bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3316: The stem cell marker nestin inhibits growth and invasion of malignant melanoma
Authors: Akiyama Michiko , Matsuda Yoko , Hagio Masahito , Kawahara Kiyoko , Suzuki Taeko , Ishiwata Toshiyuki , Kawana Seiji , Naito Zenya 
Published: August 2012

Abstract
BACKGROUND: Nestin, a class VI intermediate filament protein, was first described as a neural stem cell/progenitor cell marker. Nestin was detected in various neoplasms, and its expression level has relevance to the poor prognosis in some tumors, including malignant melanomas. Recently, nestin was reported as one of the cancer stem cell markers in melanomas. Furthermore, nestin is expressed in circulating tumor cells of melanoma patients, and its expression was found to be correlated with the prognosis. These reports indicate that nestin plays important roles in melanomas, and has the possibility of becoming a therapeutic target in melanomas. In the present study, we used a silencing strategy of nestin to clarify the effectiveness of nestin-targeting therapy in malignant melanomas. METHODS: We constructed a plasmid vector of short hairpin RNA (shRNA) tageting nestin and scramble vector as a negative control. Human malignant melanoma cells, A375 cells, were stably transfected with these vectors. Using nestin-shRNA-transfected cells, scramble vector-transfected cells, and wild-type cells, we analyzed cell growth, sphere formation, and invasion in vitro. Activation of the AKT/MAPK pathway was analyzed using a phosphorylated protein array. To determine the effect of nestin-knockdown in vivo, we performed tail vein injection of transfected and wild-type A375 cells into nude mice, and analyzed the metastatic tumor formation in 5 weeks. RESULTS: Nestin shRNA transfection suppressed cell growth, invasion, and sphere formation of A375 cells in vitro. In the protein array, the suppression of nestin decreased the phosphorylation of AKT and p53. The incidence of liver metastasis in nude mice was 100% in wild-type cell-injected mice, 75% in scramble vector-transfected cell-injected mice, and 16.7% in nestin-shRNA-transfected cell-injected mice. CONCLUSION: These findings suggest that nestin regulates cell growth, stemness, invasion, and metastasis of melanomas, in part, by alterling the phosphorylation of the MAPK/AKT pathway. Nestin may be a candidate therapeutic target in malignant melanomas.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3316. doi:1538-7445.AM2012-3316
Abstract 3319: CD44 standard isoform regulates the mesenchymal phenotype in hepatocellular carcinoma cells, and associates with a poor prognosis in patients.
Authors: Mima Kosuke , Okabe Hirohisa , Ishimoto Takatsugu , Beppu Toru , Baba Hideo 
Published: August 2012

Abstract
Purpose: Cancer stem cells (CSCs) in many tumors have been identified using CD44, either individually or in combination with other markers and are involved in tumor progression and metastasis. Epithelial-mesenchymal transition (EMT) is known to induce cancer invasion, metastasis, and CSCs properties. However, the underlying molecular mechanisms which integrate EMT and CSCs properties remain largely unknown. In this study, we test the hypothesis that CSCs markers play an important role in regulating EMT in hepatocellular carcinoma (HCC). Experimental Design: The relationship between the CSCs markers and EMT markers was investigated in HCC cell lines. The EMT markers, invasive phenotype, the relevant transcriptional factors and signaling pathways induced by CD44 were examined in vitro. The clinicopathological importance of CD44 standard isoform (CD44s) expression was analyzed by an immunohistochemical analysis, and the correlation of CD44s and EMT markers was determined in 150 patients with hepatocellular carcinoma. Results: HCC cell lines with high CD44 expression showed decreased E-cadherin expression and increased vimentin expression. CD44s mRNA was dominant form of CD44 mRNA present in HCC cell lines with the mesenchymal phenotype. Overexpression of CD44s in HCC cells induced the mesenchymal phenotype and increased tumor cell invasion, whereas knockdown of CD44s in HCC cells led to loss the mesenchymal phenotype and decreased tumor cell invasion. CD44s expression was associated with E-cadherin low expression (P=0.039) and vimentin high expression (P&lt;0.001). CD44s expression was related to a large tumor size (P=0.003), multiple tumors (P=0.032), and poor tumor differentiation (P=0.020), and a shorter disease-free (P=0.023) and overall survival (P=0.013) in patients with HCC. Conclusions: our study will be valuable for understanding the relationship between CSCs and EMT, and the identification of new treatment targets for future HCC treatment.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3319. doi:1538-7445.AM2012-3319
Abstract 333: SFRP4 modulates EMT, cell migration and downstream Wnt signalling in epithelial ovarian cancer.
Authors: Ford Caroline E. , Jary Eve , Ma Sean , Nixdorf Sheri , Heinzelmann-Schwarz Viola , Ward Robyn L. 
Published: August 2012

Abstract
Epithelial ovarian cancer is the fifth most common cause of death from all female cancers. This poor overall prognosis results in part from late diagnosis and a current lack of targeted therapy, stemming from the current limited understanding of the initiating events of ovarian carcinogenesis. There is a critical need for the identification of ovarian cancer pathways and targets to aid in diagnosis, as prognostic indicators and as targets for novel therapies. Wnt signalling is essential for crucial components of carcinogenesis and metastasis including differentiation, polarity, migration, adhesion and survival, and hence the role of Wnt signalling in human cancer is increasingly being investigated along with strategies to target pathway components. The Wnt pathway is regulated at multiple levels, with the Wnt antagonists or “gatekeeper proteins” receiving attention in recent years, due to their frequent loss in cancer. Secreted frizzled receptor proteins (SFRPs) are extracellular inhibitors of Wnt signalling that act by binding directly to Wnt ligands or Frizzled receptors, and have been shown to function as tumour suppressors in other cancer types. We have previously shown that one of these inhibitors, SFRP4, is progressively lost in more aggressive ovarian cancer phenotypes, such as Type II cancers (Jacob et al, 2011). Furthermore, patients lacking SFRP4 expression had a poorer prognosis than those expressing SFRP4. Here, we investigated the functional consequences of SFRP4 modulation in ovarian cancer cell lines. Re-expression of SFRP4 resulted in decreased expression of downstream components of Wnt signalling, inhibition of cell migration, and inhibition of epithelial to mesenchymal transition (EMT). In parallel, knockdown of SFRP4 resulted in ovarian cells upregulating markers of EMT. These results indicate that modulation of an upstream gatekeeper of Wnt signalling can have dramatic downstream effects on ovarian cancer cell behaviour, as mediated through EMT. This raises the possibility that SFRP4 may be a suitable marker of ovarian carcinogenesis, and that targeting the Wnt signalling pathway may have potential as an ovarian cancer treatment.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 333. doi:1538-7445.AM2012-333
Abstract 3348: Pax8-PPARδ fusion protein activates β-catenin/TCF pathways
Authors: Vu-Phan Dang L. , Yu Jingcheng , Koenig Ronald J. , Wicha Max S. 
Published: August 2012

Abstract
Pax8-PPARδ Fusion Protein (PPFP) occurs in ∼35% of follicular thyroid carcinomas. Expression of PPFP in the non-transformed rat thyroid cell line PCCL3 conferred on the cells the ability to invade through matrigel and to form colonies in anchorage independent conditions. PPFP also increased the percentage of cells that have activated β-catenin/TCF. We transfected control and PPFP expressing PCCL3 thyroid cell lines with two different β-catenin/TCF-GFP reporter systems. We then sorted a population of GFP positive cells and expanded them to obtain 100% transfected cell lines. These GFP positive cells gave rise to a new heterogenous population with a stable equilibrium between GFP positive, β-catenin/TCF active cells and GFP negative, β-catenin/TCF inactive cells. PPFP expressing cell lines contained more than twice the percentage of GFP positive cells compared to control cell lines. GFP positive PPFP expressing thyroid cells were enriched for cells exhibiting the transformed phenotypes previously observed in the unsorted, untransfected cell lines. Sorted β-catenin/TCF active single cells gave rise to heterogenous clonal populations containing both β-catenin/TCF active and negative cells. Clonal populations derived from β-catenin/TCF inactive cells remained purely negative. Clonal populations derived from GFP positive cells were twice as enriched for anchorage independent colony forming cells than GFP negative clones. More strikingly, GFP positive clones were 5-10 times more invasive than negative clones, which exhibited the same invasive potential as control cells. Treatment with PPARδ full or partial agonists further increased the β-catenin/TCF population in PPFP expressing cells. Again, the effects were confined to populations containing β-catenin/TCF active cells. Co-treatment with PPARδ antagonists reversed the effects of PPARδ agonists. These data suggest that β-catenin/TCF active cells have properties of cancer stem cells and that PPFP acts on this population to expand the stem cell like fraction.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3348. doi:1538-7445.AM2012-3348
Abstract 3350: Implications of the STAT6 pathway by interleukin-4 in prostate cancer
Authors: Erb Holger H. , Santer Frédéric R. , Culig Zoran 
Published: August 2012

Abstract
Cancer-related inflammation (CRI) can promote or inhibit tumor development, growth and progression. A key role in CRI may be the infiltration of immune cells. During the inflammation process immune cells are recruited to the tumor. They produce cytokines and inflammatory molecules like reactive oxygen and nitrogen species. However, in prostate Cancer (PCa) the role of inflammation is not well understood. Meta-analysis conducted on case-control studies showed an association of prostatitis and the risk of prostate cancer. Moreover, clinical data showed that several cytokines e.g. interleukin (IL)-6, IL-8 and IL-4 are significantly elevated in sera of patients with CRPC. In this study we investigated the role of short-term and long-term IL-4 treatment on the castration resistant prostate cancer lines PC3, Du145 and LNCaP-IL6+. For short-term treatment conditions cells were incubated with increasing concentrations of IL-4 (0.1-10 ng/ml) for 96 h in full growth medium. Neither proliferation rate nor cell-viability of the three cell lines were changed under these conditions. On the other hand, from the known IL-4 downstream targets only phosphorylation of STAT6 was readily detectable after IL-4 treatment, whereas phosphorylation status of the MAP kinases ERK1/2 and AKT was not affected. Among the STAT6 regulated genes only SOCS-1 was upregulated after IL-4 treatment. For long-term treatment conditions PC3, Du145 and LNCaP-IL-6+ cell lines were treated with 5 ng/ml IL-4 over a period of 30 passages. Interestingly, a morphologic change of the PC3 cell line to a more dedifferentiated phenotype was observable after 10 passages of IL-4 treatment and peaked after 20 passages with 30-50% of all cells showing this morphology. In order to evaluate possible effects of IL-4 on stem-cell like phenotype, we analyzed the IL-4 treated PC3 cell line for the presence of stem cell markers CD44, CD133 and integrin α2 by FACS. Indeed, high expression of CD44 and integrin α2 was observable in a subpopulation of PC3 cells. Expression of CD133 was negative due to hypermethylation of the gene. The percentage of marker-positive cells was increasing with passage, showing that IL-4 long-term treatment might be favorable for cells with cancer stem cell-like properties. Collectively, our results show that IL-4 long-term treatment might promote cells with stem cell-like characteristics and therefore influence the progression of the cancerous disease. This work was funded by the European Union FP7 Marie Curie Initial Training Network PRO-NEST.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3350. doi:1538-7445.AM2012-3350
Abstract 3353: Nodal signaling mediates the aggressive, tumorigenic phenotype in breast cancer cells
Authors: Kirsammer Gina , Strizzi Luigi , Gilgur Alina , Margaryan Naira , Kirschmann Dawn , Seftor Elisabeth , Hendrix Mary JC 
Published: August 2012

Abstract
The embryonic morphogen Nodal is a potent inducer of EMT and critical mediator of stem cell pluripotency in the developing embryo, but is not expressed in normal adult tissues. Intriguingly, our lab has recently shown that Nodal, a TGF-beta family member, is in fact secreted by melanoma and breast cancer cells and that Nodal expression positively correlates with tumor grade in human patient samples. Our current studies demonstrate a positive role for Nodal in regulating breast cancer cell renewal and aggressiveness. Specifically, short hairpin (shRNA) knockdown of Nodal expression in human breast cancer cell lines diminishes proliferation, invasion and clonogenicity of these cell lines in vitro and dramatically curtails tumor formation in a mouse xenograft model. Additionally, our data suggest that these effects may be caused, in part, by alterations in the expression of adhesion receptors known to function in tumor engraftment and metastasis. Taken together, our data suggest that Nodal signaling may facilitate the acquisition of self-renewal and migratory capabilities in breast cancer cells and represent a novel therapeutic target for disrupting these processes in breast cancer cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3353. doi:1538-7445.AM2012-3353
Abstract 3358: 1α,25-dihydroxyvitamin D3 inhibits the proliferation of cultured glioblastoma multiforme cells
Authors: Pinskey Justine M. , Winn Robert J. 
Published: August 2012

Abstract
Glioblastoma multiforme (GBM) is a common, aggressive type of brain tumor with a median survival of only fifteen months. Brain tumor stem cells (BTSCs) within GBM tumors resist standard treatments, initiate recurrence, and pose a significant challenge for GBM treatment. This study examined Gli1 expression and proliferation of GBM cells in vitro to evaluate the effects of 1α,25-dihydroxyvitamin D3 (vitamin D3). Vitamin D3 is a safe, natural inhibitor of the hedgehog signaling pathway–a mechanism essential to BTSC function. Immunocytochemistry demonstrated that both established GBM cell lines and GBM-derived BTSCs expressed Gli1, indicating hedgehog signaling pathway activity within the cell populations. Vitamin D3 treatment significantly reduced cell proliferation, both in GBM cell lines and GBM-derived BTSCs. Because of active vitamin D3's environmental instability, an in vivo model might provide a better indication of its anti-tumor effects. Ultimately, vitamin D3 may enhance standard GBM treatments by inhibiting hedgehog-expressing cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3358. doi:1538-7445.AM2012-3358
Abstract 336: 4-methylumbeliferone, an HA synthesis inhibitor, inhibits EMT determinants in bladder cancer cell lines
Authors: Yates Travis , Garcia-Roig Mike , Veerapen Muthu K. , Benitez Anaid , Lokeshwar Vinata 
Published: August 2012

Abstract
INTRODUCTION AND OBJECTIVE: Elevated Hyaluronic Acid (HA) levels are a molecular determinant of bladder cancer (BCa) metastasis. Furthermore, HA, HYAL-1 hyaluronidase and HA-synthesizing enzymes are potential markers for BCa diagnosis and predicting prognosis. 4-Methylumbelliferone (4-MU) is an orally bioavailable dietary supplement that inhibits HA synthesis. Epithelial mesenchymal transition (EMT) is the hallmark of invasion and metastasis, in which cancer cells acquire an invasive phenotype. Since HA promotes tumor metastasis, we evaluated the antitumor effects of 4-MU in vitro and in vivo and examined whether 4-MU can reverse the EMT by inhibiting HA synthesis. METHODS: Quantitative PCR was used to measure mRNA expression of EMT genes (β-catenin, Twist, and Snail) in 66 bladder tissue specimens (27 normal; 39 tumor); follow-up: 26±4.3 months; median 20 months. The effect of 4-MU (0.2 - 0.6 mM) on cell proliferation, apoptosis, intracellular signaling, and the expression of HA receptors and EMT genes were examined in BCa cell lines, 253J-Lung and HT1376. Effect of 4-MU on tumor growth was analyzed in subcutaneous xenografts. RESULTS: Among the EMT genes, both Snail and Twist were differentially expressed in BCa tissues when compared to normal bladder (P&lt;0.001). Furthermore, Twist expression significantly correlated with metastasis (+: 0.25+0.15, -: 0.12+0.14; P=0.028). β-catenin expression negatively correlated with survival (chi square=4.3, p=0.038). 4-MU inhibited proliferation and induced apoptosis in the BCa cells. Following a 48 h exposure at the IC50 for HA synthesis (0.4 mM), 4-MU inhibited proliferation by 61% and increased apoptosis by 73%. 4-MU induced caspase-8, -9 and -3 activation and up-regulation of Fas and FADD. 4-MU downregulated the expression of HA-receptors - CD44, RHAMM, EMT promoters - β-catenin, Snail and Twist by 4-10-fold, but increased the expression of E-cadherin by 2-fold, suggesting EMT-reversal. In xenograft studies, 4-MU significantly decreased tumor growth (&gt; 3-fold) when treatment was started either on the day of tumor cell injection or after tumors became palpable. No weight loss or serum or organ toxicity was observed in treated mice. Tumors showed reduced microvessel density (∼3-fold) and HA expression but increased TURNEL positive cells. CONCLUSION: This is the first study that shows inhibition of HA synthesis by an orally bioavailable, non-toxic supplement potentially reverses EMT and has potent anti-tumor activity. Support: R01 CA 123063-04 (VBL); R01 CA 72821-11 (VBL).
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 336. doi:1538-7445.AM2012-336
Abstract 3381: Photochemical internalization (PCI) of immunotoxins targeting cancer stem cell markers
Authors: Bostad Monica , Berg Kristian , Høgset Anders , Selbo Pål Kristian 
Published: August 2012

Abstract
The cancer stem cell (CSC) theory indicates that only a subset of the cancer cells is capable of tumor-initiation, self renewal and differentiation. CSC is also thought to be treatment resistant and responsible for recurrence. Different antigens are being used to identify CSC markers and are potential targets for CSC-based therapy. Due to the expression of the same antigens on normal stem cells it is of paramount importance that CSC-targeting cytotoxins are specifically and efficiently delivered to malignant tissues. Photochemical internalization (PCI) is a method for specific delivery of membrane impermeable macromolecules from endocytic vesicles to the cytosol of targeted cells. In this study we show that PCI increase the cytotoxic effect of immunotoxins (IT) targeting stem cell markers on different cell lines. The IT consists of a CSC-targeting mAb biotinylated to the ribosome inactivating plant (RIP) toxin saporin. TPCS2a-PCI of one of the ITs showed to be very cytotoxic at sub picomolar levels. Cytotoxicity of the IT was blocked by using excess concentration of unconjugated Ab. In addition, no difference in cytotoxicity was obtained between mAb-toxin and toxin alone in marker negative cell lines, confirming specificity. In order to asses enrichment of CSC-activity in vitro we used flow cytometry to sort cells based on the degree of expression of CSC-markers and compared the ability of different cell fractions to establish colonies in both 2D and 3D cell cultures growing in serum-free stem cell media. Cells with high expression of CSC-markers had significantly higher capacity to initiate colonies than low expressing cells. CSC enrichment was confirmed by an in vivo limiting dilution assay. The present study demonstrates that PCI is an efficient method for selective killing of cancer cells expressing stem cell markers and will be further explored for the targeting of tumors that express these markers.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3381. doi:1538-7445.AM2012-3381
Abstract 3849: Triptolide reduces activity of RNA polymerase II
Authors: Yang Shanmin , Huang Luqiang , Chen Chun , Zhang Mei , Zhang Zhenhuan , Han Deping , Hong Jinsheng , Yin Liangjie , Zhang Bingrong , Zhang Amy , Vidyasagar Sadasivan , Okunieff Paul , Zhang Lurong 
Published: August 2012

Abstract
Triptolide (TPL) possesses potent anticancer and anti-inflammatory effects. Our previous study demonstrated that radiation-induced pulmonary fibrosis in mice can be significantly reduced by TPL treatment (0.25 mg/kg/every 3 days for 1-3 months via intravenous injection). Treatment was associated with the reduction of a panel of inflammation molecules, as evidenced by a membrane array of 62 inflammatory molecules. We found that TPL has a high level of biointeraction with RNA polymerase II (Pol II). Pol II is a major enzyme complex responsible for the production of mRNA and could explain improved tumor response and reduced normal tissue toxicity. The mRNA of interleukin-1 beta (IL-1α), tumor necrosis factor-alpha (TNFα), IL-6, and Cox-2 were used as downstream products of Pol II. Alpha-amanitin (a specific inhibitor of Pol II) and actinomycin D (a pan Pol inhibitor) were used as positive controls. The results showed that: 1) Pol II was suppressed as early as 1 hour after TPL was added to cells; 2) TPL (effective at ng/ml levels) is more potent than α-amanitin and actinomycin D (effective at µg/ml levels) at reducing housekeeping genes L32 and S14; 3) the suppression of Pol II by TPL was dose dependent; and 4) TPL inhibited radiation-induced mRNA of IL-1α and TNFα at 3 hours after irradiation and IL-6 and Cox-2 at 8 hours after irradiation. It is unknown whether these early effects of TPL on Pol II impact either tumor response or normal tissue complications.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3849. doi:1538-7445.AM2012-3849