bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4018: Development and analytical validation of a novel real-time multiplex RT-qPCR assay for the simultaneous quantification of ER, PR, HER-2 and EGFR mRNA expression in circulating tumor cells of breast cancer patients
Authors: Strati Areti D. , Lianidou Evi S. 
Published: May 2015

Abstract
Introduction: Circulating Tumor Cells (CTCs) represent an important biological link in the spread of breast cancer from primary to metastatic disease. CTCs have already been established as strong predictors of prognosis in patients with metastatic breast cancer. Triple-negative breast cancers have a more aggressive clinical outcome than other forms of breast cancer. The aim of our work was to develop a quantitative real-time multiplex RT-qPCR assay for studying the expression of ER, PR, HER-2 and EGFR in CTC of breast cancer patients.
Materials and Methods: A quadruplex quantitative real time RT-qPCR assay for ER, PR, HER-2 and EGFR was developed based on the de novo in-silico design of eight primers and four dual hybridization probes. All primers and probes were designed to avoid cross-reactions and primer dimer formation and to match the assay conditions, such as amplicon sizes, and melting temperatures. The specificity of all primer and hybridization probe sequences was first tested by homology searches in the nucleotide database (NCBI, nucleotide BLAST). Each probe set included a 3′-fluorescein (F) donor probe and a 5′- LC acceptor probe that was different for each gene set: ER at 610nm, PR at 640nm, HER-2 at 670nm, EGFR at 705nm. A color compensation test was performed by using pure dye spectra so that spectral overlap between dyes was corrected. The LightCycler 2.0 platform, (Roche, Diagnostics) was used since it allows detection of six multiple reporter dyes in the same capillary. The assay was designed, so that only 1 μL of cDNA is used and the total reaction volume is 10 μL.
Results: The assay sensitivity was evaluated by performing spiking experiments. For this reason known numbers of MCF7 and SKBR-3 cells, as well as external quantification calibrators ranging from 105 copies/μL to 10 copies/μL, were used. The specificity was evaluated by carefully designed cross reaction studies, performed for each gene target in the presence of all other targets. Our assay, under optimized conditions has shown a very high specificity for each analyte.
Conclusions: We report for the first time a highly sensitive, specific and reproducible quantitative multiplex real-time RT-qPCR assay for the simultaneous quantification of ER, PR, HER-2 and EGFR. The assay is currently validated in CTC isolated from a large number of breast cancer patients.
Citation Format: Areti D. Strati, Evi S. Lianidou. Development and analytical validation of a novel real-time multiplex RT-qPCR assay for the simultaneous quantification of ER, PR, HER-2 and EGFR mRNA expression in circulating tumor cells of breast 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 4018. doi:10.1158/1538-7445.AM2014-4018
Abstract 4020: Exploring target gene(s) within chromosome 19-amplification detected in a subclone from metastatic tumors in mouse transplantable OSCC
Authors: Muramatsu Tomoki , Kozaki Ken-ichi , Imoto Seiya , Yamaguchi Rui , Tsuda Hitoshi , Kawano Tatsuyuki , Miyano Satoru , Inazawa Johji 
Published: May 2015

Abstract
Cancer metastasis is caused by a highly complex molecular and cellular mechanism. Although this mechanism has been gradually revealed, there are few genes found as useful diagnostic markers or molecular targets for clinical application so far. In this study, we attempted to establish highly metastatic cells by in vivo selection for identifying metastasis-related genes with an oral squamous cell carcinoma cell line (HOC313-parent). As a result, we established a highly metastatic subline (HOC313-LM) derived from a metastatic site of axillary lymph node. To explore metastasis-related genes and signaling pathways, we performed integrative omics including several genomic and epigenomic approaches combined with systems biology. Since the amplification of chromosome 19 was identified in HOC313-LM by array CGH, we focused on this amplicon containing 44 genes. Among them, one gene, which was essential for the posttranslational modification, was associated with tumor formation and metastasis in vivo via the Rho signaling pathway. Indeed, activation of the Rho signaling pathway emerged in HOC313-LM by our systems biology approach using the gene expression data sets. In addition, the expression pattern of this gene correlated with poor disease-free survival and distant metastasis in patients with esophageal squamous cell carcinoma. Then, we performed therapeutic experiments using a known inhibitor of this gene, and found that tumor growth was inhibited by this compound. Taken together, our findings suggested that this gene might serve as a novel molecular target for anti-cancer therapy.
Citation Format: Tomoki Muramatsu, Ken-ichi Kozaki, Seiya Imoto, Rui Yamaguchi, Hitoshi Tsuda, Tatsuyuki Kawano, Satoru Miyano, Johji Inazawa. Exploring target gene(s) within chromosome 19-amplification detected in a subclone from metastatic tumors in mouse transplantable OSCC. [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 4020. doi:10.1158/1538-7445.AM2014-4020
Abstract 4026: Attenuation of pancreatic cancer cell migration and invasion through a targeted inhibition of the Rac GEF Vav1
Authors: Razidlo Gina L. , Magnine Christopher , Sletten Arthur C. , Hurley Rachel M. , McNiven Mark A. 
Published: May 2015

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal forms of human cancer, due in large part to a high incidence of metastasis and resistance to conventional chemotherapies. As such, new therapies are needed to halt the metastatic process. The proto-oncogene Vav1 is ectopically expressed in over half of human pancreatic cancers, and its expression correlates with a poor prognosis in humans and promotes survival and transformation in isolated tumor cells. We have found that Vav1, through its GEF activity towards Rac1, also potently induces migration by pancreatic tumor cells. In addition, Vav1 also signals through Cdc42 to drive the formation of invadopodia and matrix degradation, which also promotes tumor cell invasion. Importantly, the activation of Vav1 circumvents the requirement for active Src in this process. Because Vav1 is ectopically expressed in pancreatic cancers and is a potent regulator of tumor cell survival, proliferation, and invasive migration, it could provide a specific therapeutic target for these tumors. Recently, the anti-inflammatory agent azathioprine was described as an inhibitor of Vav1/Rac1 activity in hematopoietic cells. Therefore, we hypothesized that azathioprine could also inhibit the Vav1-driven processes of proliferation and invasion in pancreatic tumor cells. Indeed, we have determined that azathioprine treatment of cultured human PDAC cells reduces viability and proliferation. Further, similar to siRNA-mediated depletion of Vav1, azathioprine treatment reduces cell migration, as well as the matrix degradation required for invasion. Vav1-expressing tumor cells were more sensitive to azathioprine than Vav1-negative cells, supporting the premise that the inhibitory effects of azathioprine are mediated through Vav1. Finally, azathioprine also reduces metastasis in a xenograft model of pancreatic cancer. Taken together, these data uncover a role for aberrantly expressed Vav1 in regulation of the cytoskeletal machinery required for migration and invasion, and in the promotion and metastasis of pancreatic cancer. Even with the expression of multiple Rho family GEFs, tumor cells that upregulate Vav1 have become dependent upon this exchange factor, implicating Vav1 as a potent node for therapeutic intervention. Supported by R01 CA104125 and R03 CA155778 to M.A.M.
Citation Format: Gina L. Razidlo, Christopher Magnine, Arthur C. Sletten, Rachel M. Hurley, Mark A. McNiven. Attenuation of pancreatic cancer cell migration and invasion through a targeted inhibition of the Rac GEF Vav1. [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 4026. doi:10.1158/1538-7445.AM2014-4026
Abstract 4034: The anti-peritoneal metastasis properties of Yangzheng Xiaoji, the potential role of hyaluronan and CD44
Authors: Jiang Wen G. , Jia Yongning , Ye Lin , Ji Ke , Gao Yong , Wei Cong , Wu Yiling , Topley Nick , Kift-Morgan Anna , Ji Jiafu 
Published: May 2015

Abstract
Introduction. Peritoneal metastasis is a frequently occurring complication patients with gastrointestinal, pancreatic, ovarian cancers and is a condition difficult to manage, clinically, due largely to the lack of effective treatment. In the present study, we report an accidental and novel discovery that the extract of Yangzheng Xiaoji (YZXJ), a traditional medical formula used in cancer treatment had an anti-peritoneal metastasis function.
Materials and methods. Human primary cultured peritoneal mesothelial cells were used as an in vitro cellular model. Human gastric, colorectal, pancreatic, ovarian and prostate cancer cells were used in the tests. Tumour-mesothelial interaction, invasion and migration were assessed with YZXJ. Athymic nude mice peritoneal metastasis model was used to evaluate the effects in vivo.
Results. Soluble YZXJ was firstly extracted in a highly reproducible manner. YZXJ extract (DME25) had a negletable toxicity to cancer cells and mesothelial cells. However, it profoundly inhibited the tumour-mesothelial adhesions in the type of tumour cells tested with pancreatic and colorectal cancer cells amongst the most sensitive ones to YZXJ (p&lt;0.001), a phenomenon seen with a number of cell based assays. The inhibitory effect was also seen with cellular migration. A similar pattern of inhibition was seen on tumour adhesion to hyaluronan (HA) coated surface. It was further revealed that YZXJ and an hyaluronan inhibitor and anti-CD44 antibodies had additive effects with YZXJ on tumour-mesothelial interactions. YZXJ extract had an inhibitory effect on the phosphorylation of SRC kinase in cancer cells and in mesothelial cells as shown by Western blotting and immunofluorescence microscopy. In the peritoneal metastasis in vivo model, it was demonstrated that intraperitoneal administration of YZXJ significantly reduced the formation of metastatic nodules (p=0.035, control vs YZXJ treatment). This was seen with little side effects. We have also shown that hyaluronan inhibitor strengthened the inhibitory effect exerted by YZXJ.
Conclusion. Yangzheng Xiaoji has an potent inhibitory effect on tumour-mesothelial interactions of a variety of tumour types and also markedly reduces the peritoneal metastasis in vivo. This effect is likely to be achieved by its effect on the SRC signalling pathway. Yangzheng Xiaoji presents a new way of treating peritoneal metastasis.
Citation Format: Wen G. Jiang, Yongning Jia, Lin Ye, Ke Ji, Yong Gao, Cong Wei, Yiling Wu, Nick Topley, Anna Kift-Morgan, Jiafu Ji. The anti-peritoneal metastasis properties of Yangzheng Xiaoji, the potential role of hyaluronan and CD44. [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 4034. doi:10.1158/1538-7445.AM2014-4034
Abstract 4037: Combining tyrosine kinase inhibitors and isoflavones to target metastatic activity in prostate cancer cells
Authors: Rice Lori P. , Pampo Christine , Lepler Sharon , Siemann Dietmar W. 
Published: May 2015

Abstract
Background: Efforts to impede early steps in the metastatic cascade have led to the development of promising small molecule targeting agents. The purpose of this study was to evaluate the use of Src tyrosine kinase inhibitors known to interfere with cell migration and invasion, in combination with a cytostatic soy isoflavone extract (ISFs) on metastatic activity in prostate cancer cells. Methods: PC-3ML cells (a gift from A. Fatatis, Drexel University), and LNCaP cells treated with a Src inhibitor, ISFs, or a combination, were evaluated for clonogenic cell survival, cell cycle progression, and transwell migration and invasion activity. Results: Src inhibitors and ISFs induced an accumulation of cells in the G1 and G2/M phases of the cell cycle, respectively, with little change in cell viability or survival. Invasion studies with PC-3 ML and LNCaP cells suggest that ISFs can reduce metastatic activity by 39% and 35%, respectively. In addition, ISFs enhance the effectiveness of the Src inhibitors to reduce invasiveness. This may indicate that including ISFs in treatment regimens may allow a lower dose of the targeting agent to be used to achieve optimal response and decrease toxicity. Conclusions: In vitro studies suggested that combining a Src inhibitor and ISFs resulted in greater inhibition of metastatic activity than either alone.
Citation Format: Lori P. Rice, Christine Pampo, Sharon Lepler, Dietmar W. Siemann. Combining tyrosine kinase inhibitors and isoflavones to target metastatic activity in 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 4037. doi:10.1158/1538-7445.AM2014-4037
Abstract 404: Function of Brg1 chromatin remodeling factor in sonic hedgehog-dependent medulloblastoma development
Authors: Shi Xuanming , Zhang Zilai , Wang Qiu , Wu Jiang 
Published: May 2015

Abstract
Medulloblastoma is the most common malignant pediatric brain tumor. Overactive Shh signaling in cerebellum granule neuron precursors (CGNPs) is the leading cause of the childhood medulloblastoma (Shh-subtype). Previously we have shown that chromatin remodeler Brg1 is required for Shh target gene expression, and Brg1 deletion reduced CGNP proliferation in developing cerebellum. Current study focuses on the function of Brg1 in mouse model of Shh-subtype medulloblastoma. In CGNP cultured from SmoM2 transgenic mice where Shh pathway is constitutively active, we found Brg1 is required for CGNP mitogenic target gene expression and proliferation. In SmoM2 medulloblastoma cultures, we observed that tumor cell growth was inhibited by conditional knockout of Brg1. In subcutaneous transplantation, we found that tumors were significantly shrunk upon induction of Brg1 deletion. Detailed analysis indicated that Shh-dependent mitogenic target genes decreased by loss of Brg1. Further evidences showed that medulloblastoma cell proliferation was significant inhibited by conditional knockout of Brg1. Effect of Brg1 on the chromatin environment of target genes during medulloblastoma development will be discussed. Our study will provide insights to the epigenetic mechanism of Shh-dependent tumor development and new therapeutic targets.
Citation Format: Xuanming Shi, Zilai Zhang, Qiu Wang, Jiang Wu. Function of Brg1 chromatin remodeling factor in sonic hedgehog-dependent medulloblastoma development. [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 404. doi:10.1158/1538-7445.AM2014-404
Abstract 4044: Effect of adjuvant chemotherapy on the phenotype of circulating tumor cells in patients with triple negative breast cancer
Authors: Kallergi Galatea , Dragolia Melina , Mavroudis Dimitris , Georgoulias Vassilis , Agelaki Sofia 
Published: May 2015

Abstract
Introduction: Recent studies revealed phenotypic discrepancies between Circulating Tumor Cells' phenotype (CTCs) and the corresponding primary tumor in patients with breast cancer. Patients with triple negative (TN) (ER-negative, PR-negative, HER2-negative) primary tumors are considered as a poor prognosis group for whom no targeted therapies are currently available. The aim of the current study was to characterize CTCs in TN breast cancer (BC) patients before and after adjuvant chemotherapy in order to explore additional therapeutic approaches based on the phenotype of chemoresistant CTCs.
Methods: Cytospins were prepared from peripheral blood mononuclear cells (PBMC), obtained by ficoll density gradient centrifugation of 31 TN BC patients, before and after adjuvant treatment. The expression of Cytokeratins (CK), Estrogen Receptor (ER), Progesterone Receptor (PR), EGFR and HER2 in CTCs was assessed using double immunofluorescent experiments and ARIOL analysis.
Results: CTCs were detected in 25 out of 31 (81%) patients before and in 18 out of 31 (58%) after adjuvant chemotherapy. ER, PR, EGFR and HER2 expression was detected in 33.7%, 23.0%, 69.3%, 42.8% of the total examined CTCs before adjuvant chemotherapy and in 4.2%, 0%, 61.79% and 15.73% after treatment. The frequency of ER- and PR- expressing CTCs was significantly (p=0.05, p=0.007) reduced post-chemotherapy. The absolute number of ER and PR positive-CTCs per patient was also statistically decreased (p=0.02, p=0.001) Triple staining experiments could not reveal co-expression of CK/EGFR/ER in CTCs; however, there was one patient with CTCs co-expressing CK/HER2/ER.
Conclusions: A significant percentage of CTCs in triple negative breast cancer patients express HER2 and EGFR before treatment, implying that these receptors could be a potential target for the limitation of metastasis in these patients. EGFR-expressing CTCs persist after adjuvant chemotherapy, suggesting that additional treatment with EGFR-targeting agents could be used post-chemotherapy to eliminate chemo-resistant CTCs.
Citation Format: Galatea Kallergi, Melina Dragolia, Dimitris Mavroudis, Vassilis Georgoulias, Sofia Agelaki. Effect of adjuvant chemotherapy on the phenotype of circulating tumor cells in patients with triple negative 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 4044. doi:10.1158/1538-7445.AM2014-4044
Abstract 4047: Potential role of CXCR6-CXCL16 in prostate cancer progression and chemotherapeutic efficacy
Authors: Gupta Pranav , Ward Ashley B. , Mir Hina , Kaur Gurpreet , Grizzle William E. , Lillard James W. , Singh Shailesh 
Published: May 2015

Abstract
Despite recent advances in treatment and clinical management, prostate cancer is still a leading cause of cancer related deaths among men, primarily because etiopathogenesis of PCa is not well defined. In this regard, chemokines and their corresponding receptors have been shown to play major role in PCa progression, which are often associated with poor therapeutic outcomes. Hence, the focus of this study is to determine the potential role of CXCR6 and its natural ligand CXCL16 in PCa pathogenesis, and potential impact of CXCR6-CXCL16 axis on efficacy of docetaxel, which is currently offered in the clinics as standard care.
We investigated expression of CXCR6 and CXCL16 in clinical samples, prostate cancer cell lines and normal prostatic epithelial cells. Expression of CXCR6 and CXCL16 was significantly higher in PCa samples compared to their respective controls, and expression of CXCR6/ CXCL16 was correlated with tumor stage and grades. Similar to patient's samples, expression of CXCR6/CXCL16 was significantly higher in PCa cell lines compared to normal prostatic epithelial cells. Furthermore, levels of phospho -ERK1/2 and -NF-kB, known to be involved in cell growth and survival, were significantly higher in PCa cells treated with CXCL16 compared to untreated controls. In addition to these, role of CXCR6- CXCL16 on genes responsible for cellular adhesion and epithelial-mesenchymal transition were examined. Interestingly, expression of surface E-cadherin and β−catenin following CXCL16 treatment was significantly inhibited compared to untreated cells, suggesting the involvement of CXCR6-CXCL16 interaction in PCa cell migration. Additionally, CXCR6 expressing PCa cells expressed higher α−smooth muscle actin protein, another EMT marker presumably induced by elevated TGF-β, following CXCL16 stimulation. Furthermore, expression of CXCR6, CXCL16 and MMP(s) were elevated in PCa cell following docetaxel treatment in a time and dose dependent manner compared to controls. This may explain the increased efficacy of docetaxel when used in combination with anti-CXCR6 antibody as compared to docetaxel alone. These findings suggest that CXCR6-CXCL16 axis plays a crucial role in pathogenesis of PCa, and inhibition of this axis may have significant impact on disease progression and therapeutics outcomes.
Citation Format: Pranav Gupta, Ashley B. Ward, Hina Mir, Gurpreet Kaur, William E. Grizzle, James W. Lillard, Shailesh Singh. Potential role of CXCR6-CXCL16 in prostate cancer progression and chemotherapeutic efficacy. [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 4047. doi:10.1158/1538-7445.AM2014-4047
Abstract 4055: The transcriptional regulator TBX3 promotes progression of cells representing early premalignant breast cancer
Authors: MacMillan Connor D. , Leong Hon S. , Clifford Allen G. , Krstic Milica , Pardhan Siddika , Dales David W. , Postenka Carl O. , Chambers Ann F. , Tuck Alan B. 
Published: May 2015

Abstract
The transcriptional regulator TBX3 has been shown to be elevated in metastatic breast cancer and is thought to be involved in promoting malignancy of breast and other carcinomas. In a series of cell lines derived from the same patient with breast cancer, we previously showed that metastatic 21MT-1 cells express elevated levels of TBX3, while low levels are expressed by the ductal carcinoma in situ-like 21NT cells. Here we hypothesized that upregulation of TBX3 in 21NT cells would promote their progression to a more malignant phenotype. We found that 21NT cells engineered to overexpress either isoform of TBX3 (iso1, iso2) have an altered growth in 3D Matrigel, with increased colony-forming ability, higher number of cells per colony and less spherical, more irregular colonies, consistent with increased invasiveness. Transwell assays through Matrigel showed increased invasiveness of 21NT cells overexpressing either TBX3 isoform and in vivo assay in the chick embryo similarly showed increased propensity of TBX3 over-expressing 21NT cells for extravasation. These TBX3-induced changes were associated with increased vimentin expression, but minimal change in e-cadherin expression. Overall, these results indicate that TBX3 (iso1 or iso2) expression can promote progression in a model of early breast cancer, by altering cell properties involved in cell survival/colony formation and invasiveness.
Citation Format: Connor D. MacMillan, Hon S. Leong, Allen G. Clifford, Milica Krstic, Siddika Pardhan, David W. Dales, Carl O. Postenka, Ann F. Chambers, Alan B. Tuck. The transcriptional regulator TBX3 promotes progression of cells representing early premalignant 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 4055. doi:10.1158/1538-7445.AM2014-4055