bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2586: Pomegranate components modulate distinct pathways in prostate cancer cells
Authors: Munagala Radha , Aqil Farrukh , Gupta Ramesh C. , Higashi Richard M. , Vadhanam Manicka V.
Published: August 2012

Pomegranate juice, when provided as whole juice, modulates specific pathways in prostate cancer cells that are not reflected when its major constituents were tested independently. In order to identify these bioactive components of pomegranate, the peel (P) and arils (J) were isolated, juiced separately and extracted with 80% methanol (PE and JE). The components were enriched using a SPE C18 cartridge and, eluted as fraction I (P-I and J-I - contains mostly polar compounds) and fraction II (P-II and J-II - contains mostly non-polar compounds). Analysis by HPLC-PDA and by direct infusion analysis of these fractions by FTICR-MS revealed the J-I consisted of mostly phenolics including the three anthocyanins in its mono- and di-glycoside forms. P-I consisted mostly of punicalin and, the punicalagins A and B, while P-II and J-II consisted mostly of ellagitannins and free ellagic acid. The ORAC values as Trolox equivalent for the various fractions at 10 µg/ml were: Total peel extract (PE) - 1.64 compared with 2.74 for P-I and 4.01 for P-II, while for the total juice extract (JE) it was 2.62 compared with 7.01 for J-I and 5.47 for J-II. To identify the molecular mechanisms, the individual fractions of peel and juice were tested in prostate cancer cell lines, LNCaP and PC3 and analyzed by Cancer/Apoptosis Phospho Antibody Array and western blot analysis. In LNCaP cell line, BCL-2, PTEN, Cdc25A, p53 and STAT6 were hyper-phosphorylated by J-II, while they were unaltered by J-I. Similarly, GSK3 alpha and beta were hyper-phosphorylated by J-I and unmodified by J-II. When peel fractions were tested, Cdc25C, BCL-2, CREB and JAK were hypo-phosphorylated by P-II, whereas they were unaltered by P-I. Other molecules, such as IRS-1, MAP Kinase, PTEN, STAT3 and JAK2 were hypo-phosphorylated by P-II, and were hyper-phosphorylated by P-I. In PC3 cell line, BCL-2, p53 and CREB were hypo-phosphorylated by J-II, while they were unaltered by J-I. However, PTEN and p70 were hypo-phosphorylated by J-II and hyper-phosphorylated by J-I, while, CDK2, STATs 1, 3 and 6 were hypo-phosphorylated by P-II, they were unaltered by P-I fraction. Thus the components of peel and juice fractions demonstrate unique properties and the ability to modulate distinct pathways in androgen-dependent and independent prostate cancer cells providing a rich set of potential therapy or adjuvant applications. This work is supported by NCI grant CA143676.

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 2586. doi:1538-7445.AM2012-2586
Abstract 259: Antitumor activities of yuanhuadine, a daphnane diterpene from Daphne genkwa, via epidermal growth factor receptor signaling in non-small cell lung cancer cells
Authors: Hong Ji-Young , Park Hyen Joo , Chung Hwa-Jin , Kim Gi Dae , Song Jayoung , Lee Min Ai , Bae Song Yi , Pyee Yuna , Shin Yoonho , Lee Sang Kook
Published: August 2012

The growth inhibition and antitumor activities of yuanhuadine (1), a daphnane diterpenoid from the flowers of Daphne genkwa, were investigated in human lung cancer cells. Compound 1 exhibited a relatively selective growth inhibition against human lung cancer cells compared to other solid human cancer cell lines. The potent antiproliferative activity by 1 was associated with cell-cycle arrest and modulation of cell signaling pathways. Cell-cycle arrest in the G0/G1 and G2/M phase was induced by 1 in A549 human non-small cell lung cancer cells, and these events were correlated with the expression of checkpoint proteins including the up-regulation of p21, and down-regulation of cyclins, cyclin-dependent kinases 2 (CDK2) and 4 (CDK4), and c-Myc. Compound 1 also suppressed the expression of the Akt/mammalian target of rapamycin (mTOR) and its downstream effector molecules including p70 S6 kinase (p70S6K) and eukaryotic initiation factor 4E-binding protein 1 (4EBP1). The ligand-induced epidermal growth factor receptor (EGFR) and c-Met signalings were also inhibited by 1. The oral administration of 1 (0.5 mg/kg body weight, daily) for 14 days inhibited tumor growth in athymic xenograft nude mouse model bearing human lung A549 cells significantly, without any overt toxicity. Synergistic antiproliferative effects of compound 1 were also found in combination with the EGFR inhibitor gefitinib. Cell-cycle arrest and suppression of Akt/mTOR and EGFR signaling pathways might be plausible mechanisms of actions for the antiproliferative and antitumor activity of 1 in human non-small cell lung cancer 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 259. doi:1538-7445.AM2012-259
Abstract 2652: Randomized phase III clinical trial of a combined treatment with carnitine + celecoxib +/− megestrol acetate for patients with cancer-related anorexia/cachexia syndrome
Authors: Madeddu Clelia , Dessi Mariele , Panzone Filomena , Serpe Roberto , Antoni Giorgia , Recchia Francesco , Mantovani Giovanni
Published: August 2012

Aims: A phase III, randomized non-inferiority study was carried out to compare a two-drug combination (including nutraceuticals, i.e. antioxidants) with carnitine + celecoxib +/− megestrol acetate for the treatment of cancer-related anorexia/cachexia syndrome (CACS): the primary endpoints were increase of lean body mass (LBM) and improvement of total daily physical activity. Secondary endpoint was: increase of physical performance tested by grip strength and six minute walk test. Methods: Sixty eligible patients were randomly assigned to: arm 1, L-carnitine 4 g/day + Celecoxib 300 mg/day or arm 2, L-carnitine 4 g/day + celecoxib 300 mg/day + megestrol acetate 320 mg/day, all orally. All patients received as basic treatment polyphenols 300 mg/day, lipoic acid 300 mg/day, carbocysteine 2.7 g/day, Vitamin E, A, C. Treatment duration was 4 months. Planned sample size was 60 patients. Results: The results did not show a significant difference between treatment arms in both primary and secondary endpoints. Analysis of changes from baseline showed that LBM (by dual-energy X-ray absorptiometry and by L3 computed tomography) increased significantly in both arms as well as physical performance assessed by 6MWT. Toxicity was quite negligible and comparable between arms. Conclusions: The results of the present study showed a non-inferiority of arm 1 (two drug-combination) vs arm 2 (two drug-combination + megestrol acetate). Therefore, this simple, feasible, effective, safe, low cost with favourable cost-benefit profile, two-drug approach could be suggested in the clinical practice to implement CACS treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2652. doi:1538-7445.AM2012-2652
Abstract 2653: Racial and ethnic differences in survival after cancer diagnosis
Authors: Lovy Andrew J. , Gorlick Richard G. , Geller David S.
Published: August 2012

Introduction: Racial differences in cancer survival have been well established based on national and independent cancer registries. Multiple extrinsic factors including stage, socioeconomic status (SES) and treatment have been postulated as causative, questioning whether race is a significant independent predictor of cancer survival. The purpose of this study was to evaluate racial differences in overall cancer survival rates in a diverse NY population. Methods: Study population included 18,556 patients (28% White, 31% African American (AA), 27% Hispanic) diagnosed and/or treated for primary cancer from 1/1/2000 through 1/1/2007 at Montefiore Medical Center (MMC), identified through MMC's Cancer Registry. Demographics, clinical information and survival were obtained through the Cancer Registry. SES was determined using census tract data. Kaplan-Meier survival curves and Cox proportional hazards regression were used to compare overall survival by race. Results: White patients were older (67.6 vs. 60.9 & 58.7) and of higher SES than AA and Hispanic patients, with similar rates of stage III/IV cancer and surgical treatment. Unadjusted analysis revealed that Whites had a significantly higher risk of death compared to all other patients (HR 1.29, 95%CI 1.23-1.35). Adjusting for age, sex, stage at diagnosis, surgical treatment and SES in all cancer types combined, revealed a significantly higher risk of death compared to all other patients (HR 1.14, 95%CI 1.06-1.21). However, race was no longer a significant predictor of cancer survival in multivariate models stratified by cancer type. Conclusion: Contrary to published literature, White patients had significantly lower survival rates than AA and Hispanic patients in unadjusted models. Racial differences in cancer survival diminished in adjusted models, and were absent when stratifying by cancer type. These results suggest that racial disparities in cancer survival may be better explained by variables such as age, stage at presentation, SES, treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2653. doi:1538-7445.AM2012-2653
Abstract 2663: Phase I study of azacitidine and cisplatin in patients with advanced head and neck or non-small cell lung cancer
Authors: Liao Yu-Min , Mirshahidi Hamid , Zhang Kangling , Mirshahidi Saied , Williamson Stephen , Hsueh Chung-Tsen
Published: August 2012

Background: DNA hypomethylation agents such as azacitidine may be used for epigenetic sensitization of cisplatin treatment, and therefore leading to improvement of treatment outcome. We conducted a phase I study of azacitidine and cisplatin to determine safety, toxicity, and effect of global DNA methylation in peripheral blood mononuclear cells. Methods: Azacitidine was given as subcutaneous injection daily from day 1 to day 5, and cisplatin was given at 75 mg/m2 IV on day 8. The treatment was repeated every 28 days. Four patients received azacitidine 37 mg/m2 daily from day 1 to day 5 on dose level 1. Two patients were treated on dose level 2, and received azacitidine 60 mg/m2 daily from day 1 to day 5. Peripheral blood samples were collected before and after azacitidine treatment. DNA was extracted from peripheral blood mononuclear cells, and subjected to gas chromatography/mass spectrometry to measure the level of global DNA methylation, as reflected by the ratio of 5-methylcytosine to the sum of 5-methylcytosine and cytosine. Results: No patient had dose-limiting toxicities. Four patients completed at least 2 cycles of treatment, and were evaluated for clinical response by CT scan and DNA methylation effect. One partial response was noted in patient with metastatic tongue cancer after 2 cycles of treatment on dose level 1 of azacitidine. This patient received total 3 cycles of treatment and achieved free of progression for about 15 months on chemotherapy holiday. One patient with recurrent salivary gland cancer achieved stable disease after 2 cycles of treatment on dose level 2, and maintained in free of progression for more than 6 months after 4 cycles of treatment. There was inhibition of global DNA methylation after azacitidine treatment ranging from 12 to 36% in all 4 patients. Furthermore, the inhibition of global DNA methylation was associated with the increase of fetal hemoglobin in patients’ peripheral blood. Conclusion: Sequential administration of azacitidine followed by cisplatin is well tolerated, and demonstrates encouraging activity in patients with advanced head and neck cancer. This combination deserves further clinical investigation to study the correlation between clinical response and biological effects.

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 2663. doi:1538-7445.AM2012-2663
Abstract 2666: Receptor protein tyrosine phosphatase type kappa is a prognostic marker, essential for regulating malignant behavior of glioma cells
Authors: Agarwal Supreet , Sibenaller Zita , Ryken Timothy C. , Assem Mahfoud
Published: August 2012

Malignant gliomas have a poor survival resulting from rapid growth of the tumor and extensive dispersion throughout the brain parenchyma. Discovery of tumor-specific biomarkers hold great promise in improving survival and thus call for better understanding of regulatory signals driving glioma cell migration and dispersal. Using genome-wide exploration, we have identified frequent genetic alterations at the Receptor protein tyrosine phosphatase type kappa (PTPRK) locus. These PTPRK alterations were among the top significant and very relevant to glioma biology since PTPRK is a dephosphorylation regulator of tyrosine phosphorylation (TP) signaling, a frequently altered signaling cascade in glioma as reported previously. Understanding PTPRK function will provide relevant novel insights into the molecular mechanisms underlying glioma cell growth and migration. PTPRK expression was analyzed by quantitative-RT-PCR and western blot. Mutations of PTPRK were identified by sequencing full length PTPRK transcripts from a subset of patients’ samples showing PTPRK loss of heterozygosity. Two major PTPRK variants were cloned and expressed in cells harboring altered PTPRK (U87MG cell line) to study their effect on PTPRK phosphatase activity and functionality. PTPRK locus undergoes allelic loss in glioma and contributes to cancer phenotype. It is an independent prognostic marker in glioma. Patients with altered PTPRK (23%) have shorter survival compared to those with normal PTPRK molecular status (p=0.001, Log-rank test, 14 vs. 30 months). PTPRK mRNA as well as protein levels were lower in tumor biopsies compared to non-tumor specimens. Mutational analysis revealed missense mutations in the phosphatase domain, resulting in altered PTPRK phosphatase activity, possibly due to changes in active site conformation. Further, PTPRK alterations changed expression profiles of the PTPRK protein as a result of altered post-translational processing. Reconstitution of PTPRK expression in glioma cells (U87MG cell line) harboring PTPRK deletions suppressed cell growth and migration which was alleviated by the PTPRK variants to differing extent. PTPRK knockdown via shRNA down-regulated its onco-suppressive function compared to PTPRK expressing cells. Our findings provide the first evidence of the key role of PTPRK expression in glioma and shed light on the biological consequences of mutations in the PTPRK gene which significantly alter PTPRK functionality. Biological relevance of PTPRK variants will be determined and validated on a large scale.

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 2666. doi:1538-7445.AM2012-2666
Abstract 2676: A secreted tissue engineered extracellular matrix (shECM) prevents recurrence in post-resection pancreatic cancer models
Authors: Pinney Emmett , Menen Rhiana , Hassanein Mohamed , Montes-Camacho Mayra , Naughton Gail , Bouvet Michael , Hoffman Robert M.
Published: August 2012

Pancreatic cancer is a highly treatment-resistant cancer. Gemcitabine, which is first-line therapy, results in toxicity without meaningful survival benefit. We report here that a extracellular matrix (ECM) secreted by human embryone fibroblasts in culture inhibit human pancreatic cancer growth and post-surgical recurrence in an orthotopic nude mouse models. Post-resection, all untreated control mice had recurrent disease. In contrast, only 2 of 8 mice (p=0.004) in the treatment group had palpable masses after 6 weekly intravenous treatments. In a second study, targeting the one surgical site with ECM treatment followed by 8 weekly i.p. treatments of ECM, 7 of 8 treated mice survived beyond 10 weeks while none of the untreated control mice survival 8 weeks. No measurable toxicity was observed in the treatment group. These results demonstrate the potential of ECM treatment of pancreatic 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 2676. doi:1538-7445.AM2012-2676
Abstract 2678: Traditional Chinese Medicine (TCM) in combination with S. typhimurium A1-R for cancer treatment
Authors: Zhang Yong , Zhang Nan , Su Shibing , Hoffman Robert M. , Zhao Ming
Published: August 2012

Traditional Chinese Medicine (TCM) has long shown promise for the treatment of cancer. We have previously developed a bacterial cancer therapy strategy using the genetically-engineered strain, Salmonella typhimurium A1-R, which is auxotrophic for arg and leu, which attenuates growth in normal tissue but allows high tumor virulence. S. typhimurium A1-R is effective against metastatic human prostate, breast, and pancreatic cancer cell lines as well as osteosarcoma, fibrosarcoma, and glioma cell lines in clinically-relevant nude-mouse and syngeneic-mouse models. The current study evaluated TCM as combination treatment during bacterial therapy in metastatic mouse models of human cancer. TCM prevented hemorrhagic spots on the skin, liver, and spleen after bacterial infection. Fur discoloration and reduced activity was also observed in mice treated with S. typhimurium A1-R. These symptoms were ameliorated when bacterial treatment was combined with TCM. In a syngeneic lung cancer metastatic model in immunocompetent mice, the combination of TCM and S. typhimurium A1-R enabled S. typhimurium to be safely administered at a dose of 2×105 CFU twice a week i.v. in immunocompotent mice with increased treatment efficacy compared to monotherapy with S. typhimurium A1-R. The results of the present study suggest that combination of S. typhimurium A1-R and TCM has important potential for cancer 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 2678. doi:1538-7445.AM2012-2678
Abstract 2679: Color-coded selective chemotherapy of methioninase-synchronized S-phase cancer cells expressing cell-cycle-specific fluorescent reporters: A general approach to the treatment of cancer
Authors: Yano Shuya , Li Shukuan , Han Qinghong , Tan Yuying , Fujiwara Toshiyoshi , Hoffman Robert M.
Published: August 2012

Cancer cells of all types have a generally elevated requirement for methionine compared to normal cells. This phenomenon is termed methionine-dependence and may be due to excessive methylation reactions in cancer cells, since methionine is the global source of cellular methyl groups. Deprivation of methionine α,β lyase (methioninase or METase) selectively arrests cancer cells during late S-phase, where they are highly sensitive to chemotherapy drugs which damage DNA. Cancer cells, transformed to express different color fluorescent reporters during specific phases of the cell cycle (fluorescent ubiquitination-based cell cycle indicator [FUCCI]), were used to monitor the onset of the S-phase block due to methionine deprivation effected by METase. The S-phase-blocked cancer cells fluoresced yellow or green, in contrast to cancer cells in G1 which fluoresced red. Cancer cells, synchronously blocked in S-phase by METase and identified by their yellow-green fluorescence, were administered chemotherapy drugs which interact with DNA or block DNA synthesis such as doxorubicin, cisplatin or 5-fluorouracil. We have termed this strategy color-coded chemotherapy. The procedure was highly effective against the cancer cells. In contrast, treatment of cancer cells with drugs only, and without methioninase-effected S-phase synchrony, led to the majority of the cancer cell population being blocked in G phase (red fluorescent) where they were resistant to the drugs. The selective anticancer strategy and technology described in this report, whereby cancer cells are selectively and synchronously blocked in S-phase in the most drug-sensitive phase of the cell cycle where they are identified by fluorescent reporters and then treated with S-phase-specific-drugs, should be a general approach to the treatment of cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2679. doi:1538-7445.AM2012-2679
Abstract 2685: Imaging the various mechanisms of cancer-cell killing by Salmonella typhimurium A1-R
Authors: Zhao Ming , Zhang Yong , Uchugonova Aisada , Salz Renee , Liu Fang , Suetsugu Atsushi , Zhang Lei , Hoffman Robert M.
Published: August 2012

We have developed a genetically-modified bacterial strain of Salmonella typhimurium, auxotrophic for leucine and arginine, which also expresses green fluorescent protein (GFP), termed S. typhimurium A1-R. S. typhimurium A1-R was found to be effective against metastatic human prostate, breast, and pancreatic cancer cell lines as well as osteosarcoma, fibrosarcoma and glioma cell lines in clinically-relevant nude mouse models. To understand the tumor-killing mechanism of S. typhimurium A1-R-GFP, we have studied their interaction with fluorescence-labeled cancer cells in vitro and in vivo. S. typhimurium-GFP invasion, proliferation, and virulence in the cancer cells were visualized by confocal fluorescence microscopy with the Olympus FV1000. We found that A1-R-induced cancer cell death had different mechanisms in different cancer cell lines. Some S. typhimurium A1-R-treated cancer cell lines became apoptotic and necrotic, and other cell lines burst after extensive intracellular bacterial growth. The time for S. typhimurium A1-R-GFP to kill the majority of the cancer cells varied from line to line, ranging from 2 hours to 72 hours. The understanding of the various mechanisms of cell killing by S. typhimurium A1-R will be important for its use as a general therapeutic for 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 2685. doi:1538-7445.AM2012-2685
Abstract 2689: Inhibition of metastasis of circulating tumor cells by extracellular matrix secreted by human embryonic cells
Authors: Menen Rhiana , Pinney Emmett , Hassanein Mohamed K. , Bouvet Michael , Hoffman Robert M.
Published: August 2012

We have previously demonstrated the increased metastatic potential of human prostate cancer PC-3 CTCs compared to their parental counterparts in both chick embryo and mouse models. In the present study, we tested the effects of human embryonic extracellular matrix (ECM), secreted by human embryonic cells including conditioned medium (CM) and a semi-solid form (hECM), on metastasis of PC-3 CTC in the chick embryo model. The chorioallentoic membrane (CAM) of 18 chicken embryos were inoculated with either 106 PC-3 human prostate cancer cells or PC-3 CTCs, both stably expressing GFP. Twelve hours later, embryos were divided into 6 groups each containing three embryos: PC-3 parental control; PC-3 + CM; PC-3 + hECM; CTC control; CTC + CM; and CTC+ hECM. Twelve hours following inoculation of the cells, a single dose of 100 μL of either treatment was given to the appropriate group. Embryo brains were removed on day 8 post-inoculations, and processed for cryo-sectioning, generating 3 slides of brain tissue per embryo taken at various depths. Imaging was performed using the IV-100 scanning laser microscope (Olympus Corp, Tokyo, Japan) in order to count metastatic foci. PC-3 controls had an average of 11.1 metastatic foci compared to 2.55 in the PC-3 + hECM group (p< 0.0001) and 2.76 in the PC-3 + CM group (p< 0.0001) The treatment showed even greater response on the CTC cells which an average of 30.9 metastatic foci in the CTC controls compared to 4.38 in the CTC + hECM group (p< 0.0001) and 4.18 in the CTC + CM group (p< 0.0001). Thus, human embryonic secreted ECM compound drastically decreased the metastatic potential of human prostate cancer CTCs in the chick embryo model.

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 2689. doi:1538-7445.AM2012-2689
Abstract 2693: Pharmacogenetic determinants associated with sunitinib-induced toxicity and ethnic difference in Korean patients with metastatic renal cell carcinoma
Authors: Kim Hye Ryun
Published: August 2012

Purpose: The aim of this study was to investigate the pharmacogenetic determinants of sunitinib-related toxicity and ethnic difference in metastatic renal cell carcinoma (mRCC) among Korean patients. Experimental Design: A pharmacogenetic study was performed in 65 patients with mRCC treated with sunitinib (50mg orally once daily for 4 weeks-on/2 weeks-off). Detailed data regarding the toxicity of sunitinib, including thrombocytopenia, neutropenia, anemia, and hand-foot syndrome (HFS) were prospectively collected in a clinical trial program (n=38) or general oncology practice (n=27) using sunitinib. A total of 12 genetic polymorphisms in 7 candidate genes (CYP1A1, ABCG2, ABCB1, PDGFRα, VEGFR2, RET, and FLT3) were analyzed for an association with treatment-related toxicity using Pearson-χ2 test. Results: Common grade 3 or 4 treatment-related toxicities were thrombocytopenia (36.9%), neutropenia (18.4%), anemia (7.7%), and HFS (12.3%). Patients carrying an ABCG2 421 A/A genotype developed significantly more grade 3 or 4 thrombocytopenia, neutropenia, and HFS: odds ratio [OR] 10.15; P=0.03, thrombocytopenia, OR 17.33; P=0.003, neutropenia, OR 33.60; and P=0.005, HFS. Conclusion: c.421C>A genetic polymorphism in the ABCG2 gene may be most associated with the risk of sunitinib-related toxicity in mRCC patients. Considering the high frequency of ABCG2 c.421C>A SNP in Asian individuals, this may lead to differential toxicities among ethnic groups.

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 2693. doi:1538-7445.AM2012-2693
Abstract 2729: Suppression of hematogenous metastasis by mouse and murine/human chimeric anti-Aggrus/podoplanin antibodies
Authors: Takagi Satoshi , Fujita Naoya
Published: August 2012

Tumor cell-induced platelet aggregation has been known to facilitate hematogenous metastasis by increasing the arrest of tumor cell emboli in the microvasculature. We have previously established metastatic variants of cancer cells and identified Aggrus (also known as podoplanin, T1alpha, gp36) as a platelet aggregation-inducing factor expressed on the surface of highly metastatic variants. Aggrus, a type-I transmembrane sialomucin-like glycoprotein, induced platelet activation via interaction with one of its counter receptor CLEC-2 (C-type lectin-like receptor 2) expressed on platelet surface. Thus, Aggrus-induced platelet aggregation did not require any plasma components. Moreover, Aggrus has been shown to be up-regulated in many different types of cancers and to be associated with poor outcome. Thus, Aggrus must be a useful cancer specific antigen in addition to a useful therapeutic target for developing hematogenous metastasis inhibitors. In this study, we established an anti-human Aggrus monoclonal antibody, designated as P2-0, and investigated its potential as a hematogenous metastasis inhibitor. We observed that P2-0 antibody could inhibit Aggrus-CLEC-2 interaction detected by in vitro competition assay and could suppress Aggrus-induced platelet aggregation in a concentration-dependent manner. Moreover, P2-0 antibody decreased the number of Aggrus-mediated pulmonary metastatic foci in in vivo experimental mouse model. We then cloned complementarity-determining region of P2-0 antibody and generated murine/human chimeric P2-0 antibody. We revealed that chimeric P2-0 antibody retained Aggrus-neutralizing activity and inhibited the experimental pulmonary metastasis of cancer cells that endogenously expressed Aggrus on their surface. These data indicate that P2-0 antibody and its chimeric antibody are useful as inhibitors of Aggrus-mediated hematogenous metastasis and expected to be applied for the clinical examination of Aggrus-targeted cancer therapy. This study was supported in part by a grant from the Program for the Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation, Japan.

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 2729. doi:1538-7445.AM2012-2729
Abstract 273: Transcriptional regulation of L1CAM in endometrial cancer
Authors: Schirmer Uwe
Published: August 2012

The L1 cell adhesion molecule (L1CAM, CD171) was initially described as a cell surface protein in the developing nervous system involved in neural outgrowth and axon guidance. During the last years it became evident that high expression levels of L1CAM are observed in many human carcinomas and are linked to tumor progression. L1CAM expression in tumors can augment cell motility and invasive potential and lead to constitutive NFκB activation. We showed before that in endometrial cancer (EC) the expression of L1CAM was correlated with poor prognosis. ECs can be subdivided in two types; a less aggressive type I and a highly aggressive type II. We showed that L1CAM expression was absent in type I ECs, whereas all cases of type II tumors (n=272) were L1CAM positive. We also provided evidence that in L1CAM-negative EC cell lines L1CAM expression can be upregulated by TGF-b1 treatment involving the transcription factor SLUG. In the present study, we have investigated additional mechanisms that can affect L1CAM expression. These mechanisms involve the modification of histone proteins and their packaging to the DNA as well as the direct modification of the DNA by DNA-methyltransferases (DNMTs). We show that the treatment of EC cell lines with Histon-deacetylase-Inhibitors (HDACIs) and/or DNMT inhibitors lead to increased L1CAM expression at the mRNA and protein level. Furthermore, we observed that the methylation pattern of the L1CAM-promoter differed between L1CAM positive and negative cell lines. The RE-1 silencing transcription factor (REST) complex seems to be responsible for the silencing of the L1CAM expression in non-neuronal tissues. The REST complex is associated with chromatin-remodeling complexes, the recruitment of HDACs as well as DNMTs and thereby leads to the silencing of the respective promoter element. We observed that the knockdown of REST in L1CAM negative EC cell lines leads to a dramatic increase in L1CAM expression. Likewise the overexpression of REST in L1CAM positive EC cell lines diminished L1CAM expression. We conclude that the loss or reduced expression of REST in concert with epigenetic elements could represent an additional and novel mechanism to cause aberrant L1CAM expression in ECs.

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 273. doi:1538-7445.AM2012-273
Abstract 2735: Effects of metformin on a non-small cell lung cancer cell line with an EGFR mutation
Authors: Kitazono Satoru , Takiguchi Yuichi , Ashinuma Hironori , Kitazono Miyako , Kitamura Atsushi , Tada Yuji , Kurosu Katushi , Sakaida Emiko , Sekine Ikuo , Tatsumi Koichiro
Published: August 2012

Background:Metformin, one of the most widely prescribed oral agents for treating diabetes mellitus, has been speculated to have antineoplastic effects. The effects, however, have not been fully understood. Experimental Design:A human non-small-cell lung cancer cell line, PC9 (with an EGFR mutation of the exon 19 E746-A750 deletion), was used to evaluate antineoplastic effects of metformin in vitro and in vivo. For in vitro experiments, the cells were exposed to metformin, gefitinib or cisplatin to observe inhibitory effects on cell proliferation. At 24, 48 and 72 hours after the start of the culture with the agents, cell cycle analysis, measurements of caspase 3, 8 and 9 activities, and evaluation with Hoechst staining and APOPercentage TM assay, respectively, were performed. For in vivo experiments in SCID mouse with a xenograft subcutaneous tumor, gefitinib (150 mg/kg, oral gavage) or metformin (250 mg/kg, i.p.) was daily administered to the mice. Results:Gefitinib at a concentration of 0.03 μM, cisplatin of 1.5 μg/ml and metformin of 10 mM exerted equivalent inhibitory effects on cell proliferation in vitro. At these particular concentrations, metformin did not induce significant apoptosis over control as assessed by Hoechst staining, APOPercent assay and caspase 3 activity, whereas gefitinib and cisplatin did induce apoptosis dramatically. Metformin failed to alter cell cycle distribution, whereas gefitinib and cisplatin caused G1/G0 an S phase accumulation, respectively. Metformin did not inhibit growth of already grown subcutaneous tumors, whereas the tumors shrank dramatically with gefitinib treatment. After almost complete disappearance of the tumors by gefitinib, however, they regrew by gefitinib withdrawal. Metformin did significantly suppress the regrowth of the tumors after gefitinib withdrawal. Conclusions: Metformin has inhibitory effects on growth of a human non-small cell lung cancer cell line with a sensitive EGFR mutation in vitro and in vivo, and its mechanism may not depend on apoptosis or cell cycle alteration. Although the agent does not suppress growth of already grown tumors, it significantly suppresses tumor regrowth after gefitinib withdrawal.

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 2735. doi:1538-7445.AM2012-2735
Abstract 2737: Anti-proliferative action of metformin on various types of human lung cancer cell lines
Authors: Ashinuma Hironori , Takiguchi Yuichi , Kitazono Satoru , Kitazono-Saitoh Miyako , Kitamura Atsushi , Tada Yuji , Kurosu Katsushi , Sakaida Emiko , Sekine Ikuo , Tanabe Nobuhiro , Tatsumi Koichiro
Published: August 2012

[Background] Metformin has a potential to inhibit tumor growth in various cancer cell lines in vitro and in vivo. [Purpose] To elucidate anti-proliferative actions of the agent on various types of human lung cancer cell lines, and to evaluate combined effects with cisplatin. [Methods] Cell lines used consisted of four human lung cancer cell lines, RERF-LC-AI (squamous cell carcinoma), A549 (adenocarcinoma), IA-5 (large cell carcinoma) and WA-hT (small cell carcinoma). Colonogenic assay was employed to evaluate colonogenicity with exposure to metformin for 1, 24 hours and 10 days. In cell surviving assay, 100,000 cells per plate were cultured for 4 days with metformin and/or cisplatin, and survived cells were counted. [Results] Metformin inhibited colonogenecity in every cell line in a dose-dependent manner when cells were exposed to the agent for 10 days, whereas 1- and 24-hour exposure yielded not significant or only mild effects, with the degree varied depending on cell lines. With cell surviving assay at the metformin concentrations where 70% (IC70) and 30% (IC30) of the cells survived for each cell line, apoptosis was not increased over control groups in every cell line except for WA-hT that showed significant increase in apoptosis with metformin. Significant alteration in cell cycle distribution with the agent was not observed in all of the 4 cell lines. For evaluating combination treatment consisting of metformin and cisplatin, cisplatin at doses of IC50 and IC10 was combined with metformin. Inhibitory effects of metformin on cell proliferation were slightly suppressed with cisplatin at its dose of IC50 in all except for in A549. High dose (IC10) of cisplatin almost completely countervailed or even reversed the metformin effects in all cell lines, again except for A549 where a modest but significant sub-additive effect was observed. [Conclusions] Metformin has anti-proliferative effects on various types of human lung cancer cell lines. Although apoptosis was observed only in a small-cell lung cancer cell line, its effects may not depend on apoptosis in other 3 cell lines. Alteration of cell cycle distribution was not observed in all cell lines. Metformin and cisplatin were antagonistic especially with higher dose cisplatin.

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 2737. doi:1538-7445.AM2012-2737
Abstract 2753: The addition of low dose 17-DMAG to BIBW-2992 effectively inhibits lung cancer cells with acquired resistance by T790M mutation
Authors: Rho Jin Kyung , Choi Yun Jung , Kim Sun Ye , Choi Chang-Min , Lee Jae Cheol
Published: August 2012

The majority of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) is caused by secondary T790M mutation. We established gefitinib or erlotinib-resistant cells which acquired different proportions of T790M from PC-9 cells with activating EGFR mutation. Anti-cancer effects of EGFR modulation and strategies overcoming resistance were investigated. The amount of T790M and EGFR gene copy number were proportional to increase of resistance. BIBW-2992, an irreversible EGFR-TKI, could suppress the growth and survival of cells harboring lower level of T790M while it failed in cells with higher level of T790M although p-EGFR was completely down-regulated in both, suggesting blocking of phosphorylation of EGFR is not sufficient to inhibit these cells. However, EGFR siRNA treatment could inhibit cell growth regardless of T790M level and resistance showing that EGFR dependency would be persistent. 17-DMAG, a HSP90 inhibitor, also effectively controlled both resistant cells. Considering a report showing that a HSP90 inhibitor was not clinically effective possibly because of inadequate dose, the efficacy of low dose 17-DMAG was examined. Interestingly, we found that combined treatment of BIBW-2992 with low dose 17-DMAG was very effective even in cells with higher level of T790M although either of both showed no activity in single treatment suggesting it would be one of promising strategies with immediate clinical implications overcoming T790M-mediated resistance. Further investigations on exact mechanism should be pursued even though capability of HSP90 inhibitor to modulate NF-kB signaling can be one of possible explanations.

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 2753. doi:1538-7445.AM2012-2753
Abstract 2754: Cinnamoyl quinolines are novel autophagy inhibitors and enhance antitumor activity of 2-deoxyglucose
Authors: Yih Ling-Huei , Kakadiya Rajesh , Wu Yi-Chen , Kuo Hsiao-Hui , Su Tsann-Long
Published: August 2012

Cinnamides exhibit a variety of biological properties including antineoplastic, antiallergic, and anti-infective activities etc. Modifications of the amide function of cinnamides by linking with aliphatic or heterocyclic moieties results in the formation of cinnamoyl derivatives with anticancer activities. In the present study, we prepared a series of cinnamoyl quinolines and evaluated their antitumor activity in a panel of cancer cell lines. These derivatives reduced cancer cell viability by inducing apoptosis and showed little to no cross-resistance to vinblastine- or cisplatin-induced multidrug resistance. Interestingly, the cinnamoyl-quinoline-6-yl derivatives (BO-2110 and BO-2112) induced cytoplasmic vacuolization and the processing and recruitment of microtubule-associated protein 1 light chain 3 (LC3) to autophagosomes. However, these agents caused the accumulation and aggregation of p62/SQSTM1. Co-treatment of cinnamoyl-quinoline-6-yl derivatives with bafilomycin A1 did not induced a further increase in p62/SQSTM1 accumulation. These results indicated that cinnamoyl-quinoline-6-yl derivatives blocked autophagy. Since autophagy enables cancer cells to temporarily avoid cell death when nutrient deprivation or exposing to chemotherapeutic agents, cinnamoyl quinolines may sensitize cancer cells to death and augment chemotherapy-induced apoptosis. Co-treatment of these derivatives with 2-deoxyglucose (2DG, glycolysis inhibitor), but not cisplatin (DNA alkylating agent), camptothecin (Topo I inhibitor), doxorubicin or etoposide (Topo II inhibitor), significantly enhanced cell death in the tested tumor cell lines. Collectively, these results suggest that BO-2110 and BO-2112 are effective autophagy inhibitors and promising candidates that may be used in combination with 2DG for treating cancers.

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 2754. doi:1538-7445.AM2012-2754
Abstract 2769: The development of 19-substituted benzoquinone ansamycins as potential anticancer drugs
Authors: Chang Chuan-Hsin , Kitson Russell , Siegel David , Moody Christopher J. , Ross David
Published: August 2012

Benzoquinone ansamycins (BQA's) were the first class of Hsp90 inhibitors developed but the use of the prototype agent geldanamycin was limited by hepatotoxicity. One major mechanism of toxicity of the BQA class is manifested via the electrophilic properties of the quinone and alkylation of cellular nucleophiles at the 19-position. We developed novel 19-substituted BQA's in the geldanamycin (GA), 17-AAG and 17-DMAG series as a means to prevent arylation of cellular nucleophiles. 19-substituted BQA's inhibited recombinant Hsp90 and did not react with thiols at the 19-position while marked thiol reactivity could be detected using their parent quinones. We examined the effects of different substitution patterns at the 19-position on the ability of BQAs to inhibit Hsp90 and induce growth inhibitory effects in breast cancer cell lines. We found that both 19-phenyl and 19-methyl BQA's in the 17-AAG, 17-DMAG and GA series were effective inhibitors of purified Hsp90 as demonstrated using an ATPase assay particularly in the presence of NQO1 to generate the hydroquinone ansamycin. Only 19-phenyl substituted BQAs were effective growth inhibitory agents in breast cancer cell lines using an MTT assay while 19-methyl substituted BQAs in the GA, 17-AAG and 17-DMAG series demonstrated only modest growth inhibition. The growth inhibitory effects of 19-phenyl BQAs were potentiated in cells containing high NQO1. Notably, 19-phenyl DMAG had similar growth inhibitory and apoptotic effects as its parent quinone, DMAG. These data demonstrate that the 19-phenyl BQAs had marked growth inhibitory, apoptotic, and client protein expression effect in breast cancer cells compared to 19-methyl BQAs (Supported by CA 51210).

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 2769. doi:1538-7445.AM2012-2769
Abstract 2771: HSP90 inhibitor NVP-AUY922 exhibits the potent antiproliferative effect in gastrointestinal cancer cells with a lack of PTEN expression
Authors: Takaoka Munenori , Bao Xiaohong , Hao Huifang , Fukazawa Takuya , Yamatsuji Tomoki , Tomono Yasuko , Sakurama Kazufumi , Naomoto Yoshio
Published: August 2012

Background: The heat shock protein 90 (HSP90), a molecular chaperone, has provoked great interest as a promising molecular target for cancer treatment, since it is involved in regulating the conformation, stability and functions of key oncogenic proteins. At present, a variety of chemical compounds targeting HSP90 have been developed and have shown convincing anti-neoplastic activity in various preclinical tumor models. Several inhibitors of HSP90 have been reported exhibiting more antitumor efficacy in PTEN-null tumor xenograft. The aim of our study was to evaluate the antitumor effects of a novel HSP90 inhibitor, NVP-AUY922, in human malignant cells, especially in PTEN-null cancer cells. Materials and Methods: Human esophageal cancer (TE-1, TE-4, TE-8, TE-10), gastric cancer (NUGC-3, NUGC-3/IL), pancreatic cancer (PANC-1, MIAPACA-2, Bxpc3) and mutant c-kit-driven lymphocytes (Ba/F3-KE11, Ba/F3-K642, Ba/F3-820Mz), mimicking gastrointestinal stromal tumor cells, were examined. WST assay and/or trypan blue exclusion assay were conducted to assess the anti-proliferation effects of NVP-AUY922 in those cells mentioned above. The activity of cell growth-related molecules such as AKT, ERK, and PTEN were investigated by western blot analysis. Genetic modification using siRNA and expression vector were applied for silencing and overexpression of PTEN, respectively, in cancer cells. Results: Cell viability assays showed that NVP-AUY922 potently inhibits the proliferation of human cancer cell lines, regardless of 5-FU resistance or c-Kit mutation, by which the sensitivity to imatinib can be affected. PTEN-null TE-4 cells exhibited more sensitivity to NVP-AUY922. Interestingly, The sensitivity to NVP-AUY922 was increased by silencing PTEN expression in intact PTEN-expressing TE-10 cells and decreased by exogenous transduction of PTEN in TE-4 cells. Furthermore, NVP-AUY922 significantly inhibited the activity of AKT and ERK in TE-4 cells, but not in TE-10 cells. Conclusion: Inhibition of Hsp90 function by NVP-AUY922 exhibited strong cytotoxicity against most of the cancer cells, including some anticancer drug-resistant cells, revealing its potential for a novel therapeutic alternative to cancer treatment. In addition, this study suggests that the expression status of PTEN determines the sensitivity to HSP90 inhibitor for cancer treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2771. doi:1538-7445.AM2012-2771