bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5446: Continuous real-time measurement of live and dead cells in culture over multiple days
Authors: Riss Terry , Vidugiriene Jolanta , Duellman Sarah , Zhou Wenhui , Vidugiris Gediminas , Valley Mike , Osterman Jean , Arbit Ruslan , Bernad Laurent , Meisenheimer Poncho , Cali James J. 
Published: August 2015

Abstract
Recently developed assay technologies have made it possible to use a multi-well plate reader to estimate the number of live or dead cells in culture in real time over a period of days to monitor the effects of long term exposure to test substances. An advantage of the real time assay chemistries is that the cells remain viable and available for other measurements. The purpose of this study was to determine if the real time assay methods could be multiplexed with other assay endpoints and applied to three dimensional (3D) culture models. The measurement of the number of live cells in real time is accomplished by providing a pro-substrate that viable cells (but not dead cells) can convert into a substrate for a luciferase derived from a marine shrimp. The pro-substrate and luciferase can be added with a single reagent addition step and because they are not toxic, they can be incubated with cells in culture for up to three days. Live cell conversion of the pro-substrate into a substrate for luciferase results in a luminescent signal that is proportional to the number of live cells. The signal diminishes immediately upon cell death. Detection of dead cells in real time is accomplished by using a fluorogenic DNA binding dye that is non-permeant and thus non-toxic to live cells. The dye enters dead cells that have a compromised membrane, binds to DNA, and becomes fluorescent. We have demonstrated that the real time assays can be multiplexed to measure both live and dead cells in the same sample. We further demonstrate the use of these assays to measure viability of cells in 3D spheroids formed using the hanging drop culture technique. We also demonstrate multiplexing of real time assays with subsequent measurement of apoptosis or extraction of RNA for expression studies. Utilization of real time assay chemistries that are non-toxic enabled repeated recording of data in a kinetic mode from the same sample of cells which eliminated the need for duplicate cultures and provided flexibility during assay protocol development. The ability to multiplex real time assays improved overall lab efficiency by eliminating the need for duplicate cultures for each experiment.
Citation Format: Terry Riss, Jolanta Vidugiriene, Sarah Duellman, Wenhui Zhou, Gediminas Vidugiris, Mike Valley, Jean Osterman, Ruslan Arbit, Laurent Bernad, Poncho Meisenheimer, James J. Cali. Continuous real-time measurement of live and dead cells in culture over multiple days. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5446. doi:10.1158/1538-7445.AM2015-5446
Abstract 5463: Role of the endothelin axis in astrocyte and endothelial cell mediated chemoprotection of cancer cells
Authors: Kim Mark S. , Choi Hyun Jin , Lee Ho-Jeong , He Junqin , We Qiuyu , Langley Robert R. , Kim Sin-Jin , Fidler Isaiah J. 
Published: August 2015

Abstract
Background. Recent evidence suggests that astrocytes protect cancer cells from chemotherapy by stimulating the upregulation of anti-apoptotic genes in cancer cells. We investigated the possibility that activation of the endothelin axis orchestrates survival gene expression and chemoprotection in MDA-MB-231 breast cancer cells and H226 lung cancer cells.
Methods. Cancer cells, murine astrocytes, and murine fibroblasts were cultured in isolation and expression of endothelin (ET) peptides and ET receptors (ETAR and ETBR) was compared with expression on cancer cells and astrocytes (or cancer cells and fibroblasts) that were co-incubated for 48 hours. Type-specific endothelin receptor antagonists were used to evaluate the contribution of ETAR and ETBR to astrocyte-induced activation of the protein kinase B (AKT)/mitogen-activated protein kinase (MAPK) signal transduction pathways, anti-apoptotic gene expression, and chemoprotection of cancer cells. We also investigated the chemoprotective potential of brain endothelial cells and microglial cells.
Results. Gap junction signaling between astrocytes and MDA-MB-231 cancer cells stimulates upregulation of interleukin 6 (IL-6) and IL-8 expression in cancer cells, which increases ET-1 production from astrocytes and ET receptor expression on cancer cells. ET-1 signals for activation of AKT/MAPK and upregulation of survival proteins that protect cancer cells from taxol. Brain endothelial cell-mediated chemoprotection of cancer cells also involves endothelin signaling. Dual antagonism of ETAR and ETBR is required to abolish astrocyte- and endothelial cell-mediated chemoprotection.
Conclusions. Bidirectional signaling between astrocytes and cancer cells involves upregulation and activation of the endothelin axis, which protects cancer cells from cytotoxicity induced by chemotherapeutic drugs.
Citation Format: Mark S. Kim, Hyun Jin Choi, Ho-Jeong Lee, Junqin He, Qiuyu We, Robert R. Langley, Sin-Jin Kim, Isaiah J. Fidler. Role of the endothelin axis in astrocyte and endothelial cell mediated chemoprotection of cancer cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5463. doi:10.1158/1538-7445.AM2015-5463
Abstract 5475: Cancer exosome promotes cisplatin resistance In bladder cancer and inhibition of exosome sensitizes bladder cancer cells to cisplatin chemotherapy
Authors: Wu Chiahao , Silvers Chris , Guancial Elizabeths , Hsu Jong-Wei , Messing Edward , Lee Yi-Fen 
Published: August 2015

Abstract
Exsosomes are nano-vesicles secreted by most types of cells, and substantially increased secretion in cancer cells. Their role in tumorigenesis/cancer progression have been extensively studied but their role in regulating therapy response have been suggested without in depth investigation. We are particularly interested in exosome's roles in cisplatin resistant, since cisplatin is one of the most widely used chemodrugs for many cancer types.
To investigate the role of exosomes in cisplatin resistance in bladder cancer, we examined the cisplatin sensitivity among five bladder cancer cell lines. The expression levels of exosome associated genes that are involved in exosome biogenesis and secretion/internalization were examined by quantative PCR. Interestingly, cisplatin resistant bladder cancer cells express relative high level of exosome associated genes as compared to cisplatin sensitive cells. More importantly, cisplatin treatment induces the expression of exosome associated genes’ expression in cisplatin resistant cells, but not in sensitive cell. Among all those genes, we are particularly interested in Rab gene family, and mechanistic study of targeting those Rab proteins is undergoing. The correlative exosome genes profiles with cisplatin resistance suggested exosome production might facilitate cancer cells escaping from cisplatin-induced cell death; thereby, inhibition of exosome might sensitize cancer cells response to cisplatin. To test this, we have screened several compounds that are known to inhibit exosome biogenesis and uptake, and. found that inhibition of exosomes by dynasore + NSC23766, indeed can sensitize bladder cancer cells to cisplatin. In summary, we concluded that cancer exosome play a critical role of promoting cisplatin resistance in bladder cancer, and inhibition of exosomes sensitizes bladder cancer cell response to cisplain. The exosome related profiles potentially can serve as therapy biomarkers
Citation Format: Chiahao Wu, Chris Silvers, Elizabeths Guancial, Jong-Wei Hsu, Edward Messing, Yi-Fen Lee. Cancer exosome promotes cisplatin resistance In bladder cancer and inhibition of exosome sensitizes bladder cancer cells to cisplatin chemotherapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5475. doi:10.1158/1538-7445.AM2015-5475
Abstract 548: Exosome as biomarkers and diagnostics in bladder cancer
Authors: Liu Yu-Ru , Silver Christopher , Lee Yi-Fen 
Published: August 2015

Abstract
Esosomes are small membrane vesicles secreted by most cell types. They are first found in reticulocytes and play important roles in cell-to-cell communication. Recent studies suggest that exosomes secreted by tumor cells containing tumor specific proteins and miRNAs.
Previous study shows that exosomes purified from high grade bladder cancer promote cancer progression. This study aims to characterize the genetic contents in bladder cancer exosomes through mass spectrometry and miRNA array. Exosomes were purified from bladder cancers, immortalized bladder cells, and urine of healthy volunteers and muscle invasive bladder cancer patients. Via proteomics analysis, we have identified several cancer associated exosomal proteins that were found only in bladder cancer, including EDIL-3, an anagiogenic protein in our previous report. Functional characterization of those proteins is currently undertaken with a focus on periostin an extracellulear matrix protein.
In addition to protein, exosomes also contain miRNAs that regulate cellular behaviors. Thus, we performed miRNA expression profiling microarray on exosomes from high grade bladder cancer cells, and immortalized bladder cells (LC Sciences). The differences in miRNA expression were compared and differences were determined by a student T test (P&lt;0.05). Results showed significant difference miRNAs between cancer cells vs normal cells. Some of cancer-specific miRNAs which have shown to play critical roles in tumorigenesis and cancer progression will be study priority. Ultimately our work should contribute to the understanding of how exosomes contribute to bladder cancer biology and may elucidate novel urine exosomes markers for bladder cancer diagnosis.
Note: This abstract was not presented at the meeting.
Citation Format: Yu-Ru Liu, Christopher Silver, Yi-Fen Lee. Exosome as biomarkers and diagnostics in bladder cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 548. doi:10.1158/1538-7445.AM2015-548
Abstract 5482: CYP2D6 genotype and response to neoadjuvant tamoxifen therapy: a prospective study in Japan
Authors: Zembutsu Hitoshi , Nakamura Seigo , Akashi Sadako , Kuwayama Takashi , Watanabe Chie , Takei Hiroyuki , Ishikawa Takashi , Hasegawa Yoshie , Lee Soo Chin , Yu Tan Ern , Matsukata Ayami , Matsumoto Hiroshi , Kutomi Goro , Nakamura Yusuke 
Published: August 2015

Abstract
Although lots of CYP2D6-Tamoxifen studies have been reported, the results of the association are still controversial. Because prospective studies are necessary to fully establish the value of CYP2D6 genotyping in tamoxifen therapy, we have initiated the prospective study to clarify the relationship between CYP2D6 genotype and response to preoperative Tamoxifen therapy using a surrogate marker of Ki-67 labeling index in cancer tissue (C-GENT study) since July, 2012. Of the 342 patients who have been recruited in this study, we analyzed 173 cases whose genotyping and measurement of Ki-67 Labeling Index were completed. We observed significant Ki-67 decrease after 2-4 weeks preoperative Tamoxifen therapy (P = 0.000052). Moreover, the patients with CYP2D6 *V/*V is unlikely to show Ki-67 decrease compared with those with CYP2D6 Wt/Wt or Wt/*V (P = 0.00046). Although further analysis should be continuing using a larger number of samples registered in this study, our results suggested that polymorphisms in CYP2D6 could be a key predictor for the prognosis of patients with breast cancer treated with tamoxifen.
Citation Format: Hitoshi Zembutsu, Seigo Nakamura, Sadako Akashi, Takashi Kuwayama, Chie Watanabe, Hiroyuki Takei, Takashi Ishikawa, Yoshie Hasegawa, Soo Chin Lee, Tan Ern Yu, Ayami Matsukata, Hiroshi Matsumoto, Goro Kutomi, Yusuke Nakamura. CYP2D6 genotype and response to neoadjuvant tamoxifen therapy: a prospective study in Japan. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5482. doi:10.1158/1538-7445.AM2015-5482
Abstract 5484: FGFR2 amplification is predictive of sensitivity to regorafenib in gastric and colorectal cancers
Authors: Cha Yongjun , Kim Hwang-Phill , Han Sae-won , Yun Jiyeon , Song Sang-Hyun , Kim Tae-You 
Published: August 2015

Abstract
Purpose Regorafenib is an orally bioavailable, small molecular multikinase inhibitor of VEGFR, PDGFR, FGFR, KIT, RET, BRAF, BRAFV600E, CRAF, and TIE2. Although regorafenib has demonstrated survival benefits in patients with metastatic colorectal and gastrointestinal stromal tumors, there is no proven biomarker predicting sensitivity to regorafenib. We investigated preclinical activity of regorafenib in gastric and colorectal cancer cell-lines to identify genetic alterations associated with sensitivity to regorafenib.
Methods Fourteen gastric cancer cell lines and 18 colorectal cancer cell lines were screened for regorafenib sensitivity with cell proliferation assays. We performed targeted exome and transcriptome sequencing of the cell lines to identify genetic alterations associated with sensitivity to regorafenib. Mechanisms of regorafenib sensitivity were investigated in regorafenib sensitive cell lines with immunoblotting and flow cytometry.
Results Two gastric (SNU-16, KATO-III) and 1 colorectal (NCI-H716) cancer cells were sensitive to regorafenib with GI50 values of less than 1 μM. Mutation profiles and mRNA expression levels of regorafenib target molecules in regorafenib sensitive and insensitive cell lines demonstrated that amplification of FGFR2 was the only genetic alteration associated with in vitro sensitivity to regorafenib. FGFR2 amplified gastric (SNU-16, KATO-III) and colorectal (NCI-H716) cancer cell lines revealed activated signaling through FGFR2 pathway, and knock-down of FGFR2 with siRNA significantly repressed cellular growth rate and decreased sensitivity to regorafenib. Regorafenib effectively inhibited phosphorylation of FGFR2 and its downstream signaling molecules in dose-dependent manner selectively in FGFR2 amplified cells. In cell cycle analysis, regorafenib induced G1/S arrest (SNU-16, KATO-III) and apoptosis (NCI-H716), however no significant changes were seen in cell lines without FGFR2 amplification.
Conclusions Signaling through FGFR2 pathway plays a key role in cellular growth and survival in FGFR2 amplified cancers. Regorafenib effectively abrogates activated FGFR2 signaling in FGFR2 amplified gastric and colorectal cancer cells, and therefore should be considered for integration into treatment in patients with FGFR2 amplified gastric and colorectal cancers.
Citation Format: Yongjun Cha, Hwang-Phill Kim, Sae-won Han, Jiyeon Yun, Sang-Hyun Song, Tae-You Kim. FGFR2 amplification is predictive of sensitivity to regorafenib in gastric and colorectal cancers. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5484. doi:10.1158/1538-7445.AM2015-5484
Abstract 5501: Antagonism of sorafenib and regorafenib actions by epidermal growth factor in hepatoma cells
Authors: Carr Brian I. , D'Alessandro Rosalba , Refolo Maria Grazia , Lippolis Catia , Messa Caterina , Cavallini Aldo 
Published: August 2015

Abstract
Background. Platelet numbers are related to growth and aggressiveness of several tumor types, including hepatoma (hepatocellular carcinoma). We previously found that platelet lysates also stimulated growth and migration of hepatoma lines in vitro and antagonized the growth-inhibitory effects on hepatoma cells of both multikinase inhibitors sorafenib and regorafenib (BMC Cancer 2014; 14:351).
Aims. To investigate the mechanisms for the platelet-mediated multikinase drug resistance, we examined the actions of several mitogens that are known to be contained by platelets (PDGF, VEGF, TGF beta, FGF, IGF-1, EGF).
Results. We found that epidermal growth factor (EGF) alone antagonized the growth inhibitory actions of Sorafenib or Regorafenib on HepG2, PLC/PRF/5 and Hep3B human hepatoma cells, using an MTT assay. Detailed studies revealed that EGF also antagonized Regorafenib-mediated inhibition of cell migration and invasion. Furthermore, EGF also blocked regorafenib-induced apoptosis, measured by Annexin V using a MUSE cell analyzer. These effects of EGF were antagonized by concomitant addition to the cultures of EGF receptor antagonist Erlotinib, showing that the EGF receptor was involved in the mechanisms of EGF-mediated blocking of Regorafenib effects. In addition, Erlotinib also partially blocked the effects of platelet lysates in antagonizing regorafenib-mediated growth inhibition. Similar antagonism was found using IGF-1, and those effects were blocked by GSK IGF-1 receptor antagonist. Other platelet factors had only weak actions in these assays.
Conclusions. EGF antagonized regorafenib-mediated growth and migration inhibition and apoptosis induction in hepatoma cells. These EGF actions were mediated via the EGF receptor, as were the similar platelet lysate actions.
Citation Format: Brian I. Carr, Rosalba D'Alessandro, Maria Grazia Refolo, Catia Lippolis, Caterina Messa, Aldo Cavallini. Antagonism of sorafenib and regorafenib actions by epidermal growth factor in hepatoma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5501. doi:10.1158/1538-7445.AM2015-5501
Abstract 5507: Characterization of ADC bystander killing in admixed tumor model
Authors: Li Fu , Emmerton Kim Kissler , Jonas Mechthild , Zhang Xinqun , Law Che-Leung 
Published: August 2015

Abstract
We have previously described an admixed tumor model, consisting of the CD30+ ALCL cell line Karpas 299 and CD30- Karpas-35R (SGN-35 resistant) cells, to study the phenomenon of ‘bystander killing’ of antigen negative tumor cells by antibody-drug conjugates (ADCs) in tumors with a heterogeneous target expression profile. Here we study the contribution of payload membrane permeability and target expression in bystander killing.
We hypothesize that membrane permeability of payloads may impact bystander killing in addition to their intrinsic potency. Monomethyl auristatin E (MMAE)-based ADCs utilizing the cleavable linkers valine-citrulline (vc) or glucuronide (gluc) release the membrane permeable MMAE. As a result, antiCD30-vcMMAE and antiCD30-glucMMAE showed similar bystander killing in vivo. We then compared the more membrane permeable payloads, MMAE and pyrrolobenzodiazepine dimer (PBD), to those that are less permeable, monomethyl auristatin F (MMAF) and auristatin T (AT), as antiCD30 conjugates in this model. We found that antiCD30-AT and antiCD30-vcMMAF killed only CD30+ tumor cells in admixed tumors, while antiCD30-MMAE, antiCD30-GlucMMAE and antiCD30-PBD killed both CD30+ and CD30- cells. These results suggest that membrane permeability is one of the key factors determining in vivo bystander killing.
To elucidate the role of antigen positive cells in bystander killing in this model, we varied the implant ratio of CD30+ cells and evaluated ADC activity in vivo. Flow cytometric analysis and CD30 immunohistochemistry confirmed a corresponding change of CD30+ cell percentage in the resulting admixed tumors (∼40%, 70%, or 80%). Treatment with ADCs in these tumors demonstrated that antiCD30-vcMMAE induced remissions in tumors with 70 or 80% CD30+ cells, whereas antiCD30-PBD produced remissions in all three admixed tumor ratios.
Collectively, these data confirm that membrane permeable payloads can kill antigen negative cells in vivo. We hypothesize that the enhanced bystander killing by PBD-based ADCs may result from the intrinsic potency of the small molecule and/or membrane permeability. This information will be useful to consider when designing future ADCs.
Citation Format: Fu Li, Kim Kissler Emmerton, Mechthild Jonas, Xinqun Zhang, Che-Leung Law. Characterization of ADC bystander killing in admixed tumor model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5507. doi:10.1158/1538-7445.AM2015-5507
Abstract 551: Identification of gene-expression biomarkers in urine pathology specimens for the detection of bladder cancer
Authors: Chapman Karen B. , Qiu Liqun , Kidd Jennifer , Baxi Aparna , Lachter Markus D. , Wagner Joseph , Rosenthal Dorothy L. , Olson Matthew T. 
Published: August 2015

Abstract
Bladder cancer recurrence screening is typically managed with a combination of urine cytology and cystoscopy, each with its inherent caveats. Cystoscopy is generally considered the gold standard for diagnosing bladder cancer and can yield a valuable biopsy, but is too invasive and expensive for routine screening and surveillance. Voided urine cytology is inexpensive and readily available, but lacks the desired level of sensitivity. Furthermore, up to 20% of urine cytology specimens fall into one of two indeterminate categories that require follow-up testing, typically in the form of an invasive cystoscopy procedure. With the goal of developing a urine-based molecular test for bladder cancer recurrence surveillance, we examined gene expression biomarkers in urine cytopathology specimens. 90 patient urine samples were analyzed: 45 high-grade urothelial carcinoma (HG-UC) and 45 benign samples. All patient samples were submitted to the Johns Hopkins Cytopathology Department for evaluation for bladder cancer via urine cytology. The samples represent patients undergoing bladder cancer recurrence screening and patients without a prior history of bladder cancer presenting with hematuria. The urine sediments were stored in liquid based cytology solution at 4°C for a period of 7 - 10 days, during which time the cytopathology diagnosis was determined from a portion of the sample. Post-diagnosis, the pathology sample “leftover” was analyzed for gene expression. RNA was extracted, amplified and microarray analysis was performed and a panel of molecular biomarkers was identified. Individual genes in this panel discriminate between HG-UC and benign in this training set with an average ROC of 0.86 (p &lt;0.0001) while a combined panel of 10 genes yields a ROC of 0.97 (p &lt;0.0001). Differential expression of some of the individual genes from the panel was confirmed by qPCR. Additional evaluation of this gene panel in a validation set is ongoing and results from this validation study will be discussed.
Citation Format: Karen B. Chapman, Liqun Qiu, Jennifer Kidd, Aparna Baxi, Markus D. Lachter, Joseph Wagner, Dorothy L. Rosenthal, Matthew T. Olson. Identification of gene-expression biomarkers in urine pathology specimens for the detection of bladder cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 551. doi:10.1158/1538-7445.AM2015-551