bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2539: Survival of lymphoma and beverages consumptions; alcohol, tea and coffee: results from National Cancer Centre, Singapore lymphoma cohort.
Authors: Lee Yin Leng , Tay Kevin Kuang Wei , Tan Soo Yong , Tan Leonard , Quek Richard Hong Hui , Tao Miriam , Ha Tam Cam , Lim Soon Thye
Published: August 2014

Background: In 2008, Singapore Cancer Registry reported lymphoma age-standardized rate (ASR) among males and females was 11.2 per 100,000 and 7.5 per 100,000, respectively with highest incidence observed in Malays, followed Chinese and Indians. Lymphoma incidence rates have been increasing since 1968 (female; ASR: 1.8 per 100,000) and is now the seventh and ninth most common cause of cancer for both men and women, respectively. Lymphoma is widely studied but the cause is yet to be ascertained. Epidemiological studies have shown modifiable behaviour such as heavy alcohol use results in poorer survival across various cancer types in Western population while reviews of tea and coffee consumptions in association with survival are limited. Here, we examined the impact of beverages consumption; alcohol, tea and coffee on lymphoma survival in South East Asia.

Methods: A cohort of 384 lymphoma patients with confirmed histology was recruited from January 2004 to December 2009 and was prospectively followed through till July 2012. At diagnosis, lifestyle characteristics were assessed using a structured questionnaire. Cox proportional hazards regression was used to estimate the prognostic value of three beverages; alcohol, tea and coffee, on lymphoma survival adjusted for other clinical prognostic factors; LDH, ECOG, Ann Arbor stage and extra-nodal sites.

Results: The overall median follow-up was 4.2 years (range: 0.02 to 8.3). Among all lymphoma subtypes, the most prevalent was DLBCL, 183 (47.7%) with highest incidence among Chinese, 305 (79.4%) followed by Malay 53 (13.8%) and Indian 24 (6.3%). Alcohol consumptions were associated with poorer survival for all lymphoma (ptrend=0.004) with increasing alcohol use whereas female black tea (plog-rank=0.0063) and coffee (plog-rank=0.018) consumptions had lower risk of death compared to non-drinkers. Full model including alcohol (one or more drinks per day; HR=2.35, 95% CI: 1.38-4.00, p=0.002) and clinical prognostic factors results in c index, 0.75 and 0.72, for overall survival and progression free survival, respectively.

Conclusion: Alcohol consumption in our cohort is predictive of survival while tea and coffee had a weak survival association among lymphoma. This model warrant external validation.

Citation Format: Yin Leng Lee, Kevin Kuang Wei Tay, Soo Yong Tan, Leonard Tan, Richard Hong Hui Quek, Miriam Tao, Tam Cam Ha, Soon Thye Lim. Survival of lymphoma and beverages consumptions; alcohol, tea and coffee: results from National Cancer Centre, Singapore lymphoma cohort. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2539. doi:10.1158/1538-7445.AM2013-2539
Abstract 2541: A prospective study of medical diagnostic x-rays and risk of thyroid cancer.
Authors: Neta Gila I. , Rajaraman Preetha , Berrington de Gonzalez Amy , Doody Michele , Alexander Bruce , Simon Steve , Freedman D. Michal , Linet Martha , Sigurdson Alice
Published: August 2014

While diagnostic x-rays provide important medical benefits, cancer risks associated with their exposure are also possible. The thyroid gland is the most radiosensitive organ, and radiotherapy to the head and neck is associated with an increased risk of thyroid cancer. However, it is not known whether diagnostic x-rays, which yield a lower dose of ionizing radiation to the thyroid gland than radiotherapy, also increase the risk. The United States Radiological Technologists study (1983-2006) is a nationwide, prospective cohort with extensive questionnaire data on history of personal diagnostic imaging procedures collected prior to cancer diagnosis. Cox proportional hazards were used to estimate thyroid cancer (TC) risks related to the number and type of selected procedures. Potential modifying effects of age and calendar year of the first x-ray in each category of procedures were assessed. Incident thyroid cancers (n=251) were diagnosed among 75,494 technologists (1.3 million person-years, mean follow-up=17 years). Overall, there was no clear evidence of TC risks associated with diagnostic x-rays, except for dental x-rays. The authors observed a 13% increase in TC risk for every 10 reported dental x-rays (95%CI: 1.01-1.26), driven by dental x-rays first received before 1970, but no evidence that the relationship between dental x-rays and TC was associated with childhood or adolescent exposures as would have been anticipated. The lack of association of TC with x-rays which expose the thyroid to higher radiation doses than dental x-rays underscores the need to conduct a detailed radiation exposure assessment to enable quantitative evaluation of risk.

Citation Format: Gila I. Neta, Preetha Rajaraman, Amy Berrington de Gonzalez, Michele Doody, Bruce Alexander, Steve Simon, D. Michal Freedman, Martha Linet, Alice Sigurdson. A prospective study of medical diagnostic x-rays and risk of thyroid cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2541. doi:10.1158/1538-7445.AM2013-2541

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2581: Intervention of chronically-induced human breast cell carcinogenesis.
Authors: Choudhary Shambhunath , Sood Shilpa , Wang Hwa-Chain Robert
Published: August 2014

The purpose of our study was to investigate the ability of dipyridamole (DPM) [2,6-bis(diethanolamino)-4,8-dipiperidinopyrimido-(5,4-d)-pyrimidine] to intervene in breast cell carcinogenesis induced by chronic exposure to environmental carcinogens. More than 85% of breast cancers are sporadic and attributable to long-term exposure to environmental carcinogens through a multistep and multiyear disease process. Therefore, it is urgent to identify preventive agents effective in intervention of cellular carcinogenesis. DPM, an antioxidant and a synthetic inhibitor of phosphodiesterase, is approved by the Food and Drug Administration to inhibit platelet aggregation and vasoconstriction. DPM is also able to enhance the cytotoxicity of anticancer agents, such as cisplatin, adriamycin, and 5-fluorouracil, in various cell types. However, its ability to block breast cell carcinogenesis has not been determined. Using our chronically-induced breast cell carcinogenesis model as a target system, we found that DPM, at a non-cytotoxic physiologically-achievable dose of 10 nmol/L, effectively blocked breast cell carcinogenesis induced by chronic exposure to three unrelated carcinogens: 2-amino-1-methyl-6-phenylimidazo[4,5- b]pyridine (PhIP), benzo[a]pyrene (B[a]P), and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). The ability of DPM to block H-Ras upregulation, thus blocking ERK pathway activation, reactive oxygen species (ROS) elevation, and DNA damage in each exposure, may account for its mechanisms in intervention of carcinogenesis induced by chronic exposures to PhIP. Likewise, DPM's ability to block ROS elevation and DNA damage may account for its mechanisms in intervention of carcinogenesis chronically induced by NNK and B[a]P, as well. Our study revealed, for the first time, the novel ability of DPM to block breast cell carcinogenesis induced by three unrelated carcinogens. DPM should be seriously considered as a chemopreventive agent in development of strategies for reducing the risk of sporadic breast cancer associated with long-term exposure to environmental carcinogens.

Citation Format: Shambhunath Choudhary, Shilpa Sood, Hwa-Chain Robert Wang. Intervention of chronically-induced human breast cell carcinogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2581. doi:10.1158/1538-7445.AM2013-2581
Abstract 2599: Inhibitory effects of new 1H-pyrrolo[3,2-c]pyridine derivative, KIST101029, on AP-1 activity and neoplastic cell transformation induced by insulin-like growth factor 1.
Authors: Yun Hyo Jeong , Choi Hong Seok
Published: August 2014

Diarylureas and diarylamides derivatives are reported to have antitumor activity. Encouraged by the interesting antiproliferative activity of diarylurea and diarylamide derivatives, we synthesized a new series of diarylureas and diarylamides containing pyrrolo[3,2-c]pyridine scaffold. In this study, we demonstrate that a new 1H-pyrrolo[3,2-c]pyridine derivative, KIST101029, inhibits neoplastic cell transformation induced by insulin-like growth factor 1 (IGF-1) in mouse epidermal JB6 Cl41 cells. The KIST101029 compound inhibited mitogen-activated protein kinase/extracellular signal-regulated kinase kinases (MEK), c-jun N-terminal kinases (JNK), and mechanistic target of rapamycin (mTOR) signaling pathways induced by IGF-1 in JB6 Cl41 cells, resulting in the inhibition of c-fos and c-jun transcriptional activity. In addition, the KIST101029 inhibited the associated activator protein-1 (AP-1) transactivation activity and cell transformation induced by IGF-1 in JB6 Cl41 cells. Consistent with these observations, in vivo chorioallantoic membrane assay also showed that the KIST101029 inhibited IGF-1-induced tumorigenicity of JB6 Cl41 cells. Taken together, these results indicate that a new 1H-pyrrolo[3,2-c]pyridine derivative, KIST101029, might exert chemopreventive effects through the inhibition of phosphorylation of MAPK and mTOR signaling pathway.

Citation Format: Hyo Jeong Yun, Hong Seok Choi. Inhibitory effects of new 1H-pyrrolo[3,2-c]pyridine derivative, KIST101029, on AP-1 activity and neoplastic cell transformation induced by insulin-like growth factor 1. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2599. doi:10.1158/1538-7445.AM2013-2599

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2600: Polyunsaturated omega-3 fatty acids and their metabolites preferentially inhibit cell proliferation and motility in triple negative over luminal breast cancer cells.
Authors: Pogash Thomas J. , Lopez de Cicco Ricardo , Pressly Benjamin , Russo Irma H. , Himmelberger Julie A. , Amin Shantu , Gowda Krishne , El-Bayoumy Karam , Manni Andrea , Russo Jose
Published: August 2014

The role of diet as a preventive measure in breast cancer remains controversial. Current literature suggests that the risk of developing breast cancer may decrease or increase with the intake of omega-3 (n-3) and omega-6 (n-6) fatty acids, respectively. Omega 3 polyunsaturated fatty acids (PUFAs) exert an anticancer effect by affecting multiple cellular mechanisms leading to inhibition of proliferation and induction of apoptosis. It is well known that breast cancer comprises distinct molecular subtypes which differ in their responsiveness to therapeutic and preventive agents. In these experiments, we tested the hypothesis that n-3FA may preferentially affect triple negative breast cancer cells for which no targeted intervention is presently available. The in vitro effects of n-3 PUFA Docosahexaenoic acid (DHA) and its metabolites, 4-OH-DHA and 4-OXO-DHA, were tested on proliferation and motility of seven triple negative human basal breast cell lines at different stages of transformation (MCF-10F, trMCF, bsMCF, caMCF, MDA-MB-231, Hs-578T, and BT-549), and three luminal breast cancer cell lines (MCF-7, T-47D, and SK-BR-3). Cell proliferation was measured with the tetrazolium MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) assay and cell migration was assessed using the wound healing assay via time lapse microscopy with MetaMorph 7.7.7.8. All assays were performed in quadruplicate. Statistical significance was assessed using Student's t-test (p<0.05). DHA and its metabolites significantly inhibited cell proliferation (20-90% reduction) of both basal and luminal breast cancer cell lines. The inhibitory effect was more pronounced on triple negative basal breast cancer cell lines as compared to luminal breast cancer cell lines. DHA did not produce an inhibitory effect on cell migration. However, the metabolites of DHA significantly reduced (20-60% reduction) cell migration on the bsMCF cell line. Our data provide novel information regarding the preferential antitumor effect of DHA and its metabolites on basal type breast cancer. (This project was supported by the Komen Foundation (KG081632).

Citation Format: Thomas J. Pogash, Ricardo Lopez de Cicco, Benjamin Pressly, Irma H. Russo, Julie A. Himmelberger, Shantu Amin, Krishne Gowda, Karam El-Bayoumy, Andrea Manni, Jose Russo. Polyunsaturated omega-3 fatty acids and their metabolites preferentially inhibit cell proliferation and motility in triple negative over luminal breast cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2600. doi:10.1158/1538-7445.AM2013-2600
Abstract 2624: Sodium butyrate induced cellular senescence and inhibited invasion of cancer cells with distinct mechanism.
Authors: Itoh Kazuyuki , Yoshioka Kiyoko , Nakagawa Hidemitsu
Published: August 2014

Sodium butyrate (SB) -C-4 saturated fatty acid (short-chain fatty acids) present in the human bowel membrane in the quite high concentration (2 mM) as food metabolites. It was previously reported that SB showed anti-tumor effect as HDAC inhibitor, however, the precise mechanism has not been elucidated yet. Here, we focused on the role of SB on cancer cell growth, motility and invasion. Physiological concentration of SB (0.25-4 mM) induced inhibition of cell growth (2D), colony formation in soft agar, in vitro motility and invasiveness with Boyden chamber and wound healing assay in a dose-dependent fashion using human fibrosarcoma (HT1080), glioblastoma (A172) cells and rat mammary Walker 256 cells. SB also affected the morphology of HT1080, A172 and Walker 256 cells, namely spread out with marked stress fibers bridging focal adhesions. Phosphorylation levels of focal adhesion kinase (Y577 and Y397 sites) were increased but that of myosin light chain (S19 site) was not altered. All of these biological effects of SB were reversible, and recovered after withdrawal. In contrast, HDAC inhibitor trichostatinA inhibited cancer cell growth irreversibly but not affected motility, invasiveness and morphology. In addition, HT1080 and A172 cells treated with SB showed positivity for senescence-associated β-gal (SA-β gal) staining with elevated expression levels of p53 and p21 proteins in a time and dose-dependent manner. In contrast, mRNA level of p21 was sharply declined after the treatment with SB. Knocking down of p21 protein expression by siRNA reversed both the growth inhibition and positivity for SA-β gal staining but not the inhibition of invasiveness. Thus p21 was required for cellular senescence but not for the decrease in motility induced by SB. Further, proteasome inhibitor, MG132 decreased the cellular growth and inhibited motility and invasiveness of the HT1080 and A172 cells, and showed similar all these biological effects of SB. Finally, intrathecal administration of SB effectively inhibited the invasion of Walker256 cells into the brain and elongated the animal (rat) survival in vivo. Collectively, SB inhibited growth with induced cellular senescence, inhibited invasion by distinct mechanism, and thus would be a good candidate for anti-invasive therapy without severe adverse effects.

Citation Format: Kazuyuki Itoh, Kiyoko Yoshioka, Hidemitsu Nakagawa. Sodium butyrate induced cellular senescence and inhibited invasion of cancer cells with distinct mechanism. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2624. doi:10.1158/1538-7445.AM2013-2624
Abstract 2629: PIK3CA mutation promotes tumor cell invasion and migration independently from AKT and mTOR kinase in cancer cells.
Authors: Bhattachara Bhaskar , Stasik Enrico , Low Sarah , Sturz Andrea , Schatz Christoph , Soong Richie , Liu Ningshu
Published: August 2014

Although the role of phosphoinositide-3 kinase (PI3K) in promoting tumorigenesis, tumor cell survival, proliferation and metabolism has been extensively evaluated, the mechanisms of other oncogenic processes mediated by PI3K have not been fully addressed. We report here a novel molecular mechanism by which PI3K mediates tumor cell invasion and migration using isogenic cell lines and highly selective PI3K pathway inhibitors. First of all, using a set of MCF10A-derived isogenic cell lines with PIK3CAH1047R, PTEN-deletion, or AKTE17K mutations, we found that activating the PIK3CA mutation, but not PTEN-deletion or AKT mutation, promoted tumor cell migration in a wound healing assay. Treatment of the PIK3CAH1047R isogenic cell line with a highly selective PI3K inhibitor (BAY 80-6946) effectively blocked tumor cell migration. Interestingly, neither allosteric AKT inhibitor nor mTOR kinase inhibitors were able to inhibit PIK3CA-stimulated cell migration although both AKT and mTOR kinase inhibitors potently inhibited AKT phosphorylation in these cells. In contrast, AKT inhibition enhanced the migration of PIK3CA mutant isogenic cells. This result revealed a novel AKT- and mTOR-independent mechanism by which activating PI3K mutation stimulates tumor cell migration. This conclusion was further confirmed in a panel of gastric cancer cell line in both wound healing and migration assays. Additional mechanistic experiments are being performed to examine downstream signaling events and will be presented. Taken together, these data suggest a role of PI3K in contributing to tumour metastasis independent of the canonical PI3K-AKT-mTOR axis and further validation of using AKT inhibitor for the treatment of metastatic PIK3CA mutant tumors to be conducted.

Citation Format: Bhaskar Bhattachara, Enrico Stasik, Sarah Low, Andrea Sturz, Christoph Schatz, Richie Soong, Ningshu Liu. PIK3CA mutation promotes tumor cell invasion and migration independently from AKT and mTOR kinase in cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2629. doi:10.1158/1538-7445.AM2013-2629
Abstract 2631: IL-6 promotes ICAM-1 expression and cell motility in human osteosarcoma.
Authors: Fong Yi-Chin , Tang Chih Hsin
Published: August 2014

Osteosarcoma is characterized by a high malignant and metastatic potential. Interleukin-6 (IL-6) is a multifunctional cytokine that is associated with the disease status and outcomes of cancers. However, the effect of IL-6 on migration activity in human osteosarcoma cells is mostly unknown. Here we found that IL-6 increased the migration and expression of intercellular adhesion molecule-1 (ICAM-1) in human osteosarcoma cells. Transfection of cells with ICAM-1 siRNA reduced IL-6-mediated cell migration. We also found that expression of IL-6 was significantly greater in human osteosarcoma tissues than in normal bone. The integrin-linked kinase (ILK)/Akt/AP-1 pathway was activated after IL-6 treatment, and IL-6- induced expression of ICAM-1 and migration activity was inhibited by the specific inhibitor and siRNA of ILK, Akt, and AP-1 cascades. In addition, over-expression of IL-6 shRNA inhibited the migratory ability and ICAM-1 expression in osteosarcoma cells. Taken together, these results indicate that IL-6 and IL-6 receptor interaction occurs through ILK and Akt, which in turn activates AP-1, resulting in the activations of ICAM-1 and contributing the migration of human osteosarcoma cells.

Citation Format: Yi-Chin Fong, Chih Hsin Tang. IL-6 promotes ICAM-1 expression and cell motility in human osteosarcoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2631. doi:10.1158/1538-7445.AM2013-2631
Abstract 2635: Galectin-1 modulates glioblastoma cell migration via interaction with FAK.
Authors: Velpula Kiran Kumar , Bhasin Arnima , Zhang Jane S. , Tsung Andrew J. , Veeravalli Krishna Kumar , Rao Jasti S. , Dasari Venkata Ramesh
Published: August 2014

Malignant glioblastomas (GBM) are the most common brain tumors and exhibit excessive growth, cell cycle regulation, neovascularization, angiogenesis, migration, immune escape and resistance to apoptosis. Galectin-1 (Gal-1) is a 14.5-kDa β-galactoside-binding protein, which displays intracellular (i.e., protein-protein interactions) and extracellular (i.e., protein-oligosaccharide interactions) functions with marked pro-angiogenic and pro-migratory effects in gliomas. Although glioma is considered one of the deadliest cancers with median survival ranging from nine to twelve months, therapeutic treatments that target Gal-1 may prove effective since Gal-1 is one of the genes that cause aggressive behaviors of human GBM such as migration and invasion. Gal-1 controls glioma cell migration and invasion by modifying the actin cytoskeleton and expression of small GTPases. In the present study, immunohistochemical, Real Time PCR and Western blot experiments performed with clinical samples demonstrated that Gal-1 is highly expressed in human GBM tissue as compared to healthy brain tissue. Matrigel invasion assays confirmed that treatment of glioma cell lines U251 and 5310 with human umbilical cord blood stem cells (hUCBSC) almost completely blocked invasion of GBM cells. We also observed that treatment of GBM cells with hUCBSC decreased the expression of both Gal-1 and FAK. Additionally, silencing either the Gal-1 or FAK gene using shRNA decreased migration and invasion of glioblastoma cells. Furthermore, using immunocytochemistry and Western blot analyses, we observed that Gal-1 and FAK interact with each other in glioblastoma. Taken together, our results suggest that Gal-1 and FAK might have a direct protein-protein interaction for their function in the tumorigenesis of glioblastoma.

Citation Format: Kiran Kumar Velpula, Arnima Bhasin, Jane S. Zhang, Andrew J. Tsung, Krishna Kumar Veeravalli, Jasti S. Rao, Venkata Ramesh Dasari. Galectin-1 modulates glioblastoma cell migration via interaction with FAK. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2635. doi:10.1158/1538-7445.AM2013-2635
Abstract 2640: HEF1 is an important regulator of mesenchymal and amoeboid invasion in breast cancer.
Authors: Jones Brandon , Pugacheva Elena
Published: August 2014

In previous studies, the focal adhesion-associated scaffolding protein HEF1, also known as NEDD9, has been implicated as a possible dynamic switch between the mesenchymal and amoeboid migration methods of melanoma cancer cells. Management of this switch provides a possible mechanism of control that could potentially open up new strategies for treatments of metastatic cancers. We have sought to investigate whether HEF1’s involvement in cell migration could also be applied to breast cancer. To monitor the switch between the two methods of migration, we decided to monitor activation of the Rac1 and RhoA proteins of the Rho subfamily of GTPases, which are renowned for their roles in actin cytoskeletal dynamics and have been linked to cancer cell invasion. High levels of active Rac1 result in an elongated cell morphology accompanied by mesenchymal cell movement, whereas high active RhoA levels result in a rounded cell morphology accompanied by amoeboid cell movement. Using a series of GST-tag pulldown experiments, we pulled and quantified amounts of active Rac1 and RhoA from an MDA-MB-231LN human breast cancer cell line grown in both 2D and 3D conditions under a HEF1 shRNA knockdown. Our HEF1-reduced cell line shows a decrease in active Rac1 and an increase in active RhoA compared to a control cell line with normal HEF1 levels. With this data, we aim to further substantiate HEF1 as a relevant key player in the balance between mesenchymal and amoeboid dominance in cancer cells.

Citation Format: Brandon Jones, Elena Pugacheva. HEF1 is an important regulator of mesenchymal and amoeboid invasion in breast cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2640. doi:10.1158/1538-7445.AM2013-2640
Abstract 2652: Multiplex detection of stem cell pluripotency markers in reference cancer cell lines.
Authors: Wiese Frederick W. , Stankiewicz Nikolai , Reiners Ralf , Warmke Tim , Maheshwari Reeti
Published: August 2014

Several proteins, including Oct-3/4, Sox2, KLF4 and Nanog, play a critical role in the maintenance of pluripotency in stem cells. Overexpression of one or a combination of these factors has been reported in solid tumors from a variety of tissues. The presence of pseudogenes and differentially expressed protein isoforms complicates the accurate detection of the physiologically relevant forms of these proteins in cancer stem cell research. Here, we report the development of MILLIPLEX® MAP multiplex assays for the simultaneous, specific and sensitive detection of the pluripotency markers Oct-3/4, Sox2, KLF4 and Nanog as well as several other stem cell relevant proteins in cellular lysates. Application of the assay to a panel of 25 human cancer cell lines from a variety of organs revealed Sox2 to be the most often expressed protein among these lines. Of the cell lines assayed, only one breast cancer cell line expressed detectable levels of all three transcription factors, suggesting a role for Oct3/4, Sox2 and Nanog in cancer stem cells of this particular cell line. These experiments demonstrate the utility of this pluripotency marker panel for the sensitive analysis of stem cells from a variety of lineages. The expression of the panel analytes was also examined in induced pluripotent stem cells (iPS) and a human embryonic stem cell (hES) line for reference. Taken together we show the successful development of a multiplex immunoassay that can be used to investigate the role of pluripotency marker proteins in stem cells or cancer cell immortalized or primary lines.

Citation Format: Frederick W. Wiese, Nikolai Stankiewicz, Ralf Reiners, Tim Warmke, Reeti Maheshwari. Multiplex detection of stem cell pluripotency markers in reference cancer cell lines. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2652. doi:10.1158/1538-7445.AM2013-2652
Abstract 2656: Using Nanog expression to identify breast cancer stem cell populations.
Authors: Brooks Alan D. , de Kluyver Rachel L. , Stauffer Jimmy K. , Kaddoura Marcella , Sayers Thomas J.
Published: August 2014

Many tumors have been demonstrated to contain a subpopulation of cancer stem cells (CSC) whose epigenetic modifications convey resistance to conventional therapies. Furthermore, tumor expression of genes associated with pluripotent embryonic cells, such as Oct-4, Sox-2 and Nanog, often correlates with poor prognosis. We hypothesized that expression of transcription factors that are required for induction and maintenance of pluripotency (Oct-4, Sox-2 and Nanog) would be a possible alternative to cell surface phenotype for identification of CSC populations. Therefore we evaluated Nanog expression in 4T1.2 murine breast carcinoma cells, as a biomarker for CSC sub-populations. Introduction of a Nanog promoter reporter that drives destabilized GFP into 4T1.2 cells resulted in a stable expression of GFP in about 1% of the cells. These GFP+ cell could be easily isolated by FACS cell sorting. On subsequent in vitro cell culture of GFP+ populations, up to 80% of the cells became GFP- over a period of 4 weeks. This would be consistent with a subsequent differentiation of these cells. By contrast GFP- populations remained 100% GFP-. In vitro CSC surrogate assays show that Nanog-GFP+ cells produced more spheroids in soft agar and under non-adherent growth conditions. More importantly, in vivo cell transfer studies demonstrated that Nanog-GFP+ cells were more efficient at generating experimental lung metastases when compared to Nanog-GFP-cells. Interestingly Nanog-GFP+ cells were more resistant to the chemotherapeutic drugs paclitaxel and bortezomib. However targeting the extrinsic apoptosis pathway with a combination of bortezomib and agonist antibodies to the TRAIL death receptor DR5 was equally effective against both Nanog-GFP+ and Nanog-GFP- cells. These findings suggest that Nanog promoter activity is a robust marker of highly metastatic 4T1.2 CSC subpopulations, and suggests that eradication of such cells may be required for improved anti-cancer therapies.

Funded by NCI Contract HHSN261200800001E

Citation Format: Alan D. Brooks, Rachel L. de Kluyver, Jimmy K. Stauffer, Marcella Kaddoura, Thomas J. Sayers. Using Nanog expression to identify breast cancer stem cell populations. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2656. doi:10.1158/1538-7445.AM2013-2656
Abstract 266: Notch3 is a marker of tumor-propagating cells in non-small cell lung cancer and is required for their self-renewal.
Authors: Zheng Yanyan , de la Cruz Cecile , Sayles Leanne , Alleyne-Chin Chris , Xu Yue , Hoang Chuong D. , Shrager Joseph B. , Jackson Erica , Sweet-Cordero Alejandro
Published: August 2014

We investigated tumor cell heterogeneity in mouse models of non-small cell lung cancer (NSCLC) that recapitulate key genetic events in this disease. In tumors driven by simultaneous expression of an oncogenic Kras allele and loss of the tumor suppressor Trp53, only a relatively rare subset of tumor cells is capable of propagating tumor growth in both a clonogenic in vitro assay and an in vivo orthotopic transplantation assay. The tumor-propagating cell (TPC) population was enriched by sorting for the combination of the cell surface markers CD24, ITGB4 and Notch(1-4). We demonstrate a critical and specific role for Notch3 signaling in the maintenance of TPCs in the KrasG12D; Trp53fl/fl mouse model. Knock-down of Notch3, but not Notch 1, 2 or 4, decreased self-renewal of TPCs in vitro. In addition, knock-down of Notch3 was critical for tumor propagation in vivo. The relevance of these studies to human NSCLC was confirmed by demonstrating a significant effect of Notch inhibition on the self-renewal of primary human lung adenocarcinoma cells in ex vivo cultures established directly from patients. These findings identify a novel subpopulation of tumor cells with distinct functional capabilities, underscoring the importance of interrogating heterogeneity in NSCLC. The unique role of Notch3 in tumor propagation may provide a novel therapeutic target for the treatment of non-small cell lung cancer.

 

Citation Format: Yanyan Zheng, Cecile de la Cruz, Leanne Sayles, Chris Alleyne-Chin, Yue Xu, Chuong D. Hoang, Joseph B. Shrager, Erica Jackson, Alejandro Sweet-Cordero. Notch3 is a marker of tumor-propagating cells in non-small cell lung cancer and is required for their self-renewal. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 266. doi:10.1158/1538-7445.AM2013-266
Abstract 2663: SPECT/CT analysis of distribution of intracranially infused interleukin-13 Pseudomonas exotoxin, a targeted agent for interleukin-13 receptor expressing brain tumors.
Authors: Suzuki Akiko , Leland Pamela , Choyke Peter L. , Inoue Tomio , Joshi Bharat H. , Puri Raj K.
Published: August 2014

Interleukin-13 Pseudomonas exotoxin (IL-13PE), a powerful targeted agent for IL-13 receptor α2 (IL-13Rα2) - positive tumors is being tested for safety and effectiveness in glioblastoma multiforme (GBM) clinical trials. However, distribution of IL-13PE after intra-cranial convection-enhanced delivery (CED) has not been determined. To study this, we radiolabeled IL-13PE with 125I and determined its binding and biological activity in vitro and distribution in a murine model of GBM. 125IL-13PE maintained its specific binding activity to IL-13Rα2 positive U251 glioblastoma tumor cells but not to T98G cells, as these cells did not express IL-13Rα2. 125IL-13PE was as cytotoxic to glioblastoma cells as non-radiolabeled IL-13PE, which killed U251 tumor cells in a concentration dependent manner but not T98G cells. Both binding and cytotoxic activities were blocked by a 100-fold excess of IL-13, which indicated that IL-13PE binding was specific. Athymic nude mice with intracranially implanted U251 tumors were given stereotactic intratumoral injections of 125I-IL-13PE by either a bolus injection or a 3-day-CED. Animals were imaged by micro-SPECT/CT at various time points. SPECT/CT imaging revealed retention of 125I-IL-13PE in U251 tumors. CED showed significantly higher volumes of distribution and maintained detectable drug levels for longer periods of time compared to bolus injection. SPECT/CT images of whole brains corroborated with the autoradiography results performed on sectioned brain. These results indicate that IL-13PE can be successfully radiolabled with 125I without the loss of binding and cytotoxic activity and that 125I-IL-13PE administered by CED showed higher volumes of distribution than the bolus route. Thus, future clinical studies with 125I-IL-13PE or other agents should consider the CED mode of delivery for better distribution in intracranial environments.

Volume of distribution of 125I-IL-13PE after bolus or CED administration into mouse brain Injection parameters SPECT Time Vi (μl)* Vd (mm3)** Vd:Vi ratio Bolus - day 1 10 minutes 5 10.2 ± 0.4 204.7 ± 7.0 Bolus - day 7 10 minutes 5 0.1 ± 0.02 2.0 ± 0.4 CED - day 1 3 days 100 120.4 ± 21.8 120.4 ± 21.6 CED - day 7 3 days 100 81.7 ± 6.9 81.7 ± 6.7 *Vi: Volume of injection **Vd: Volume of distribution

Citation Format: Akiko Suzuki, Pamela Leland, Peter L. Choyke, Tomio Inoue, Bharat H. Joshi, Raj K. Puri. SPECT/CT analysis of distribution of intracranially infused interleukin-13 Pseudomonas exotoxin, a targeted agent for interleukin-13 receptor expressing brain tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2663. doi:10.1158/1538-7445.AM2013-2663
Abstract 2692: Dendrimer delivery of RNA based Twist1 inhibitors to reduce breast cancer cell metastasis.
Authors: Finlay James B. , Roberts Cai M. , Glackin Carlotta A.
Published: August 2014

Breast cancer in women is a leading cause of cancer related deaths in the United States with metastatic disease being the chief cause of mortality. Metastatic breast cancer tumors are often found in their later stages making them difficult to remove or treat. Inhibition of tumor cell dispersion is therefore a key component to a reduction in mortality rates among women with breast cancer. Epithelial-Mesenchymal Transition (EMT) is the process by which cancer cells downregulate proteins associated with cell to cell adhesion (e.g. E-cadherin) resulting in cells that are able to mobilize. Breast cancer cells are known to commandeer the EMT process through overexpression of Twist1, thus allowing the cells to metastasize beyond the primary tumor. Twist1 therefore serves as an excellent therapeutic target whose disruption would result in fewer metastatic cancer cells and correspondingly reduce cancer mortality. Twist1 is also a desirable target because it is almost nonexistent in adult tissues and thus its silencing would have minimal side effects.

Our lab has elected to use an siRNA mediated gene silencing approach to decrease the expression levels of Twist1 in two highly invasive breast cancer cell lines (SUM 1315 and MCF7-Tw). Due to their fragile nature, siRNA molecules are often difficult to deliver at therapeutic levels. We have overcome these delivery limitations through the optimization and use of Poly (amidoamine) (PAMAM) dendrimers. We hypothesize that suppression of the activity of Twist1 via dendrimer-delivered Twist1 siRNA will inhibit metastatic behavior of aggressive breast cancer cells. Here we demonstrate successful delivery of Cy3-labled siRNA using two different dendrimers (Generation 5 and a modified Generation 3) with transfection efficiency results exceeding those of Lipofectamine 2000 and with far less toxicity to cells. We also show up to a 90% reduction (lasting at least 4 days) in Twist1 expression using Western Blot and qPCR analysis. Phenotypically we were able to appreciate a significant decrease in the invasive nature of the breast cancer cells using migration/invasion assays. Taken together these results demonstrate successful knockdown of Twist1 using siRNA dendrimer complexes resulting in an altered cellular phenotype. These data will serve as a foundation for optimization of future in vivo experiments with both orthotopic and metastatic breast cancer models.

Citation Format: James B. Finlay, Cai M. Roberts, Carlotta A. Glackin. Dendrimer delivery of RNA based Twist1 inhibitors to reduce breast cancer cell metastasis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2692. doi:10.1158/1538-7445.AM2013-2692
Abstract 2695: Osteopontin regulated epithelial-mesenchymal transition via PI3K/AKT signaling pathway in hepatocellular carcinoma.
Authors: Dong Qiongzhu , Zhu Xu-Chao , Dai Chun , Zhang Xiao-Fei , Jia Hu-Liang , Ye Qing-Hai , Qin Lun-Xiu
Published: August 2014

Our previous studies demonstrated that osteopontin (OPN) plays a crucial role in hepatocellular carcinoma (HCC) metastasis. However, its function in epithelial-mesenchymal transition (EMT) remains obscure. In this study, we reported the critical role of OPN in EMT mediated HCC metastasis. Increased expression of OPN in human HCC predicts poor survival and disease recurrence after surgery. HCC tumors overexpressing OPN were featured by active EMT and motility/invasion of tumor cells both in vitro and in vivo, whereas silencing OPN expression attenuated

EMT. OPN overexpression enhanced twist through activation of Akt, a critical EMT regulation of transcription factor. Suppression of the PI3K-AKT-Twist pathway interfered with OPN-mediated EMT and invasion. Consistently, we have observed a significant correlation between OPN expression and twist levels in a cohort of HCC patients, and found that combination of these two parameters is a more powerful predictor of poor prognosis.

Conclusions: OPN plays a pivotal role in EMT mediated HCC metastasis and may serve as a novel prognostic biomarker and therapeutic target.

Citation Format: Qiongzhu Dong, Xu-Chao Zhu, Chun Dai, Xiao-Fei Zhang, Hu-Liang Jia, Qing-Hai Ye, Lun-Xiu Qin. Osteopontin regulated epithelial-mesenchymal transition via PI3K/AKT signaling pathway in hepatocellular carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2695. doi:10.1158/1538-7445.AM2013-2695
Abstract 2697: Functional analysis of metastasis-related microRNA induced by the stromal-cell derived factor (SDF)-1/CXCR4 system in oral cancer.
Authors: Kinouchi Makoto , Uchida Daisuke , Kuribayashi Nobuyuki , Tamatani Tetsuya , Nagai Hirokazu , Miyamoto Youji
Published: August 2014

We have previously demonstrated that stromal cell-derived factor (SDF)-1/CXCR4 system is involved in the metastatic process of oral cancer. Recently, small non coding RNAs, microRNAs (miRNAs) are involved in the metastatic process of several types of cancers. However, the miRNAs contributed to metastases induced by the SDF-1/CXCR4 system in oral cancer are largely unknown. Thus, we examined the metastasis-related miRNAs induced by the SDF-1/CXCR4 system, using oral cancer cells, B88-SDF-1, which exhibits functional CXCR4 and distant metastatic potential, in vivo. We examined the metastasis-related miRNAs induced by the SDF-1/CXCR4 system by use of a miRNA microarray representing approximately 1700 miRNA. Consequnetly, we identified 4 kinds of upregulated-miRNAs in B88-SDF-1 cells whose functions are unknown. Among these miRNAs, we confirmed the upregulation of miR-5XX in B88-SDF-1 cells by the real-time PCR analysis. Although LNA-based miR-5XX inhibitor did not affect the growth of the cells, it did significantly inhibit the migration of B88-SDF-1 cells. Thus, we examined the target gene(s) of miR-5XX in the SDF-1/CXCR4 system. cDNA microarray analysis revealed that 6 kinds of mRNA containing the predictive binding site of miR-5XX were upregulated by the treatment with miR-5XX inhibitor. Furthermore, gene A mRNA and protein was downregulated in B88-SDF-1 cells, and upregulated by the treatment with miR-5XX inhibitor. These results indicating that SDF-1/CXCR4 system might regulate the metastases of oral cancer by the downregulation of gene A via upregulation of miR-5XX.

Citation Format: Makoto Kinouchi, Daisuke Uchida, Nobuyuki Kuribayashi, Tetsuya Tamatani, Hirokazu Nagai, Youji Miyamoto. Functional analysis of metastasis-related microRNA induced by the stromal-cell derived factor (SDF)-1/CXCR4 system in oral cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2697. doi:10.1158/1538-7445.AM2013-2697
Abstract 27: A phase I study of MEK inhibitor BAY 86-9766 in Japanese patients with advanced or reflactory solid tumors.
Authors: Doi Toshihiko , Fuse Nozomi , Yoshino Takayuki , Shitara Kohei , Bando Hideaki , Miyamoto Hideaki , Koyama Susumu , Tsukada Katsuya , Ohtsu Atsushi
Published: August 2014

Background: BAY 86-9766 (RDEA119) is a potent, orally administered,, allosteric inhibitor of mitogen-activated protein (MAP) kinase kinase (MEK) 1/2, a key protein in the MAP kinase signaling pathway, located downstream of RAS and RAF. We conducted a phase I study to evaluate the safety and pharmacokinetics (PK) of BAY 86-9766 in advanced/refractory Japanese cancer patients.

Patients and Methods: BAY 86-9766 was administered at 30 mg bid (cohort 1) and at 50 mg bid (MTD; cohort 2) over a 28-day treatment cycle which was continued until disease progression or unacceptable toxicity. PK samples were collected after single doses (Day 1 to Day 4) and during multiple dosing through Day 22. Tumor response was assessed after every even cycle by RECIST v.1.1.

Results: A total of 12 patients were enrolled, including 6 colorectal cancer, 2 GIST, and one hepatocellular cancer, carcinoid tumor, neuroendocrine tumor and leiomyosarcoma, respectively (5 patients in cohort 1: 30mg bid; 7 patients in cohort 2: 50mg bid). There were no DLTs in Cycle 1 of cohort 1 and cohort 2. The most common adverse event regardless of grade was rash acneiform in 12 subjects (100.0%), followed by elevated AST, creatine phosphokinase (CPK) or hypoalbuminemia in 9 subjects (75.0%), fever or elevated ALP in 7 subjects (58.3%), elevated ALT or decreased platelet count in 6 subjects (50.0%). Of those, the most common drug-related adverse events were rash acneiform (100.0%), followed by elevated CPK (66.7%), laboratory-others (66.7%), elevated AST (58.3%), and elevated ALP (50.0%). There were three cases of serious adverse events - thrombocytopenia, bile duct stenosis and elevated CPK - with only the elevated CPK being regarded as drug-related. There were no deaths on study. In Cohort 2, 7 out of 7 patients had elevated CPK, with 4 patients Grade 3 or higher. None of these were considered clinically significant and were not regarded as DLT's. Three patients had a positive de-challenge and re-challenge and 2 patients experienced muscle-related symptoms (neck pain and muscle weakness of the lower limb). PK parameters were comparable to the non-Japanese population.

Conclusions: BAY 86-9766 was generally well tolerated at both the 30 mg bid dose and the MTD of 50 mg bid. BAY 86-9766 administration increased CPK in a majority of patients but this was manageable by dose reduction.

Citation Format: Toshihiko Doi, Nozomi Fuse, Takayuki Yoshino, Kohei Shitara, Hideaki Bando, Hideaki Miyamoto, Susumu Koyama, Katsuya Tsukada, Atsushi Ohtsu. A phase I study of MEK inhibitor BAY 86-9766 in Japanese patients with advanced or reflactory solid tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 27. doi:10.1158/1538-7445.AM2013-27
Abstract 2701: Expression and function of CXCR4 in human salivary gland cancers.
Authors: Kuribayashi Nobuyuki , Uchida Daisuke , Kudo Yasusei , Kinouchi Makoto , Tamatani Tetsuya , Nagai Hirokazu , Miyamoto Youji
Published: August 2014

Salivary gland cancers (SGCs) frequently metastasize to cervical lymph nodes and distant organs. Currently, the mechanisms responsible for the metastatic behavior of SGC cells are not fully understood. We previously demonstrated that the stromal cell-derived factor-1 (SDF-1; also known as CXCL12)/CXCR4 system is involved in the establishment of metastasis in oral squamous cell carcinoma. In the present study, we investigated the role of CXCR4 in the metastatic behavior of SGCs. We examined the expression of CXCR4 mRNA and protein in human SGC cell lines by quantitative RT-PCR and western blotting, respectively. The expression of CXCR4 mRNA and protein were frequently upregulated in 5 out of 6 SGC cell lines. On the contrary to the CXCR4 expression, CXCR7, a recently identified second receptor for SDF-1, was detected in all SGC cell lines, expression levels were considerably lower on average compared with that of CXCR4 expression. Functional CXCR4 expression was demonstrated by the ability of these SGC cell lines to migrate toward an SDF-1 gradient. SDF-1 rapidly activated extracellular signal-regulated kinase (ERK)1/2 in SGC cell lines. Immunohistochemical analysis revealed that CXCR4 protein expression was detected in either the nucleus or cytoplasm of cancer cells in 16 out of 20 tissues of adenoid cystic carcinoma (ACC) and in 4 out of 6 tissues of mucoepidermoid carcinoma, which are representative of SGC. Furthermore, ACC cell lines exhibited dramatic metastasis to the lung following intravenous inoculation, whereas AMD3100, a CXCR4 antagonist, significantly inhibited lung metastasis of the cells, ameliorated body weight loss and improved the survival rate of tumor-bearing nude mice. These results indicate that CXCR4 expression contributes to the metastatic potential of SGCs.

Citation Format: Nobuyuki Kuribayashi, Daisuke Uchida, Yasusei Kudo, Makoto Kinouchi, Tetsuya Tamatani, Hirokazu Nagai, Youji Miyamoto. Expression and function of CXCR4 in human salivary gland cancers. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2701. doi:10.1158/1538-7445.AM2013-2701