bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 730: Evaluation of micro-RNA biomarker panel on biopsies and non-invasive samples in nasopharyngeal carcinoma
Authors: ying ying Szeto Carol , KWONG Dora Lai-wan , Lee Anne , Wai-tong Ng , LEE Victor Ho-fun , Lung Maria Li
Published: August 2012

Nasopharyngeal carcinoma (NPC) is a prevalent malignant disease in Southeast Asia among the Chinese population. Being stable, sensitive and dysregulated in cancer, microRNAs (miRNAs) are potential biomarkers for NPC. Several screening and mechanistic studies have identified potential miRNA biomarkers from different systems, however, integrative biomarker panels including both human and EBV-encoded miRNAs are still lacking. Herein we evaluate the usefulness of a potential biomarker panel on biopsies and non-invasive samples (nasal swab and plasma) in NPC. By optimizing methodology, we can detect the expression of miRNAs in different samples (biopsies, nasal swabs, and plasma) from minimal amounts of samples. We have identified eight miRNA candidates in the literature as a testing panel and analyzed their expression levels in 20 paired tumor/normal biopsies samples. Five (EBV-mir-BART2, -BART4, -BART22, -BART7, and has-mir-34c) show significant differences in miRNA expression levels. Unbiased hierarchical clustering analysis of all eight candidates yield clusters of cases may be related to different causes and treatment outcomes of NPC. Five out of the eight miRNAs were chosen for further screening on a total of 40 paired biopsy samples to evaluate the significance of each cluster. Four (EBV-mir-BART2,-BART22, -BART7, and has-mir-34c) show significant expression (p<0.05) in paired tumor/normal biopsies samples. The miRNA expression in nasal swab samples with paired biopsies was also analyzed. Expression in cell-free plasma RNAs of the miRNA candidate panel has also been examined to evaluate the usefulness of this panel as non-invasive biomarkers. Our results suggested the feasibility of utilizing a miRNA biomarker panel for diagnosis/prognosis of NPC using biopsies and of nasal swabs and plasma to minimize the invasiveness and amounts of samples needed for diagnosis. Acknowledgement: This work was supported by University Grants Council, HKSAR Area of Excellence grant AoE/M-06/08 to MLL.

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 730. doi:1538-7445.AM2012-730
Abstract 735: Metaplastic columnar lined esophagus without goblet cells show phenotypic similarities to Barrett's Esophagus
Authors: Srivastava Supriya , Liew Mei Shan , McKeon Frank , Xian Wa , Ho Khek Yu , Teh Ming
Published: August 2012

Aim: Barrett's esophagus (BE) is a premalignant condition, predisposing to esophageal adenocarcinoma (EAC). However, some EAC arise in columnar lined esophageal mucosa devoid of goblet cells. Therefore, the purpose of this study was to evaluate the biological properties of non-goblet columnar lined esophagus (CLE) and elucidate the relationship between BE and high grade dysplasia of esophagus (HGD). Material and Methods: Endoscopic biopsies from esophagus of 47 patients with columnar metaplasia with goblet cells (BE; n=28), columnar metaplasia with non-goblet cell (CLE; n=10), BE associated high grade dysplasia (HGD; n=9) and cardia with intestinal metaplasia (CIM; n=5) were included in the study. These groups were immunostained for, Villin (intestinal marker), CK7 (columnar epithelium marker), Claudin 3 (epithelial tight-junction marker) and MUC4 (membranous mucin). Statistical analysis between different groups was performed using the paired or unpaired t- test using SPSS v 15.0. A p value <0.05 was considered significant. Results: Patients with BE, CLE and HGD showed positivity for Villin, Claudin 3 and CK7 in 79%, 93%, 54% in BE group; 60%, 100% and 80% in CLE group and 78%, 100% and 34% in HGD group respectively. In metaplastic epithelium (BE and CLE) there was no statistically significant difference in expression of these markers (p>0.05). Patients with CLE without goblet cells showed immunopositive results for these markers, statistically similar to those of HGD group and also revealed significantly higher values compared to those of controls (cardia with intestinal metaplasia). MUC4 was expressed higher in 96% of BE cases as compared to 67% of HGD cases (mean, 81.85±27.74 vs 48.89±45.12). CLE without goblet cells did not express MUC4. Conclusion: Our results indicate that metaplastic columnar lined epithelium without goblet cells suggest biological properties of intestinal differentiation. It also shows that apart from BE, CLE without goblet cells may also have neoplastic potential.

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 735. doi:1538-7445.AM2012-735
Abstract 737: Hypofractionated stereotactic radiotherapy to surgical cavity after resection for brain metastasis
Authors: Yu Hsiang-Hsuan M. , Tsai Ya-Yu
Published: August 2012

Recurrence at resection cavity after resection for brain metastasis is high; whole brain radiotherapy (WBRT) decreases this risk but may cause neurocognitive complication. Post-operative radiosurgery to resection cavity is a new technique to decrease recurrence and spare patients from WBRT, but single high-dose fraction may potentially increase radiation-induced necrosis. Hypofractionated stereotactic radiotherapy (SRT) delivers higher radiobiological dose which may potentially provide higher local tumor control, and fractionation may lower risk of radiation-induced injury. To investigate the efficacy of post-operative hypofractionated SRT after resection of brain metastasis, we retrospectively analyzed 52 consecutive patients with brain metastasis from solid tumors who underwent surgical resection followed by SRT to the resection cavity for at least one metastasis from April 2008 to March 2011. A total of 25 to 30 Gy in 5 daily fractions over 1-week period was prescribed to the resection cavity with a margin of 2 mm expansion (median dose 28.5 Gy) between 2 to 4 weeks post-operatively. Synchronous oligometastasis, if present, was treated with radiosurgery. No patients received upfront WBRT. All patients were followed up for at least 6 months. Follow-up was assessed using clinical evaluation and MR imaging every 2 to 3 months. The radiographic end point was local tumor control at resection cavity. Preliminary analyses showed the actuarial local control rate of 86% at 12 months at the resection site. For those with synchronous brain metastasis, the rates of local control at resection cavity and oligometastases were not statistically significantly different (p=0.4). Approximately 30% of the patients developed distant brain recurrence during follow-up, and 70% of these patients received salvage radiosurgery while the rest received WBRT. None of the patient developed radiation-induced injury at the resection site. Multivariate analysis showed that number of metastasis was associated with higher rate of distant brain recurrence. Hypofractionated SRT appears to be safe and effective in preventing recurrence at resection cavity following surgical resection of brain metastasis and may spare many patients from WBRT and its adverse 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 737. doi:1538-7445.AM2012-737
Abstract 738: Re-irradiation for recurrent malignant glioma by boron neutron capture therapy
Authors: Kawabata Shinji , Miyatake Shin-Ichi , Hiramatsu Ryo , Kuroiwa Toshihiko , Ono Koji
Published: August 2012

OBJECTIVE: Boron neutron capture therapy (BNCT) is based upon the nuclear reaction that occur when non-radioactive boron-10 (10B) is irradiated with low energy neutrons to produce high energy a particles (10B[n, alpha] 7Li). In order for BNCT to be successful, a sufficient amount of 10B and neutrons must be delivered to the tumor. We have applied tumor selective particle radiation known as boron neutron capture therapy (BNCT) to malignant brain tumors. As there has been no standard treatment for recurrent gliomas so far, it has been difficult to evaluate the results of BNCT for recurrent malignant gliomas. Here we introduce the survival benefit of BNCT for recurrent malignant glioma patients, with special reference to new RPA classification recurrent malignant gliomas advocated by New Approaches to Brain Tumor Therapy CNS Consortium (NABTT, JCO 2007). METHODS: Since 2002, we have treated 27 cases of recurrent malignant gliomas with BNCT. All cases had been treated by standard treatment including radiotherapy mainly by XRT prior to BNCT. After BNCT, patients were followed by MRI and F-BPA (amino acid)-PET. Cause of death was estimated by these modalities. Also overall survival was evaluated with special reference to RPA classes advocated by NABTT as above. RESULTS: All cases showed radiographical improvement at once. The median survival time (MST) of BNCT-treated recurrent gliomas was 10 months, while that of total NABTT cases (N=333) was 7 months. MST of NABTT RPA classes were 25.7, 17.2, 3.8, 10.4, 5.6, 6.4 and 4.9 months for classes 1 to 7, respectively. The MST of BNCT-treated cases were 33, 24, 10, 9, 9, 12 and 11, for classes 1 to 7, respectively. BNCT showed good survival benefit especially for the most poor prognostic group (RPA class 3+7) from 4.4 months (NABTT) to 9.6 months. Conclusion: BNCT could prolong the survival of patients with recurrent gliomas not only for the good prognosis group but also for the poor prognosis group. Some of the prognostic factors of malignant glioma, such as KPS, tumor location, extent of surgery, are not independent factor each other. BNCT seems to have the survival benefits for such situations due to its tumor cell selectivity.

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 738. doi:1538-7445.AM2012-738
Abstract 756: Pharmacokinetics of curcumin and curcumin O-glucuronide in healthy volunteers characterized by liquid chromatography-tandem mass spectrometry
Authors: Liu Zhongfa , Chen Wei , Fan-Havard Patty , Yee Lisa D. , Mo Xiao-Kui , Stoner Gary D. , Chan Kenneth K.
Published: August 2012

Purpose: To develop and validate a liquid chromatography-tandem mass spectrometric method (LC-MS/MS) for simultaneous quantification of curcumin and curcumin O-glucuronide (COG) in human plasma, and to characterize the pharmacokinetics (PK) of curcumin and COG in healthy volunteers after separate oral administrations of Curcumin C3 Complex (CC3C) and nano-emulsion Curcumin (NEC) using this method. Methods: The study was conducted at The Ohio State University (OSU) with the approval of the OSU Institutional Review Board. CC3C and NEC were administered orally to four healthy volunteers and plasma levels of curcumin and COG were determined by a validated LC-MS/MS method. The pharmacokinetic parameters were calculated using WinNonlin computer software. Clinical safety assessment included vital signs and biochemical measurement on the day of each PK study and three weeks following each PK study to monitor potential toxicity. A mixed effect model was used for comparing the concentration of curcumin or COG at each time point after the oral consumption of CC3C or NEC across each donor using SAS 9.2 software. Results: An LC-MS/MS method for simultaneous quantification of curcumin and COG in human plasma with the lower limit of quantification of 2 ng/mL was established. The within-day and between-day validation parameters were consistent with the Food and Drug Administration's (FDA) criteria for Good Laboratory Practice analytical methods. A cross-over pharmacokinetic study in the four healthy volunteers revealed that 2g NEC delivered much higher plasma concentrations of free curcumin (1.5-8.0 ng/mL vs. non-detectable, p<0.001, paired t-test) and COG (550-3300 ng/mL vs. 3.0-167 ng/mL, p<0.05, paired t-test) when compared to 4g of CC3C. The total drug exposure of curcumin and COG from NEC in human volunteers is 26 to 350-fold higher than that of CC3C. No adverse events were noted following administrations of CC3C and NEC. No abnormalities were noted on the hematologic, clinical chemistry, and metabolic profiles at prescreening, and at 3 weeks following CC3C and NEC administration. Conclusion: NEC represents a novel formulation of curcumin with enhanced oral absorption of curcumin and is worthy of evaluation for future clinical applications. Supported by NIH-NCI-R21 CA135478 [ZL], UL1RR025755 [NCRR] and the BioMedical Mass Spectrometry Laboratory [KC & ZL].

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 756. doi:1538-7445.AM2012-756
Abstract 764: Multiple testing in a phase II trial design for the detection of tumor subtypes
Authors: Roberts John D. , Ramakrishnan Viswanathan
Published: August 2012

Tumor subtypes may be especially responsive to a particular treatment. Phase II designs that incorporate tumor subtypes typically require prospective tumor subtyping and stratified patient enrollment. These features are cumbersome, discourage patient and investigator participation, and select for patients with relatively indolent disease who can wait for subtyping results. Previously we described a two stage phase II design that permits tumor subtyping at the end of stage 1 and that does not require stratified enrollment (Clin Cancer Res 2011;17:5538). In this design results from stage 1 are used to select an appropriate stage 2 objective. The stage 2 objective may be either to estimate the response rate of either the tumor or a subtype or a formal test of the hypothesis that the response rate for a subtype is greater than the overall response rate. Previously, although our design accommodated multiple subtypes in stage 1, it allowed further testing of only a single subtype in stage 2. Here we expand upon the previous design by describing a method to evaluate multiple tumor subtypes in stage 2 with appropriate adjustments for multiple testing. As there are practical limits on sample size, these adjustments tend to reduce power. We identify strategies to limit loss of power by taking advantage of tumor biology. For example, when tumor subtypes are mutually exclusive or nested, some comparisons can be made without further “alpha spending”. We explore the promise of our approach for the efficient identification of clinically significant tumor subtypes more responsive to a specific targeted 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 764. doi:1538-7445.AM2012-764
Abstract 767: CXL017 re-sensitizes multidrug resistant leukemia cells to chemotherapy via modulating Bcl-2 family proteins and SERCA proteins
Authors: Das Sonia , Hermanson David , Xing Chengguo
Published: August 2012

Multidrug resistance (MDR) is a major hurdle in the treatment of cancer and there is a pressing need for new therapies. We have recently developed CXL017, derived from a Bcl-2 and SERCA dual inhibitor, with selective cytotoxicity towards MDR cancer cells. In this study, we present new evidence for its therapeutic potential in treatment of MDR cancers and offer mechanistic insights. CXL017 significantly and selectively suppresses the growth of tumors derived from the MDR cancer cell line, HL60/MX2, in vivo. In addition, HL60/MX2 failed to develop stable resistance to CXL017 upon prolonged exposure. In comparison, HL60/MX2 acquired greater than 2000-fold resistance to cytarabine (Ara-C). Remarkably, instead of acquiring cross-resistance, HL60/MX2 cells exposed to CXL017 were re-sensitized to standard therapies (10 to 100-fold). Western blotting analyses revealed that HL60/MX2 has significantly elevated levels of Mcl-1, Bax and Bak relative to HL60. HL60/MX2 also over-expresses SERCA2 and SERCA3 with elevated endoplasmic reticulum (ER) calcium content. CXL017 exposure significantly down-regulates Mcl-1 and Bax but causes up-regulation of Bcl-XL, SERCA2, and SERCA3 proteins, along with a reduction in ER calcium content. Given the well-established functions of Bcl-2 family proteins and ER calcium in drug resistance, our results suggest that the reduction in Mcl-1 and the decrease in ER calcium content are responsible for CXL017 induced chemosensitization of MDR cancer cells. These results support the strategy to develop dual inhibitors of anti-apoptotic Bcl-2 proteins and SERCA to treat MDR 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 767. doi:1538-7445.AM2012-767
Abstract 771: Changes in the subcellular pharmacology of cisplatin, in response to treatment of cells with an anti-Gli1 shRNA
Authors: Fain Jason C. , Amable Lauren , Gavin Elaine , Reed Eddie
Published: August 2012

We have demonstrated that a specific anti-Gli1 shRNA can ablate Gli1 in A2780-CP70 cells, with no effect on Gli2. Upon ablation of Gli1, platinum-DNA adduct repair is inhibited, along with inhibition of ERCC1, XPD, XRCC1, and c-jun. We therefore asked the question of whether there were other subcellular pharmacologic effects, with respect to cisplatin, in these cells. We assessed total cellular accumulation of cisplatin under three conditions: pretreatment with the anti-Gli1 shRNA; pretreatment with a scrambled shRNA control; and pretreatment with Fugene which was used as the transfection agent. We also assessed platinum-DNA adduct levels, at the same time points that total cellular accumulation was measured. All platinum measurements were performed by AAS with Zeeman background correction. In each independent experiment (n=4), we plated cells on day 1; transfected with shRNA 16-18 hours later; treated with cisplatin for 1 hour at 30 μM (the IC50 under these conditions); and, harvested cells at different time points. Those time points were: zero hours (immediately after the 1 hour cisplatin treatment); 6 hours later; or 12 hours later. At zero hours, measured levels of pt-DNA damage were equivalent between the three conditions. At zero hours, measured levels of total cellular platinum were reduced by 40-45% in anti-Gli1 treated cells; suggesting inhibition of cellular accumulation. At 6 hours, scrambled controls showed reductions in total cellular platinum by ∼40%, demonstrating excretion of cisplatin. This is consistent with previously published work (JCI 1991). Cells treated with anti-Gli1 shRNA showed no reductions in total cellular platinum at 6 hrs, nor at 12 hrs; suggesting inhibition of the processes by which cells may eliminate this drug. Cells treated with anti-Gli1 shRNA showed statistically significant inhibition in platinum-DNA adduct repair, as compared to cells treated with scrambled shRNA. Control cells repaired 67.5% of pt-DNA adducts over 12 hours; as compared to 23.7% repair in cells treated with anti-Gli1 shRNA; p2=0.015. We conclude that ablation of Gli1 in cisplatin-resistant A2780-CP70 human ovarian cancer cells, has at least three effects on the subcellular pharmacology of cisplatin. There is inhibition of processes that determine: cellular uptake of drug; cellular elimination of drug; and platinum-DNA adduct repair.

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 771. doi:1538-7445.AM2012-771
Abstract 784: Synergistic effect of magnetic nanoparticles and docetaxel on prostate cancer cells in vitro
Authors: Watanabe Masatoshi , Sato Akiko , Kurioka Daisuke , Ishiguro Hitoshi , Uemura Hiroji , Kubota Yoshinobu
Published: August 2012

Current chemotherapy approaches to prostate cancer need to be improved because of limitations by lack of specificity and systemic toxicity although docetaxel-based chemotherapy has been shown to improve the quality of life in patients with the advanced disease. Nanotechnology has prompted new and improved materials for biomedical applications with particular emphasis in therapy and diagnostics. Especially, magnetic nanoparticles (Fe3O4) have potential applications in drug delivery, cancer diagnosis and therapy. We have shown that magnetic nanoparticles would modify the effect of chemotherapy in cancer cells. In this study, we analyzed the combined effects of docetaxel and magnetic nanoparticles on prostate cancer cell lines (LNCaP, DU145, and PC3) and their mechanisms. The combination of docetaxel and magnetic nanoparticles more effectively inhibited cancer cell growth and induced apoptosis. Only magnetic nanoparticles produced reactive oxygen species (ROS), however did not inhibit cancer cell growth significantly. Magnetic nanoparticles were also found to have the effect on the expression of ABC transporter genes, and have the possibility to inhibit the PI3K-Akt-mTOR pathway. These results suggest that magnetic nanoparticles may result in favorable development of current chemotherapy for the advanced disease and leading to new tumor ablation therapies.

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 784. doi:1538-7445.AM2012-784
Abstract 788: CK2α regulates MRP1 mediated multidrug drug resistance in cancer via phosphorylation of Thr249
Authors: Stolarczyk Elzbieta I. , Reiling Cassandra J. , Pickin Kerry A. , Coppage Ryan , Knecht Marc , Paumi Christian M.
Published: August 2012

We have previously shown that the function of Ycf1p, yeast ortholog of MRP1, is regulated by yeast casein kinase 2α (Cka1p) via phosphorylation at Ser251. In this study we explore whether CK2α, the human homolog of Cka1p, regulates MRP1 by phosphorylation at the semi-conserved site, Thr249. Knockdown of CK2α in MCF7-derived cells expressing MRP1 (MRP1 CK2β(−)) resulted in increased doxorubicin sensitivity. MRP1-dependent transport of LTC4 and E217αG into vesicles derived from MRP1 CK2β(−) cells was decreased compared with MRP1 vesicles. Moreover, Thr249 to alanine mutation (MRP1-T249A) also resulted in decreased MRP1-dependent transport, while a phosphomimicking mutation (MRP1-T249E) led to dramatic increase in MRP1-dependent transport. Studies in tissue culture confirmed these findings showing increased intracellular doxorubicin accumulation in MRP1 CK2β(−) and MRP1-T249A cells compared to MRP1 cells. Inhibition of CK2 kinase by DMAT resulted in increased doxorubicin accumulation in MRP1 cells, but not in MRP1 CK2β(−), MRP1-T249A or MRP1-T249E cells, suggesting that CK2α regulates MRP1 function via phosphorylation of Thr249. Indeed, CK2α and MRP1 interact physically and recombinant CK2 phosphorylates MRP1 derived peptide in vitro in Thr249 dependent manner, while knockdown of CK2α results in decreased phosphorylation at MRP1-T249. The role of CK2 in regulating MRP1 was confirmed in other cancer cell lines where CK2 inhibition decreased MRP1-mediated efflux of doxorubicin. This study supports a model in which CK2α potentiates MRP1 function via direct phosphorylation of Thr249. We believe that addition of CK2 inhibitors to cancer treatments will improve therapeutic outcome in part due to reversal of MRP1- mediated drug resistance.

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 788. doi:1538-7445.AM2012-788
Abstract 808: Endocrine resistance: The influence of stromal-epithelial interaction on intracrine pathways of oestrogen synthesis
Authors: Purohit Atul , Odendaal Joshua M. , Palmieri Carlo
Published: August 2012

Resistance to endocrine therapy, both de novo and acquired, poses a significant clinical challenge. Two primary pathways, the aromatase and steroid sulphatase (STS) pathways, exist for the production of estrogenic steroids from androstenedione and DHEAS, respectively. Numerous studies have demonstrated an upregulation of these pathways in breast tumour tissue compared to normal breast tissue. Therapies targeting oestrogenic pathways form the mainstay of the pharmacological approach to breast cancer treatment. The SERM, Tamoxifen, and aromatase inhibitors (AI) such as Letrozole, have shown substantial clinical benefit in hormone-dependent breast cancer patients. Additionally, to date one STS inhibitor has entered clinical trials. Despite this success, a proportion of hormone-receptor positive patients do not respond to hormone therapy with 40% of patients who initially show a response to Tamoxifen relapsing. As little is known about the influence of the tumour microenvironment, particularly stromal-fibroblasts, on these pathways in the development of endocrine resistance, we have examined the influence of stromal-epithelial interactions on the expression of enzymes of steroidogenesis in endocrine resistant cells. Wild type and tamoxifen-resistant MCF-7 breast cancer cells (MCF-7wt and MCF-7TamR, respectively) were indirectly co-cultured with primary fibroblasts derived from breast tumour and reduction mammoplasty samples, using 0.4μm 6-well inserts. Expression levels of 17α-hydroxysteroid dehydrogenase types 1 (17α-HSD1) and 2, (17α-HSD2), aromatase, oestrogen receptor ≤ (ERα) and β, progesterone receptor, steroid sulphatase (STS), and steroid sulphotransferase were assessed basally and in co-culture using RT-PCR. Significantly higher basal expression of STS (p<0.001), aromatase (p=0.012) and 17α-HSD2 (p=0.032) was seen in fibroblast versus epithelial lines. On co-culture a 2.5 - 5 fold up-regulation of aromatase expression was seen in MCF-7wt and MCF-7TamR (p=0.009, p=0.046 respectively). The expression of STS (p=0.150, p=0.05) and 17α-HSD1 (p=0.001, p=0.071) was also increased. Epithelial lines showed a significant down-regulation in ERα on co-culture. Both malignant and normal fibroblast lines demonstrated up-regulation of aromatase expression on co-culture (p=0.037). These data suggest that stromal-fibroblasts make a significant contribution to the tumour hormonal environment through both expression and modulation of the pathways associated with intracrine oestrogen production. The differential effect of cell-type combinations implicates these interactions in both oestrogen sensitivity and insensitivity.

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 808. doi:1538-7445.AM2012-808
Abstract 812: Identification of novel anticancer drugs as substrates of organic anion transporting polypeptide 1B3 using a cell viability assay
Authors: Obaidat-Hays Amanda L. , Hagenbuch Bruno
Published: August 2012

Organic anion transporting polypeptide 1B3 (OATP1B3) belongs to a superfamily of transport proteins that mediate the uptake of a wide range of endogenous and exogenous compounds into cells. Under normal conditions OATP1B3 is selectively expressed at the basolateral membrane of human hepatocytes. Recently, OATP1B3 expression has been detected in a variety of different cancers and because of its multispecificity OATP1B3 is thought to be a potential therapeutic target to enhance drug delivery into cancer cells. Thus far, only a few anticancer drugs have been identified as substrates of OATP1B3 including docetaxel, paclitaxel, methotrexate and imatinib. Given that OATP1B3 can potentially be used as a target for anticancer drug delivery we need to identify additional anticancer drug substrates of OATP1B3. Therefore, we aimed at establishing a cell based high throughput assay to screen the NCI approved oncology drug set for novel cytotoxic substrates of OATP1B3. Using a luminescent cell viability assay, we measured the viability of wild-type and OATP1B3-expressing Chinese Hamster Ovary (CHO) cells in the absence or presence of the anticancer drugs. After we identified positive hits we further characterized them with respect to concentration dependencies. We identified etoposide, bleomycin, oxaliplatin and plicamycin as compounds that kill OATP1B3-expressing CHO cells at much lower concentrations than wild-type CHO cells suggesting that these compounds are substrates of OATP1B3. Using secondary assays we confirmed that etoposide is a substrate of OATP1B3 and we are currently testing the remaining compounds for direct uptake by OATP1B3. In conclusion, this cell-based viability assay is a good tool to identify novel cytotoxic OATP1B3 substrates that can potentially be used to treat OATP1B3-expressing 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 812. doi:1538-7445.AM2012-812
Abstract 815: The role of spleen tyrosine kinase (SYK) in paclitaxel resistant ovarian cancer
Authors: Gaillard Stephanie , Stoeck Alexander , Davidson Ben , Wang Tian-Li , Shih Ie-Ming
Published: August 2012

The development of resistance to paclitaxel represents a general challenge in clinical oncology, particularly in ovarian serous carcinomas (OSCs), the most common and lethal form of ovarian cancer. Initial treatment with a taxane and platinum combination regimen is standard and while the majority of women will achieve a major response to this treatment, the mortality of ovarian cancer is directly related to the tumor's development of chemoresistance. Using a proteomic approach to comprehensively compare the protein expression profiles of primary and recurrent OSC clinical samples, our laboratory previously identified several novel candidate proteins associated with the development of recurrent OSC. In this study, we focus on evaluating one of these proteins, spleen tyrosine kinase (SYK), a non-receptor tyrosine kinase, and its role in the maintenance of paclitaxel resistance in OSC. We have determined that increased expression of SYK in OSC tumor cells obtained from ascites and/or pleural effusion fluid is associated with a poorer response to chemotherapy. Using a cell culture system, we found that two OSC cell lines show an increased expression of SYK after development of in vitro resistance to paclitaxel treatment. More importantly, we demonstrate that SYK small compound inhibitors significantly resensitize the tumor cells to paclitaxel. Our results suggest that SYK plays a role in maintaining paclitaxel resistance in OSC. Recently an oral SYK inhibitor was found to be safe and effective in treating rheumatoid arthritis and non-Hodgkin lymphoma; our results suggest a potential translational use for this inhibitor as a chemosensitizing agent in patients with OSC.

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 815. doi:1538-7445.AM2012-815
Abstract 826: Yes-associated protein increases hepatocellular carcinoma chemosensitivity by modulating p53
Authors: Bai Nan , Zhang Chunyan , Chang Antao , Zhang Zhuhong , Yin Jing , Xiang Rong , Liu Chenghu
Published: August 2012

The function of the yes-associated protein (YAP) transcription co-activator remains an enigma since reports indicate that YAP either acts as an oncogene or enhances apoptosis. In this study, we found that YAP expression occurs in 51% of human hepatocellular carcinoma (HCC) biopsies while phosphorylated YAP (pYAP) expression occurs in 33% of Grade II and III HCCs. In addition, we found that YAP and p53 expression/ nuclear translocation increase in chemo-treated HepG2 cells. In chemo-treated HepG2 cells transfected with WT-YAP or blank (control), the chemosensitivity of the HepG2 + WT-YAP cells increases compared to HepG2+blank cells. In chemo-treated HepG2 cells transfected with Flag-S94A (TEAD binding domain mutant), Flag-W1W2 (WW domain mutant), or blank (control), the chemosensitivity and p53 expression/ nuclear translocation of the HepG2+Flag-S94A cells and HepG2+Flag-W1W2 cells decrease compared to HepG2+blank cells. In chemo-treated HepG2 cells transfected with Flag-5SA-S94A or Flag-5SA-W1W2, the chemosensitivity and p53 expression/ nuclear translocation of the HepG2+Flag-5SA-S94A cells and HepG2+Flag-5SA-W1W2 cells increase again compared to HepG2+Flag-S94A cells or HepG2+Flag-W1W2 cells respectively. These results indicate that YAP promotes chemosensitivity by modulating p53 during chemo-treatment, and YAP-modulated p53 activity requires the TEAD binding domain and the WW domain. In summary, our data indicate that YAP functions as a tumor-suppressor that enhances apoptosis by modulating p53 activity during chemo-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 826. doi:1538-7445.AM2012-826
Abstract 841: CCL2 and interleukin-6 regulate EMT-mediated chemo-resistance in prostate cancer
Authors: Zou Chunlin , Lu Yi , Mizokami Atsushi , Keller Evan T. , Pienta Kenneth J. , Zhang Jian
Published: August 2012

Prostate cancer (PCa) is one of the most common cancers and the second leading cause of cancer-related death in western countries. In China, the incidence has increased, unexpectedly and sharply, for the past 20 years. Chemo-resistance, in various types of tumor including PCa, is one of the critical problems in cancer therapy. In PCa, for example, the resistance to taxanes (DTX) accounts for tumor relapse and progression resulting in skeletal metastasis and high mortality. Epithelial-mesenchymal transition (EMT) has been suggested as a key process that dominates chemo-resistance through unknown mechanisms. In the current study, we reported that CCL2 and interleukin-6 (IL-6) serve as key molecules in EMT-mediated chemo-resistance in PCa cells using previously established paclitaxel-resistant cells, DU145-TxR and PC-3-TxR. Parental DU145 and PC-3 cells were used as controls. We observed that DU145-TxR and PC-3-TxR cells produced significantly higher amount of CCL2 and IL-6 compared to their parental cells, determined by cytokine array. In addition, DU145-TxR and PC-3-TxR cells demonstrated EMT phenotypes including hall maker changes in E-cadherin and Vimentin expression and transcriptional factor (Snail, ZEB1, and Slug) alterations. These changes were reversed partially by adding neutralizing antibodies against either CCL2 or interleukin-6. Interestingly, synergistic effects were observed by adding both reagents. Finally DU145-TxR, PC-3-TxR, or their parental cells were implanted into SCID mice and the mice were given DTX (10mg/kg, biweekly) and tumor growth was monitored for 4 weeks. Both DU145-TxR and PC-3-TxR tumors grew faster than the parental tumors. Tumor tissues were collected and total RNAs were isolated. Levels of CCL2 expression in these tumors were significantly higher in DU145-TxR and PC-3-TxR tumors. IHC analysis confirmed corresponding staining of EMT markers such as E-cadherin and Vimentin expression. Together we concluded that CCL2 and IL-6 regulates EMT-mediated chemo-resistance in prostate cancer. These results are of high impact of human prostate cancer chemotherapy. Supported by U.S. Department of Defense PC061231 (J. Zhang), National Natural Sciences Foundation Key Project (81130046, J. Zhang) and National Natural Science Foundation Project (81171993, Y. Lu).

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 841. doi:1538-7445.AM2012-841
Abstract 843: Influx of thiosemicarbazone NSC73306 is mediated by the cisplatin and copper transporter SLC31A1 (CTR1)
Authors: Fung King Leung , Tepede Abisola K. , Pluchino Kristen M. , Hall Matthew D. , Gottesman Michael M.
Published: August 2012

Multidrug resistance (MDR) mediated via the drug efflux pump P-glycoprotein (P-gp/ABCB1) overexpression is a major obstacle to the success of chemotherapy to combat many cancers. Clinically, P-gp expression correlates with poor clinical response to chemotherapy in patients, and its presence on the blood-brain barrier (BBB) can reduce the efficacy of otherwise promising agents targeted to the brain. Attempts to reverse drug resistance by inhibiting P-gp have not been successful, and we have been investigating alternative strategies for targeting multidrug-resistant (MDR) cancer cells. One such strategy is the use of NSC73306, a thiosemicarbazone compound that is selectively toxic towards P-gp-expressing cells. Here, we investigate the cellular uptake of NSC73306, and examine its interaction with P-gp and copper transporter 1 (SLC31A1, CTR1). In porcine kidney epithelial LLC-PK1 cells, cellular accumulation of NSC73306 is saturable, and overexpression of human P-gp in LLC-PK1 cells sensitizes them to NSC73306. Accumulation of [3H]NSC73306 in LLC-PK1 cells is inhibited by cyclosporin A (IC50 = 500 μM), verapamil (IC50= 700 μM) and cisplatin (IC50 = 77 μM). Among these compounds, we tested the effect of cisplatin on NSC73306. Selective toxicity of NSC73306 to P-gp-expressing LLC-PK1 cells was reversed by cisplatin in a dose-dependent manner (up to 40 μM). In contrast, the amount of cisplatin required to kill 50% of LLC-PK1 cells was not affected by increasing the amount of NSC73306 (up to 2μM). Since cisplatin could inhibit NSC73306 uptake, we hypothesize that these two compounds share a common uptake pathway, but cisplatin has additional routes of entry into cells. Cells transiently expressing CTR1 showed significantly higher [3H]NSC73306 accumulation. In contrast, CTR1 knockdown cells had lower [3H]NSC73306 uptake. Although cisplatin is a transport substrate of CTR1, we found that CTR1 protein was overexpressed in cisplatin-resistant KB-CP20 and BEL7404-CP20 cell lines. Compared to their parental cisplatin-sensitive cells, even though these resistance cells show decreased cisplatin accumulation, suggesting that CTR1 is not rate-limited for cisplatin uptake. These cell lines also showed higher [3H]NSC73306 uptake compared to their parental cells. In contrast, MDR cell lines overexpressing P-gp had unchanged CTR1 protein levels. Incubation with NSC73306 for 24 hours down-regulated CTR1 in a dose-dependent manner. These results suggest that NSC73306 is a transport substrate of CTR1.

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 843. doi:1538-7445.AM2012-843
Abstract 855: The antileukemia drug nilotinib inhibits invasion and metastasis of colorectal cancer cells by targeting the receptor tyrosine kinase DDR1
Authors: Leroy Cédric , Tosti Priscillia , Simon Valérie , Robert Bruno , Sirvent Audrey , Roche Serge
Published: August 2012

Colorectal cancer (CRC) is one of the leading causes of cancer-related death in the Western world. Cancer that metastasizes to distant sites is usually not curable, although chemotherapy can extend survival. Recently, a novel immunotherapy approach based on targeting of the tyrosine kinase (TK) EGFR was shown to significantly improve the survival of patients with metastatic CRC (mCRC). However, only patients with a wild-type KRAS may hope respond to EGFR inhibition. The majority of patients expresses oncogenic components of Ras signaling and thus have hyperactive Ras cascade operating independently from EGFR. Therefore, there is an urgent need to discover new Ras-independent oncogenic targets that regulate tumor cell invasion and metastasis in patients with mCRC. Using a pharmacological screen, we uncovered an additional TK that regulates CRC cell invasion and metastasis induction irrespective of KRAS mutation. Specifically, the clinically available TK inhibitor nilotinib (Tasigna) originally developed to target imatinib-resistant Bcr-Abl alleles in leukemia, also inhibits invasive properties of CRC cells. Nilotinib also strongly reduces liver metastases that were induced after injection of mCRC cells in the spleen of nude mice. Remarkably, the potency of nilotinib to inhibit invasive activity of mCRC cells was similar to the one observed in leukemia. Since we could not detect Abl deregulation in CRC cells, we speculated the involvement of an alternative target to be identified. Interestingly, a previous chemical-proteomic approach identified DDR1 as the highest affinity target of nilotinib (Rix et al, Blood, 110:4055-63). DDR1 is a poorly-characterized TK and a receptor for collagen, the major component of the extracellular matrix. We found that DDR1 is overexpressed in CRC and regulates CRC cell-induced invasion in vitro and liver metastasis formation in vivo. Finally, we have obtained evidence supporting that the pharmacological activity of nilotinib in CRC cells is mediated by inhibition of DDR1. Indeed, introduction of the nilotinib resistant DDR1 T701I mutant into CRC cells renders these cells resistant to nilotinib. Overall, our data suggest that targeting DDR1 by nilotinib may be a therapeutic value in patients with mCRC.

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 855. doi:1538-7445.AM2012-855
Abstract 862: Pre-clinical and molecular analysis of Crizotinib in c-Met positive uveal melanoma
Authors: de Lange Mark J. , Versluis Mieke , Luyten Gre P.M. , Jager Martine J. , van der Velden Pieter A.
Published: August 2012

Uveal melanoma (UM) is an intraocular neoplasm with an annual incidence of 7 per million. UM originates from melanocytes just like cutaneous melanoma (CM) and similar to CM, the RAS/RAF/MEK/ERK pathway is involved in the development of UM. However, not all UM seem to depend on the activation of this pathway. Loss of ERK signalling may be correlated with progression as the ERK negative cells are derived from UM metastasis. In these tumours, cells apparently use a different pathway to proliferate and survive. In order to identify treatment targets for metastasis patients we set out to identify the pathway which stimulates proliferation in the ERK negative UM cells. Growth factor receptor screening revealed that activation of c-Met or HGF receptor, is inversely correlated with ERK activation leaving the metastatic UM, halfway progression to metastasis, with both c-Met and ERK activated. In this study, we tested the efficacy of c-Met inhibitor Crizotinib and investigated primary and downstream targets to delineate the role of c-Met signalling in UM progression. Crizotinib was shown to specifically inhibit c-Met positive UM and this was most clear when cells were grown anchorage independent, in line with a role in dissemination for c-Met. With phospho-proteomics we showed that c-Met is the primary target of Crizotinib in UM while FAK emerges as the prime downstream target. Moreover, Src was shown to be the downstream kinase that transmits the c-Met signal to both FAK and ERK in metastatic UM. In UM metastasis, lacking active ERK, Src is absent and FAK is directly activated by c-Met. Combined, our data support the use of Crizotinib in late stage UM and reveals altered c-Met signalling in uveal melanoma progression.

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 862. doi:1538-7445.AM2012-862
Abstract 863: Tyrosine kinase activity profiling of metastatic malignant melanoma: Identification of possible therapeutic targets and markers predicting response to therapy
Authors: Tahiri Andliena , Roe Kathrine , Busch Christian , Lonning Per Eystein , Ree Anne H. , Kristensen Vessela N. , Geisler Juergen
Published: August 2012

Metastatic melanoma is an aggressive form of skin cancer that is highly resistant to conventional chemotherapy. Thus, novel approaches to improve the current treatment of advanced melanoma are urgently needed. Kinases are key regulators of cellular processes, and since nearly half of the tyrosine kinase complement is implicated in cancer, the information of kinase activity in tumors give important tumor-specific information on affected pathways and potential targets for treatment. With this in mind, we performed tyrosine kinase activity profiling of 26 tissue samples obtained from patients suffering from metastatic stage IV melanoma, prior to dacarbazine (DTIC) treatment, using Tyrosine Kinase PamChip® microarrays. Each sample lysate generated an individual phospho-substrate signature, representing the state of the kinase signalling cascades, which allowed us to compare affected targets in responders and non-responders to DTIC treatment. Further, we attempted to evaluate the kinase activity in response to BRAF inhibitor, PLX4032 (Vemurafenib), which has shown dramatic and positive effects in advanced melanoma tumors, and has recently been approved for the treatment of late stage melanoma. In this case, incubation was carried out on all samples with and without PLX4032. Our preliminary results showed that although the compound is mainly directed against a serine/threonine kinase, BRAF, PLX4032 also showed inhibitory effects on the activity of multiple tyrosine kinases. Since the drug is only beneficial for melanoma patients with BRAF V600E mutations, we compared the activated kinase pathways in patients with and without this mutation in order to identify alternative survival pathways in patients with wild type BRAF. The technology used in this study is based on a unique 3D flow-through microarray format that allows monitoring of the reaction in real time, ideal for the characterization of pathways. Each array contained 144 peptide substrates with sites for phosphorylation, representing about 100 different proteins. The use of kinase inhibitors will cause inhibition of corresponding kinases of activated signalling pathways; hence, providing us leads to therapeutic targets and possible prediction markers for response to therapy. The detailed effects of both PLX4032 and sunitinib in the chosen test system are currently prepared for presentation.

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 863. doi:1538-7445.AM2012-863