bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3464: The effect of EPO-receptor in estrogen receptor positive breast cancer
Authors: Vaapil Marica C. , Reinbothe Susann , Larsson Anna-Maria , Wigerup Caroline , Sun Jianmin , Jögi Annika , Neumann Drorit , Rönnstrand Lars , Påhlman Sven
Published: May 2015

The main function of Erythropoietin (EPO) and its receptor (EPOR) is the stimulation of erythropoiesis. Recombinant human EPO (rhEPO) is therefore used to treat anemia in cancer patients. However, recent clinical trials have indicated that rhEPO treatment might promote tumor progression and have a negative effect on patient survival. In addition, EPOR expression has been detected in several cancer forms. Using a newly produced anti-EPOR antibody that reliably detects the full-length isoform of the EPOR we show that breast cancer tissue and cells express the EPOR protein. rhEPO stimulation of cultured EPOR expressing breast cancer cells did not result in increased proliferation, overt activation of EPOR (receptor phosphorylation) or a consistent activation of canonical EPOR signaling pathway mediators such as JAK2, STAT3, STAT5, or AKT. However, EPOR knockdown experiments suggested functional EPO receptors in estrogen receptor positive (ERα+) breast cancer cells, as reduced EPOR expression resulted in decreased proliferation. This effect on proliferation was not seen in ERα negative cells. EPOR knockdown decreased ERα activity in ERα+ breast cancer cells and diminished ERα+ phosphorylation on Ser118, results that further support a mechanism by which EPOR affects proliferation via ERα+ mediated mechanisms. In conclusion, we show that EPOR protein is expressed in breast cancer cells, where it appears to promote proliferation in ERα+ expressing breast cancer cells, in part via its capacity to positively modulate ERα activity.

Citation Format: Marica C. Vaapil, Susann Reinbothe, Anna-Maria Larsson, Caroline Wigerup, Jianmin Sun, Annika Jögi, Drorit Neumann, Lars Rönnstrand, Sven Påhlman. The effect of EPO-receptor in estrogen receptor positive breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3464. doi:10.1158/1538-7445.AM2014-3464
Abstract 3468: Integrin-β6 is required for transforming growth factor (TGF)-β-mediated epithelial-mesenchymal transition (EMT) and promotes the disease progression of intrahepatic cholangiocellular carcinoma
Authors: Higashi Takaaki , Hayashi Hiromitu , Kuroki Hideyuki , Nakagawa Shigeki , Sakamoto Keita , Nitta Hidetoshi , Chikamoto Akira , Toru Toru , Baba Hideo
Published: May 2015

Background: Intrahepatic cholangiocellular carcinoma (ICC) is characterized as a poor prognostic disease represented by the highly invasiveness. Epithelial-mesenchymal transition (EMT) plays a pivotal role in cancer invasion and metastasis, and then TGF-β signaling is a potent inducible factor of EMT. Integrin-β6, which is an epithelial restricted trans-membrane protein, promotes binding to extracellular matrix proteins (fibronectin, vitronectin and tenascin) facilitating cell migration and activates TGFβ1 via binding to the latency associated peptide (LAP) of the TGF-β complex during embryogenesis and wound healing. In this study, we investigated the role of integrin-β6 during TGF-β-mediated EMT using ICC cell lines, and examined the clinic-pathological significance of Integrinβ6 expression in ICC patients.

Material and Methods: We measured integrin-β6 mRNA expression by real time PCR using 7 ICC cell lines (Hucct1, HuH28, OZ, RBE, Ssp25, TFK1, Ysccc). To examine the role of integrin-β6 during TGF-β-mediated EMT in ICC cell lines, we downregulated integrin-β6 expression by RNAi during TGF-β-medaited EMT. Furthermore, using 55 resected ICC specimens, we evaluated integrin-β6 expression by immunohistochemical staining.

Results: Of 7 ICC cell lines, Ysccc and OZ cell lines revealed high integrin-β6 expressions. Addition of TGF-β to Ysccc cell successfully induced EMT, characterized by the upregulated vimentin expression, downregulated E-cadherin expression, spindle-shaped morphology, and high invasiveness, with the integrin-β6 induction. The inhibition of integrin-β6 expression using RNAi abolished TGF-β-mediated EMT induction. In clinical setting, high integrin-β6 expression was significantly associated with Stage IV, lymph node metastasis, and blood vessel invasion, and then resulted in worse prognosis compared with low expression group.

Conclusion : Integrin-β6 is required for TGF-β-mediated EMT and promotes the disease progression of ICC.

Citation Format: Takaaki Higashi, Hiromitu Hayashi, Hideyuki Kuroki, Shigeki Nakagawa, Keita Sakamoto, Hidetoshi Nitta, Akira Chikamoto, Toru Toru, Hideo Baba. Integrin-β6 is required for transforming growth factor (TGF)-β-mediated epithelial-mesenchymal transition (EMT) and promotes the disease progression of intrahepatic cholangiocellular carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3468. doi:10.1158/1538-7445.AM2014-3468
Abstract 3469: The effect of TGF-beta on matrix metalloproteinase-2, 9, and 13 expression and secrection in human prostate cancer cells
Authors: Cao Yang , Bolton Clement J. , Khan Shafiq A.
Published: May 2015

Matrix metalloproteinases (MMPs) are major critical molecules assisting tumor cells during metastasis. The production of MMPs by prostatic epithelial and/or neighboring stromal cells plays an important role in the penetration of extracellular matrix barriers in normal or neoplastic growth. Activation of MMP-13 is critical in the MMP cascade and MMP-2 and -9 are important in tumor metastasis. Transforming growth factor (TGF)-beta has been reported to exert growth inhibitory activity in normal epithelial cells whereas it induces cell proliferation and invasive phenotypes in advanced carcinomas.

In this study, we investigated the role of TGF-β1 and TGF-β3 in MMP expression and secretion in prostate cancer cell line PC-3 and prostatic epithelial cell line RWPE-1. Treatment with 1ng/ml of TGF-β1 and TGF-β3 enhanced MMP-9 expression in both cell lines but showed no significant effect on MMP-2 expression. TGF-β1 and TGF-β3 increased MMP-13 expression in PC-3 cells at 8 hr but decreased its expression in RWPE-1 cells. As examined in conditioned medium (CM) by zymography, TGF-β1 and TGF-β3 increased the secretion of pro-MMP-9 and its active form at different time points in PC-3cells. However, they inhibited pro-MMP-2 and no active MMP-2 activity was detected in the same CM. On the other hand, both TGF-β1 and TGF-β3 increased pro-MMP-9 and pro-MMP-2 in RWPE-1 cells. TGF-β3 is more potent that TGF-β1. TGF-β3 also increased active MMP-2 after 18 hr. MMP-9 siRNA abolished TGF-β induced invasion in PC-3 cells but MMP-13 siRNA didn’t affect PC3 cell migration. Our results suggest that TGF-β1 and TGF-β3 have stimulatory effects on MMP-9 secretion in both PC-3 and RWPE-1cells and similar effect on MMP-2 secretion in RWPE-1 cells. However, they have inhibitory effect on MMP-2 secretion in PC-3 cells. MMP-9 is essential for TGF-β effects on cell migration. TGF-β has opposite effect on MMP-13 expression in the two cell lines and MMP-13 is not required for cell migration.

Citation Format: Yang Cao, Clement J. Bolton, Shafiq A. Khan. The effect of TGF-beta on matrix metalloproteinase-2, 9, and 13 expression and secrection in human prostate cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3469. doi:10.1158/1538-7445.AM2014-3469
Abstract 347: Dasatinib accelerates valproic acid-induced acute myeloid leukemia cells death by regulation of differentiation capacity
Authors: Heo Sook-Kyoung , Noh Eui-Kyu , Yoon Dong-Joon , Jo Jae-Cheol , Park Jae-Hoo , Kim Hawk
Published: May 2015

Dastinib (BMS-354825) is a FDA-approved small compound that was developed for chronic myeloid leukemia as a multi-targeted kinase inhibitor of wild-type BCR-ABL and SRC family kinases. Valproic acid (VPA; Orfiril) has been a well-known anti-epileptic drug worldwidely, and belong to the class I histone deacetylase inhibitor. The aim of this research is to determine the anti-leukemic effects of combination of dasatinib and VPA or their mechanism in acute myeloid leukemia (AML) cells. In this study, dasatinib showed potent inhibitory effects on VPA-treated AML cells that were associated with induced G1 phase cell cycle arrest and also apoptosis including the cleavage of PARP, caspase-3, -7 and -9. Moreover, inhibitors of caspase-3 and -9 (Z-DEVD-FMK and LEHD-CHO) efficiently inhibited dasatinib-induced apoptosis in VPA-treated AML cells. Therefore, dasatinib/VPA-induced cell death occurred via caspase-dependent apoptosis. Also, inhibitors of MEK/ERK (U0126 and PD98059) and p38 MAPK (SB203580) efficiently inhibited dasatinib/VPA-induced apoptosis. The combination of dasatinib and VPA differentiation capacity of AML cells is more powerful than each drug alone. So the combination is enough to push AML cells to death strongly by G1 cell cycle arrest and caspase-dependent apoptosis. In addition, MEK/ERK and p38 MAPK controls dasatinib/VPA-induced apoptosis as an upstream regulator. Finally, co-treatment of dasatinib and VPA is contributed to AML cell death by regulation of differentiation capacity. Taken together, these results indicate that dasatinib and VPA together have a potential for anti-leukemic therapy.

Citation Format: Sook-Kyoung Heo, Eui-Kyu Noh, Dong-Joon Yoon, Jae-Cheol Jo, Jae-Hoo Park, Hawk Kim. Dasatinib accelerates valproic acid-induced acute myeloid leukemia cells death by regulation of differentiation capacity. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 347. doi:10.1158/1538-7445.AM2014-347
Abstract 3475: Negative impact of evaporation on cervical cancer cells during long term incubation is alleviated by Thermo Scientific Edge plate
Authors: Neeley Cindy , Verna Sarah
Published: May 2015

With more than half a million new cases and 275,000 deaths each year, cervical cancer ranks as the third and the fourth most common cause of cancer incidence and mortality, respectively, in women worldwide. Prophylactics, screening, testing, and treatment options for cervical cancer lead to an increasing need for new and innovative technologies. Microplates are widely engaged in all aspects of cervical cancer research. However, researchers have long struggled with media evaporation from the perimeter wells of their microplates. This condition negatively affects research results and lab productivity. In this study, we demonstrated that human cervical cancer HeLa cells suffered significant reduction in cell viability over 7-day incubation in the perimeter wells of standard 96-well plate. HeLa cells in these wells underwent premature NFκB nuclear translocation suggesting the onset of cell stress during culture. In comparison, study using the Thermo Scientific Edge plate with an innovative design of “build-in” mote showed significantly less evaporation from the perimeter wells. The variations between perimeter wells and center wells were minimal in Edge plate, indicating uniformed performance across the entire plate. Advantages of Edge plate were further demonstrated by the improvement in HeLa cell viability in perimeter wells when compared to standard 96-well plate. HeLa cells cultured in Edge plate were in a much healthier state and only demonstrated NFκB nuclear translocation when stress-inducing agent TNF-α was applied to the culture. In summary, Edge plate allows the use of all 96 wells without the concern of evaporation, resulting in improved consistency in assay models for in vitro research of cervical and other types of cancer.

Citation Format: Cindy Neeley, Sarah Verna. Negative impact of evaporation on cervical cancer cells during long term incubation is alleviated by Thermo Scientific Edge plate. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3475. doi:10.1158/1538-7445.AM2014-3475
Abstract 3479: The cellular thermal shift assay as a tool for setting indivual treatment schemes
Authors: Molina Daniel M. , Nordlund Pär
Published: May 2015

Many cancers respond insufficiently to existing therapies and acquired drug resistance often develops. To address this challenge, a range of different therapeutic strategies is currently in development. Many of these are based on the concept of targeted therapies, where target proteins are drivers of cancer processes or synthetic vulnerabilities of cancer cells. In this project we have used the newly developed CETSA method to monitor levels of drug binding to specific target proteins in tumors and primary cells from breast cancer patients. CETSA is a generic method for evaluating drug binding to target proteins in cells and tissues. The technique is based on the physical phenomenon of ligand-induced thermal stabilization of target proteins. Using this technique, it is possible to look at interactions between a drug and protein of interest without having to label them. Previously, we have validated drug binding in mammalian cancer cell lines for a set of important clinical targets and shown that a range of critical factors that are important for drug development can be identified at the target engagement level, including drug transport and activation, off-target effects, drug resistance as well as drug distribution in animal tissues.

By using different drugs and amounts we now have preliminary data suggesting that CETSA could be used as a tool for optimizing treatment schemes for individual care and for detecting resistance development at an early stage.

Citation Format: Daniel M. Molina, Pär Nordlund. The cellular thermal shift assay as a tool for setting indivual treatment schemes. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3479. doi:10.1158/1538-7445.AM2014-3479
Abstract 3481: Genetically-engineered mice as a cell line isolating tool
Authors: Kamiyama Hirohiko , Rauenzahn Sherri , Maitra Anirban , Eshleman James R.
Published: May 2015

BACKGROUND

Isolation of cell lines from patients’ tumors is difficult, due in large part, to contaminating fibroblasts that often overgrow the malignant cells. The same is true of xenografted tumors. To overcome this problem, we developed biochemically selectable mouse cells by using genetically-engineered mice.

METHODS

We constructed a series of hypoxanthine phosphoribosyl transferase (hprt) null immunodeficient mice. These include nude hprt-null, SCID hprt-null and NOD-SCID hprt-null mice.

Tumors from patients were implanted into these mice for growth. During growth of xenografted human cancers, hprt-null murine stromal cells replace their human counterparts to facilitate the growth of human cancer cells. Explanted tumors were then grown in tissue culture flasks with selection media (hypoxanthine, aminopterin, thymine; HAT media) to eliminate the mouse biochemically-defective fibroblasts and produce pure human cancer cell lines.

RESULTS

We successfully established several pancreatic cancer cell lines and an ovarian cancer cell line from patients using the hprt null immunodeficient mice.

CONCLUSIONS

The new cell line isolating system using genetically-engineered mice provides a greater chance to isolate pure cancer cell lines from patients’ tumors. This may allow personalized chemotherapy selection.

Citation Format: Hirohiko Kamiyama, Sherri Rauenzahn, Anirban Maitra, James R. Eshleman. Genetically-engineered mice as a cell line isolating tool. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3481. doi:10.1158/1538-7445.AM2014-3481
Abstract 3488: HPV detection by PCR in real time in peruvian women
Authors: Manrique Javier , Sarria Gustavo , Arias Abelardo , Mora Pamela , Limache Abel , Nuñez Maria del Carmen , Sullcahuaman Yasser , Vidaurre Tatiana
Published: May 2015

Background. In Perú Cervical cancer is a significant public health problem, according Globocan, it is the most frequent cancer among women in Peru, the incidence of this cancer is 34.5 and the mortality 16.3. In 2004, cervical cancer was responsible of 66,771 DALYs in Peru, 18.2% of the total. The human papillomavirus (HPV) is a well-known cause of cervical cancer. HPV tests are used as an adjunct test to decrease the false-negative rate of cytological screening. The HPV test will permit to separate the population in two groups, women with high risk to cancer cervical (HPV+) and women with low risk to cancer cervical (HPV-).

Objective. To identify the prevalence of the genotypes of Human Papillomavirus (HPV) in Peruvian women from marginal urban area of Lima according age.

Methodology. Samples of 5460 women aged 18 to 63 years were studied for analysis by the automated Roche Cobas 4800 system that detects genotypes 16 and 18 specifically and 12 other genotypes (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 66) considered high risk HPV, for this purpose the system uses specific primers for the L1 region of the viral genome and control the system using B globin gene.

Results and Conclusions. According PCR screening the total samples were 5,460 (100%), 855 (15.62%) were positive by PCR, of the PCR positive 319 were positive by CBL (5.84%), 16 of them were LIE-AG (0.29%) and 0 carcinomas. The genotypes of PVH more frequent was 16 (16%), genotype 16 plus others high risk virus was 69 (8%), PVH 18 was 8 (3%). Of the 853 women that were positive for PCR, 149 of them had the highest risk HPV genotype 16 and the age group that most often present for this genotype was 30 to 39 years, this group is consider as high risk population. In general, PCR have been demonstrated an adequate prevalence rate of women in high risk according of other literature and we consider as an important test to cervical cancer control.

Ilustración 1. Frequency of HPV in Peruvian women.

Illustration 2. HPV Genotypes in peruvian women

Illustration 3. Distribution of CBL (+)

Ilustración 3. Genotipos de PVH y distribución por grupo etáreo. / HPV genotype distribution per age group.

Citation Format: Javier Manrique, Gustavo Sarria, Abelardo Arias, Pamela Mora, Abel Limache, Maria del Carmen Nuñez, Yasser Sullcahuaman, Tatiana Vidaurre. HPV detection by PCR in real time in peruvian women. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3488. doi:10.1158/1538-7445.AM2014-3488
Abstract 3495: A nucleolus-predominant piggyBac transposase increases transposition efficiency in human cancer cells
Authors: Hong Jin-Bon , Chou Fu-Ju , Ku Amy T. , Fan Hsiang-Hsuan , Lee Tung-Lung , Huang Yung-Hsin , Su I-Chang , Chen You-Tzung
Published: May 2015

PiggyBac is a prevalent transposon system used to deliver transgenes into and to functionally explore the mammalian untouched genomic territory. Efforts to increase its transposition efficiency in mammals through engineering the corresponding transposase (PBase) codon usage to improve its expression level is reported. The hyperactive PBase developed through screening for mutant variants are also established. To increase the safety for its potential use in regenerative medicine applications, site-specific transposition using engineered Zn finger-, Gal4- fused PBase, and an excision-prone PBase variant are also successfully developed. Here we report the construction of a nucleolus predominant PBase, NP-mPB, by adding a nucleolus predominant (NP) signal peptide from HIV-1 TAT protein to a mammalian codon optimized PBase (mPB). Although there is a predominant fraction of the NP-mPB-tGFP fusion proteins concentrated in the nucleoli, an insertion site preference toward nuleolar organizer regions (NORs) is not detected. Instead a 3-fold increase in piggyBac transposition efficiency is found in human cancer cell line.

Citation Format: Jin-Bon Hong, Fu-Ju Chou, Amy T. Ku, Hsiang-Hsuan Fan, Tung-Lung Lee, Yung-Hsin Huang, I-Chang Su, You-Tzung Chen. A nucleolus-predominant piggyBac transposase increases transposition efficiency in human cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3495. doi:10.1158/1538-7445.AM2014-3495
Abstract 3500: NCI's Provocative Questions Initiative
Authors: Berny-Lang Michelle A. , Greenspan Emily J.
Published: May 2015

In 2011, the NCI Director, Dr. Harold Varmus, created the Provocative Questions (PQ) initiative to encourage imaginative, bold approaches designed to tackle perplexing and previously neglected areas of cancer research. Based upon workshops and discussions with the extramural cancer research community and NCI program directors, a set of 24 questions and accompanying requests for applications (RFAs) were developed, addressing diverse topics in cancer risk and prevention, tumor development, and cancer detection, diagnosis, and treatment. In order to stimulate innovative hypotheses and approaches, the “power of the ideas” was heavily weighted with a de-emphasis on preliminary data. In the first year of the initiative, the NCI awarded 56 unique proposals, totaling $21.5 million of research support via the R01 and R21 mechanisms. Building upon the 2011 RFAs and adjusting to changes in the research landscape, 24 revised questions were developed for the 2012 RFAs with a commitment of up to $30 million in support. The NCI released the 2013 RFAs in September 2013 with a revised set of 20 questions, including new questions related to clinical effectiveness. Several PQ-supported research projects funded in the first year of the initiative will be highlighted in order to demonstrate the breadth of topics and high level of innovation supported by the initiative. Projects will be selected from the four main PQ thematic areas to provide a sense of the many diverse approaches being used to answer the selected PQ. In addition, trends and criteria from awarded proposals will be presented to provide insight to potential applicants regarding the outcomes of the program to date. Through engagement of the cancer research community, the PQ initiative seeks to continually leverage scientific advances to address currently unsolved, understudied questions in cancer, which may influence the future directions of NCI-sponsored research.

Citation Format: Michelle A. Berny-Lang, Emily J. Greenspan. NCI's Provocative Questions Initiative. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3500. doi:10.1158/1538-7445.AM2014-3500
Abstract 3501: Aspirin modulates oncogene expression in hct 116 colon cancer cells
Authors: Dachineni Rakesh , Ai Goqiang , Gunaje Jayarama B.
Published: May 2015

Aspirin is a 100 year old Non-steroidal anti-inflammatory drug (NSAID). Beside its use a cardio-protective agent, it is also used for treatment of headache, fever, and inflammation. Many studies have shown that regular intake of aspirin for 5 years or more reduces the risk of colon, breast, prostate, lung and skin cancers. However, mechanism for such actions is not very well understood. Since transformation of a normal cell to cancer cell often involves mutation and activation of proto-oncogene, we initiated this research to understand if aspirin modulates the level of oncogene products. In this study we focused on three different oncogenes; c-FOS, c-JUN and c-RAF. c-FOS and c-JUN are transcription factors, they form a heterodimer called Activator protein (AP-1) transcription factor and regulate the expression of other genes. We observed that, in serum starved HCT-116 colon cancer cells Aspirin and Salicylic acid inhibited serum induced c-FOS protein expression, however, levels of c-JUN protein were not inhibited. In fact Aspirin and Salicylic acid increased c-JUN levels marginally over the serum induced levels at all time points examined. Exposure of cells to Aspirin and Salicylic acid inhibited c-RAF levels in a concentration dependent fashion. Inhibition was observed at physiologically achieved concentration of 250 -500 micro molar Salicylic acid suggesting that Aspirin may mediate its effects through the formation of Salicylic acid. We are currently investigating how inhibition of c-RAF levels by Aspirin and Salicylic acid affects downstream signaling molecules such as p42/p44 MAP kinases. Modulation of proto oncogenes by Aspirin may represent an important pathway by which Aspirin may mediate its anti-cancer effects.

Citation Format: Rakesh Dachineni, Goqiang Ai, Jayarama B. Gunaje. Aspirin modulates oncogene expression in hct 116 colon cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3501. doi:10.1158/1538-7445.AM2014-3501
Abstract 3507: STAT3beta suppresses tumorigenesis via modulating the phosphorylation dynamics and transcription activity of STAT3alpha in esophageal squamous cell carcinoma
Authors: Zhang Haifeng , Lai Raymond , Li Enmin , Xu Liyan
Published: May 2015

While the oncogenic role of STAT3 is well-documented, a number of studies have described that STAT3 carries tumor suppressor functions. We hypothesized that the relative abundance of STAT3α and STAT3β, the two STAT3 isoforms, dictate its exact functional roles in cancer cells. Using Western blots and a cohort of esophageal squamous cell carcinoma (ESCC)(n=91), we found that STAT3β significantly correlated with a shorter survival (P=0.030), whereas phospho-STAT3α-Tyr705 and total STAT3 did not carry significant prognostic value. Correlating with these observations, STAT3β significantly suppressed the oncogenic effects of STAT3α in ESCC cell lines in a soft-agar clonogenic assay. Interestingly, while STAT3β suppressed the transcriptional activity of STAT3α, it enhanced the phosphorylation of STAT3α-Tyr705 and its nuclear translocation. STAT3β also prolonged the phosphorylation of STAT3α-Tyr705 and increased its cytoplasmic localization following cytokine stimulation. Furthermore, the substantial increase in phospho-STAT3α-Tyr705 in this setting was related to STAT3β-mediated inhibition of dephosphorylation. Taken together, our findings have highlighted the tumor suppressor roles of STAT3β via its interactions with STAT3α at different levels. Our data also strongly suggests that, an accurate interpretation of the oncogenic activity of STAT3α in tumors based on the detection of nuclear STAT3 or phospho-STAT3-Tyr705 requires the knowledge of the STAT3β expression status.

Citation Format: Haifeng Zhang, Raymond Lai, Enmin Li, Liyan Xu. STAT3beta suppresses tumorigenesis via modulating the phosphorylation dynamics and transcription activity of STAT3alpha in esophageal squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3507. doi:10.1158/1538-7445.AM2014-3507
Abstract 3512: MYCN and survivin cooperatively contribute to malignant transformation of fibroblasts
Authors: Hipp Nora , Christner Lisa , Wirth Thomas , Mueller-Klieser Wolfgang , Walenta Stefan , Schröck Evelin , Debatin Klaus-Michael , Beltinger Christian
Published: May 2015

The oncogenes MYCN and survivin (BIRC5) maintain aggressiveness of diverse cancers including sarcomas. To investigate whether these oncogenes cooperate in initial malignant transformation, we transduced them into Rat-1 fibroblasts. Indeed, survivin enhanced MYCN-driven contact-uninhibited and anchorage-independent growth in vitro. Importantly, upon subcutaneous transplantation into mice, cells overexpressing both instead of either one of the oncogenes generated tumors with shortened latency, marked anaplasia and an increased proliferation-to-apoptosis ratio resulting in accelerated growth. Mechanistically, the increased tumorigenicity was associated with an enhanced Warburg effect and HIF1α-linked vascular remodeling. This cooperation between MYCN and survivin may be important in the genesis of several cancers.

Citation Format: Nora Hipp, Lisa Christner, Thomas Wirth, Wolfgang Mueller-Klieser, Stefan Walenta, Evelin Schröck, Klaus-Michael Debatin, Christian Beltinger. MYCN and survivin cooperatively contribute to malignant transformation of fibroblasts. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3512. doi:10.1158/1538-7445.AM2014-3512
Abstract 3516: The mitochondrial sirtuin SIRT3 promotes survival of human melanoma cells in vitro
Authors: George Jasmine , Nihal Minakshi , Singh Chandra K. , Zhong Weixiong , Ahmad Nihal
Published: May 2015

Melanoma, a potentially lethal form of skin cancer, is the fifth most common type of new cancer diagnosis in American men and the sixth most common type in American women. Alarmingly, the annual incidence of melanoma among whites has increased steadily (>60% over the past 30 years). Despite new developments in therapy, the overall survival of patients with advance-stage melanoma has not significantly improved. Therefore, concerted mechanism-based efforts are needed for prevention, diagnosis and treatment of this deadly neoplasm. A better understanding of molecular mechanisms contributing to melanoma development and progression can be useful in crafting new strategies for melanoma management. We are studying the role of sirtuin family of proteins in melanoma development and progression. Mammalian sirtuins (SIRT 1-7) are nicotinamide adenine dinucleotide (NAD+)-dependent protein deacetylases, which have been shown to regulate metabolism, stress responses, and longevity, in model organisms. The role of sirtuins in cancer is fairly complex with dichotomous functions (tumor promoter versus tumor suppressor) depending on cell contexts. SIRT3 is a mitochondrial deacetylase that has been shown to be important in maintaining cellular metabolism by regulating mitochondrial functions. We have previously shown that SIRT3 was overexpressed in human melanoma cells and tissues and its inhibition resulted in an antiproliferative response in human melanoma cells (In: AACR Annual Meeting: Proceedings; 2013 April 6-April 10, Washington, DC; Abstract # 13-LB-9391). In this study, we found that short hairpin RNA (shRNA)-mediated knockdown of SIRT3 in SK-MEL-28 melanoma cells resulted in 1) G1-phase arrest of cell cycle, 2) induction of senescence as assessed by increase in senescence-associated β-galactosidase (SA-β-gal) activity, 3) senescence associated hetrochromatin formation (SAHF), 4) decrease in cell migration, and 5) increase in protein levels of p53, phospho-p53 (Ser15), p21Waf1, pRb and p16INK4a. Further, a stable exogenous overexpression of SIRT3 resulted in an increase in cell growth and clonogenic survival in Hs 294T melanoma cells. Based on our data we suggest that SIRT3 could be a contributing factor in melanoma survival. However, further extensive in-vitro and in- vivo studies are needed to ascertain the functional significance of SIRT3 in melanoma development and progression.

Citation Format: Jasmine George, Minakshi Nihal, Chandra K. Singh, Weixiong Zhong, Nihal Ahmad. The mitochondrial sirtuin SIRT3 promotes survival of human melanoma cells in vitro. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3516. doi:10.1158/1538-7445.AM2014-3516
Abstract 3525: SOX2 promotes tumor growth via activation of the PI3K/Akt/mTORC1 signaling pathway in esophageal squamous cell carcinoma
Authors: Gen Yasuyuki , Yasui Kohichiroh , Kitaichi Tomoko , Tomie Akira , Ito Yoshito
Published: May 2015

Our previous study revealed that SOX2, a master regulator during embryogenesis, is an amplification target of 3q26.3 in esophageal squamous cell carcinoma (ESCC) and that SOX2 promotes ESCC cell proliferation. To identify the mechanisms by which SOX2 promotes proliferation of ESCC cells, we assayed multiple signaling pathways activated by SOX2 using a phosphoprotein array. We determined that SOX2 activated the AKT/mammalian target of rapamycin complex 1 (mTORC1) signaling pathway. Immunoblotting confirmed that SOX2 elevated levels of p-AKT and levels of p-p70S6K and p-4E-BP1 which are direct targets of mTORC1. Effects of SOX2 knockdown, including reduced levels of p-AKT and decreased ESCC cell viability, were reversed with constitutive activation of AKT with PTEN knockdown. SOX2 also promoted in vivo tumor growth of ESCC with AKT/mTORC1 activation in mouse xenografts. LY294002, a PI3K inhibitor, suppressed the ability of SOX2 to enhance tumor growth of ESCC by reducing cell proliferation. Furthermore, tissue microarray analysis showed a positive correlation between expression levels of SOX2 and p-AKT in 61 primary ESCC tumors. Our results suggest that SOX2 promotes ESCC tumor growth via activation of the PI3K/AKT / mTORC1 signaling pathway, which enhances cell proliferation.

Citation Format: Yasuyuki Gen, Kohichiroh Yasui, Tomoko Kitaichi, Akira Tomie, Yoshito Ito. SOX2 promotes tumor growth via activation of the PI3K/Akt/mTORC1 signaling pathway in esophageal squamous cell carcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3525. doi:10.1158/1538-7445.AM2014-3525
Abstract 3531: MicroRNA delivery by bionanoparticles: regulation of the liver cancer susceptibility gene MICA expression in hepatocytes
Authors: Ohno Motoko , Otsuka Motoyuki , Kishikawa Takahiro , Yoshikawa Takeshi , Takata Akemi , Koike Kazuhiko
Published: May 2015

Hepatocellular carcinoma (HCC) is an important issue to public health of the world. We recently reported by genome wide association study (GWAS) analyses that a single nucleotide polymorphism (SNP) in the promoter region of the MHC class I polypeptide-related sequence A (MICA) gene that is significantly associated with the risk of hepatitis-virus-related HCC. Because there was a correlation between the expression levels of MICA in the serum depending on the different alleles at the SNP site and the risk of HCC, regulation of MICA expression levels may be useful in the prevention of HCC. Here we show that MICA protein expression in HCC cells can be regulated by the microRNA (miR) 25-93-106b cluster. By use of luciferase assay and FACS analyses, it was confirmed that forced expression of the miR 25-93-106b cluster significantly suppressed MICA expression, whereas inhibition of this miR cluster increased MICA expression in cells that stably express MICA. Furthermore, to deliver efficiently miRNAs into hepatocytes specifically, we developed bio-nanocapsules containing miR93 inside, utilizing hepatitis B virus envelope L-protein. MICA expression levels were indeed suppressed after the delivery of miR93 by these bionanoparticles into hepatocytes. Besides, the binding ability of NKG2D (a receptor of MICA) and in vivo cell-killing by NK cells were changed proportionally to miRNA-induced MICA expression levels. These results suggest that the modulation of MICA expression levels in hepatocytes by delivering miRNAs using these bionanocapsules may be a novel and effective approach to prevent hepatitis-virus-related HCC.

Citation Format: Motoko Ohno, Motoyuki Otsuka, Takahiro Kishikawa, Takeshi Yoshikawa, Akemi Takata, Kazuhiko Koike. MicroRNA delivery by bionanoparticles: regulation of the liver cancer susceptibility gene MICA expression in hepatocytes. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3531. doi:10.1158/1538-7445.AM2014-3531
Abstract 3535: Epigenetic regulation of miR-31 and miR-205 of apoptosis in prostate cancer
Authors: Zhang Qunshu
Published: May 2015

Prostate cancer is the second leading cause of cancer-related mortality among men in the United States. However, our understanding of the cause of prostate cancer remains elusive. Recent studies have implicated that epigenetics, heritable changes in gene expression caused by mechanisms other than classical genetic alterations, is an important mechanism in tumorigenesis. MicroRNAs function as epigenetic regulators to alter gene expression, and very limited information is available regarding the roles of microRNAs in prostate cancer. Evasion of apoptosis is the hallmark of cancer, and it plays an important role in the growth and development of prostate cancer. We have recently demonstrated that miR-31 and miR-205 are downregulated in advanced prostate cancer. In addition, anti-apoptotic protein E2F6 was identified as the target of miR-31 and contributed resistance to chemotherapy-induced apoptosis. While we found that miR-205 is silenced by gene promoter methylation, the mechanism of the downregulation of miR-31 in prostate cancer remains poorly defined. In this study, we report that EZH2 (Enhancer of zeste 2), a histone methyltransferase and component of the polycomb repressive complex 2 (PRC2), is responsible for miR-31 silencing in prostate cancer. EZH2 plays an essential role in the epigenetic maintenance of the H3K27me3 repressive chromatin mark. Our results have shown that siRNA knockdown of EZH2 and DZNep (3-deazaneplanocin) treatment (an EZH2 inhibitor) in prostate cancer cells increased miR-31 expression. Furthermore, EZH2 is responsible for histone methylation at the promoter of miR-31 gene. In addition, siRNA knockdown of EZH2 and the treatment with DZNep sensitized prostate cancer cells to apoptosis. In contrast, EZH2 overexpression confers resistance to chemotherapy-induced apoptosis in prostate cancer cells. Thus, polycomb protein EZH2 suppresses the expression of miR-31 and consequently contributes to apoptosis resistance in prostate cancer. The results will improve our understanding of the mechanism of drug resistance in prostate cancer cells. Novel strategies targeting EZH2 and miR-31 can be developed to increase drug-induced apoptosis and improve cancer response to chemotherapy.

Citation Format: Qunshu Zhang. Epigenetic regulation of miR-31 and miR-205 of apoptosis in prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3535. doi:10.1158/1538-7445.AM2014-3535
Abstract 3538: Obligate haploinsufficiency of the microRNA-processing molecule dicer1 in a murine colitis-associated tumorigenesis model
Authors: Yoshikawa Takeshi , Otsuka Motoyuki , Koike Kazuhiko
Published: May 2015

Background: A widespread downregulated expression of microRNAs (miRNAs) is commonly observed in human cancers. Similarly, deregulated expression of miRNA-processing pathway components, which results in the reduction of global miRNA expression, may also be associated with tumorigenesis. Thus, we hypothesized that the deregulation of microRNA-processing molecule Dicer1 in intestinal epithelial cells may accelerate intestinal inflammation-associated tumorigenesis.

Methods & Results: We constructed Dicer1 gene-disrupted mice with monoallelic and biallelic loss of its locus. Inflammation-associated colon tumors were induced by azoxymethane (AOM) injection with the treatment by three cycles of Dextran Sulfate Sodium (DSS). 1) Dicer1 protein expression and subsequent mature miRNA levels in the isolated intestinal epithelial cells were inversely correlated with the number of intact Dicer1 alleles. 2) Although the severity of inflammation after the induction in this model was comparable in control and Dicer1-mutant mice, the number of colon tumors was significantly higher in heterozygous mice but not in homozygous mice. 3) Because the expression levels of Dicer1 were retained in tumors and its surrounding tissues even after induction of colitis-associated tumors, the effects of Dicer1 deletion on tumorigenesis were considered as cell-autonomous. 4) While the expression levels of representative oncogenes and tumor suppressor genes were in most cases inversely correlated with the expression levels of Dicer1, some genes were not affected by Dicer1 deletion.

Conclusion: Specific ablation of Dicer1 in intestinal epithelial cells accelerates intestinal inflammation-associated tumorigenesis. However, this effect was apparent only when a single copy of Dicer1 was deleted, but not with complete Dicer1 gene ablation. Generally, complete loss of the function of most tumor suppressor genes is advantageous for tumor development. However, our results suggest that, in case of Dicer1 gene, complete loss is deleterious for cancer development, while its partial inactivation promotes tumor formation. This unique “obligate haploinsufficient role” colitis-associated colon tumor of Dicer and miRNAs needs further attention to elucidate the pathogenesis of tumorigenesis related with deregulated miRNAs.

Citation Format: Takeshi Yoshikawa, Motoyuki Otsuka, Kazuhiko Koike. Obligate haploinsufficiency of the microRNA-processing molecule dicer1 in a murine colitis-associated tumorigenesis model. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3538. doi:10.1158/1538-7445.AM2014-3538
Abstract 3572: Copy number detection using genomics technologies: A comparison between aCGH and NGS
Authors: Vasmatzis George , Feldman Andrew L. , Johnson Sarah H. , Thorland Erik C. , Fonseca Rafael , Braggio Esteban , Gliem Troy J.
Published: May 2015

Introduction: Identification of causal DNA copy number variations (CNVs) is critical to the understanding of cancer development and progression, and is important clinically for accurate prognosis and efficient treatment. T-cell lymphomas are replete with amplifications and deletions that result in over- and under-expression of genes. Array comparative genomic hybridization (aCGH) has been the technology of choice to detect CNVs but recently, it has been proposed that mate-pair Next Generation Sequencing (mpNGS) can produce similar results. Here, we report a comparison between the two techniques.

Methods: We have developed an algorithm based on extreme value theory to identify CNVs across the genome at high resolution from mate-pair NGS data. The algorithm is based on plotting the distribution of read-counts in windows across the genome, and detecting deletions and gains by finding the windows that differ significantly from the distribution of the normal part of the genome. We compared this algorithm with aCGH data to detect CNVs. We ran five cases of T-cell lymphoma with both techniques and compared the results using concordance statistics.

Results: With 50kb lower cutoff for a CNV, mpNGS detected 98% of the cumulative deleted genome and 94% of the amplified/gained genome that were called by aCGH. Most of the discordant CNVs that were detected by only aCGH were in regions of repetitive areas where mapping fails. Conversely, aCGH detected 75% of the cumulative deleted genome and 74% of the amplified/gained genome that were called by mpNGS.

Conclusion: A detailed comparison of these techniques showed that both have pros and cons. In general, mpNGS can detect most CNVs detected by aCGH, as long as they are more than 50KB. Additionally, mpNGS offers confirmatory data, particularly for deletions, based on detection of aberrant mate pairs that map to the rejoined ends of deleted regional. Finally, the mate-pair technology can also detect balanced translocations and can also predict fusion genes.

Citation Format: George Vasmatzis, Andrew L. Feldman, Sarah H. Johnson, Erik C. Thorland, Rafael Fonseca, Esteban Braggio, Troy J. Gliem. Copy number detection using genomics technologies: A comparison between aCGH and NGS. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3572. doi:10.1158/1538-7445.AM2014-3572
Abstract 3577: Reproducible copy number variation patterns among single circulating tumor cells of lung cancer patients
Authors: Ni Xiaohui , Zhuo Minglei , Su Zhe , Duan Jiachun , Gao Yan , Wang Zhijie , Zong Chenghang , Bai Fan , Wang Jie , Xie X. Sunney
Published: May 2015

Circulating tumor cells (CTCs) enter peripheral blood from primary tumors and seed metastases. The genome sequencing of CTCs could offer noninvasive prognosis or even diagnosis, but has been hampered by the low single-cell genome coverage of scarce CTCs. Here, we report the use of the recently developed multiple annealing and looping-based amplification cycles for whole genome amplification of single CTCs from lung cancer patients. We observed characteristic cancer-associated single-nucleotide variations and insertions/deletions in exomes of CTCs. These mutations provided information needed for individualized therapy, such as drug resistance and phenotypic transition, but were heterogeneous from cell to cell. In contrast, every CTC from an individual patient, regardless of the cancer subtypes, exhibited reproducible copy number variation (CNV) patterns, similar to those of the metastatic tumor of the same patient. Interestingly, different patients with the same lung cancer adenocarcinoma (ADC) shared similar CNV patterns in their CTCs. Even more interestingly, patients of small-cell lung cancer have CNV patterns distinctly different from those of ADC patients. Our finding suggests that CNVs at certain genomic loci are selected for the metastasis of cancer. The reproducibility of cancer-specific CNVs offers potential for CTC-based cancer diagnostics.

Citation Format: Xiaohui Ni, Minglei Zhuo, Zhe Su, Jiachun Duan, Yan Gao, Zhijie Wang, Chenghang Zong, Fan Bai, Jie Wang, X. Sunney Xie. Reproducible copy number variation patterns among single circulating tumor cells of lung cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3577. doi:10.1158/1538-7445.AM2014-3577