Hahn William C.
Published: August 2012

IKBKE (IKKα) is a breast cancer oncogene that promotes human mammary cell transformation through the activation of the NF-κB pathway. Here we show that IKKα is modified by ubiquitination in the context of cell transformation. By mass spectrometry, we identified three lysine residues of IKKα that are ubiquitinated - Lysine30, Lysine401, and Lysine416. Mutation of all three lysine residues together eliminates IKKα ubiquitination. Single mutations of the three lysines revealed that K30 and K401 are essential for IKKα-induced malignant transformation. Although wildtype IKKα drives transformation in immortalized human cells, the IKKα K30R and IKKα K401R mutants fail to transform immortalized human cells. In addition, the IKKα K30R and IKKα K401R mutants fail to activate the NF-κB pathway and are unable to phosphorylate CYLD, a downstream target of IKKα. The ubiquitination of IKKα does not affect the protein stability of IKKα protein. Using an inducible ubiquitin-knockdown U2OS cell system, we show that IKKα specifically undergoes K63-linked polyubiquitination, a proteasome- and degradation-independent form of ubiquitination. Taken together, these data provide evidence that IKKα is subject to direct regulation by K63-linked polyubiquitination and that this ubiquitination is essential for IKKα-mediated oncogenic transformation.

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 4863. doi:1538-7445.AM2012-4863