bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3791: Sulphoraphane inhibits STAT3 and its related genes in Du 145 cancer cells
Authors: Koh Wonil , Ahn Kwangseok , Jeong Soojin , Kwon Taerin , kim Jihyun , Shin Jimin , Yun Sunmi , Jung Deokbeom , Jung Jihoon , Lee Eunok , Lee Hyojeong , Lee Hyojung , Lu Junxuan , Kim Sung Hoon 
Published: February 2011

Abstract
Sulforaphane is a naturally occurring isothiocyanate compound mainly found in cruciferous vegetables. Sulforaphane has demonstrated cancer chemopreventive activities in a number of organ sites, including prostate. Mechanistic investigations have shown that SFN induces caspase-mediated apoptosis that is mediated through a rapid intracellular ROS generation. Since STAT3 signaling is crucial for cancer cell survival and ROS has been linked to inactivation of this transcriptional factor, we hypothesized that STAT3 may be an important target for SFN. Here, we investigated the effect of sulforaphane on the regulation of STAT3 in association of ROS in Du145 prostate cancer cells. We first confirmed that sulforaphane induced apoptosis by activation of caspase 8, 9, and 3, PARP cleavage and increase of TUNEL positive cells in Du145 cells, which were preceded for many hours by the increase of intracellular ROS, peaking at 30 minutes of exposure. We demonstrate for the first time that sulforaphane suppressed constitutive STAT3 activation and DNA-binding of STAT3 in a time- and dose-dependent manner in Du145 cells. These actions were also found in U266 multiple myeloma cells and U937 lymphoma cells. In addition, sulforaphane down-regulated the expression of STAT3-related genes such as bcl-2, bcl-xL, survivin, cyclin D1, and VEGF in Du145 cells. Furthermore, sulforaphane cross-linked JAK2, the upstream kinase of STAT3, in a time- and dose-dependent manner. Thiol-reducing agent dithiothreitol (DTT) reversed the inactivation of STAT3 and JAK2 induced by sulforaphane. Interestingly, N-acetyl cysteine (NAC), which leads to increased intracellular GSH synthesis, reversed the molecular patterns of PARP cleavage and inactivation of JAK2 and STAT3 induced by sulphoraphane. Taken together, these findings suggest that sulforaphane exposure leads to a rapid generation of ROS in Du145 cells and inhibition of JAK2/STAT3 pathway, followed by caspase-mediated apoptosis. Our data suggest that the JAK2/STAT3 pathway may be a novel target of SFN contributing to apoptosis.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3791.
Abstract 3795: STAT-3: A Novel Target of Diindolylmethane in Ovarian Cancer
Authors: Kandala Prabodh K. , Srivastava Sanjay K. 
Published: February 2011

Abstract
Signal transducers and activation of transcription (STATs) are the important for growth and development. However defects in STAT's may lead to malignant transformation. STAT-3 is an oncogene which is over expressed in several cancers including ovarian cancer. We have previously demonstrated that Diindolylmethane (DIM), an active metabolite of Indole-3-Carbinol present in cruciferous vegetables, is effective in suppressing the proliferation of human ovarian cancer cells, however, the mechanism through which DIM reduces the growth of cancer cells was not clear. We hypothesized that DIM inhibits the growth of ovarian cancer cells by inhibiting STAT-3 activation. Our present studies demonstrate that DIM treatment inhibit the phosphorylation of STAT-3 at Tyr-705 and Ser-727 in a concentration-dependent manner in SKOV-3 and OVCAR-3 cells. Interestingly, DIM treatment also reduced the constitutive protein expression of STAT-3 in both the cell lines. In a time dependent study, we observed that STAT-3 inhibition was as early as 1h after DIM treatment. Next, we wanted to see whether DIM can block IL-6 mediated activation of STAT-3. Treatment of SKOV-3 or OVCAR-3 cells with 5ng/ml IL-6 for 15min increase the activation of STAT-3 by phosphorylation at Tyr705 by almost twenty fold. Our results reveal that DIM treatment almost completely blocked IL-6 induced phosphorylation of STAT-3 at Tyr705 in both SKOV-3 and OVCAR-3 cells and completely in TOV-21G cells. STAT-3 after activation translocate to nucleus, bind to DNA and is involved in the transcription of responsive genes such as Mcl-1, survivin and Bcl-2. Our results demonstrate that DIM inhibits the DNA binding activity of STAT-3 resulting in the reduced expression of Mcl-1, survivin and Bcl-2. Overexpression of STAT-3 by gene transfection abrogates the cell growth inhibitory effects of DIM. Taken together, our cellular results establish STAT-3 as a therapeutic target of DIM in ovarian cancer. Furthermore, our just completed in vivo studies indicate that oral administration of 2mg DIM per day for 6 weeks significantly suppressed the growth of SKOV-3 tumor xenografts in athymic nude mice. Whether SKOV-3 ovarian tumor growth suppression is through inhibition of STAT-3 in vivo is currently under investigation. [Supported in part by RO1 CA 106953 and CA 129038 awarded to S.K.S by NIH]
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3795.
Abstract 3807: Gugulipid-induced apoptosis in human prostate cancer cells is regulated by Bax/Bak and Bcl-xL
Authors: Zeng Yan , Badmaev Vladmir , Singh Shivendra V. , Xiao Dong 
Published: February 2011

Abstract
Indian Ayurvedic medicinal plant Commiphora mukul has been safely used for treatment of different disorders for thousands of years. Our previous studies have shown that the extract of Commiphora mukul, gugulipid (GL), inhibits growth of human prostate cancer (Pca) cells, but not a normal prostate epithelial cell line (PrEC), by causing apoptosis. We also showed that GL-mediated inhibition of Pca cell proliferation correlated with reactive oxygen species (ROS) production. The present study was designed to gain insight into the mechanism of GL-induced apoptosis downstream of ROS production. We found that the GL-induced apoptosis correlated with induction of proapoptotic proteins Bax and Bak and down-regulation of antiapoptotic proteins Bcl-2 and Bcl-xL in LNCaP and PC-3 cells. The SV40-immortalized mouse embryonic fibroblasts derived from Bax and Bak double knockout mice were significantly more resistant to GL-induced apoptotic cell death (DNA fragmentation and cleavage of PARP) compared with wild-type mouse embryonic fibroblasts. Bcl-xL overexpression in PC-3 cells conferred protection against GL-induced apoptotic cell death. The GL treatment caused activation of c-Jun-N-terminal kinase (JNK), which functions upstream of Bax activation in apoptotic response to various stimuli. Pharmacological inhibition of JNK conferred partial yet significant protection against GL-induced apoptosis. The GL-mediated apoptosis and conformational change of Bax was markedly suppressed by ectopic expression of catalytically inactive mutant of JNK kinase JNKK2 (AA) in LNCaP cells. In conclusion, the present study indicates that Bax/Bak and Bcl-xL regulate GL-induced apoptosis in human prostate cancer cells. This investigation was supported by USPHS grant R21 CA143104-01.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3807.
Abstract 3809: Tumor-released GPI-anchored proteins bind to mannose receptor and polarize tumor-infiltrating macrophages
Authors: Dangaj Denarda , Zhao Aizhi , Nunez-Cruz Selene , Sandaltzopoulos Raphael , Scholler Nathalie 
Published: February 2011

Abstract
Tumors polarize their microenvironment to escape immune-mediated destruction and to promote their growth. Macrophages respond to microenvironmental signals and can be activated toward a classical (M1) or an alternative (M2) phenotype. Tumor-associated macrophages (TAM) resemble to M2 macrophages with a phenotype characterized by the overexpression of IL10, scavenger receptor SR-A, CD68 and MR phenotype but hampered levels of IL-12. M2, in contrast to M1 macrophages, promote TH2 immune responses as well as tissue remodelling and M2 infiltration in solid tumors has been shown to significantly correlate with poor prognosis. Recent evidences suggest that macrophage differentiation towards TAM involves a “chemical conversation” via exchange of unknown soluble extracellular mediators between tumor cells and macrophages. In addition, previous work showed that the cross-linking mannose receptor (MR) with an anti-MR mAb could cause dendritic cell differentiation into APCs promoting T-cell anergy.
We hypothesized that MR expressed by macrophages could be bound by tumor-released GPI-anchored glycoproteins through the GPI-anchor mannose residues freed by cleavage. We also hypothesized that MR binding by GPI-anchor could trigger macrophage polarization toward TAMs. To test our hypotheses, we established an in vitro model system of cell co-culture in transwell allowing chemical exchanges between human monocyte-derived macrophages and human ovarian cancer cell line Ovcar3 that sheds mesothelin, a cancer biomarker and a GPI-anchored glycoprotein. Macrophage phenotype changes were monitored by flow cytometry and qRT-PCR. We also isolated three novel recombinant antibodies (scFv) against MR by magnetic and flow sortings from a novel yeast-display human scFv library. We describe here for the first time that tumor-released mesothelin bound to macrophage cell surface in presence of calcium, and that mesothelin binding to macrophages could be blocked by the anti-MR scFv. Furthermore, macrophage phenotype switch toward M2 during coculture with tumor cells could be prevented by one of the anti-MR scFv. Finally, we demonstrated that M1 phenotype could be rescued during coculture with tumor cells with one of the anti-MR scFv. To further characterize the role of MR in vivo, we have implanted mannose receptor knock-out mice intraperitoneally (IP) with luminescent-labeled ovarian cancer cells that overexpress mesothelin. We are now monitoring the tumor growth by in vivo imaging.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3809.
Abstract 3813: Recruitment of macrophages, NK cells, and dendritic cells with metronomic cyclophosphamide-induced regression of gliosarcoma xenografts
Authors: Doloff Joshua C. , Waxman David J. 
Published: February 2011

Abstract
Metronomic chemotherapy displays improved therapeutic activity over conventional schedules for several malignancies in both preclinical and clinical studies. In particular, metronomic cyclophosphamide treatment (140 mg/kg BW every 6 days) induces dramatic, near-complete regression of established xenografts of human U251 and rat 9L gliosarcomas in s.c. scid mouse models, whereas maximum tolerated dose cyclophosphamide effects only moderate tumor growth delays. Furthermore, metronomic cyclophosphamide-induced regression of 9L tumors is accompanied by reduced microvessel density, induction of thrombospondin-1 (TSP1) and impaired drug uptake, all of which are consistent with the prevailing view that anti-angiogenesis is a major factor in the therapeutic effectiveness of metronomic chemotherapy. However, VEGF receptor inhibition induces substantially greater anti-angiogenesis in these tumor models but does not induce tumor regression. Therefore, we sought to investigate other non-anti-angiogenic mechanisms to explain the substantial regression and have found that it is strongly associated with recruitment of the host innate immune system. In regressing 9L and U251 tumors, host (mouse) PEDF (pigment epithelium-derived factor; Serpinf1) and PAI-1 (plasminogen activation inhibitor-1; Serpine1) expression is increased, with the levels of PEDF RNA being strongly correlated (r=0.89) with TSP1 RNA in large sets of 9L and U251 tumors isolated various times after initiation of metronomic cyclophosphamide therapy. Notably, PAI-1 and TSP1 are both expressed by tumor-associated macrophages. Metronomic cyclophosphamide also increased Fas, which stimulates macrophage activation, and the macrophage marker CD68, supporting the involvement of tumor-associated macrophages. Fas is also associated with cytotoxic lymphocytes, such as NK cells, and NK cell markers and cytotoxic effectors were dramatically increased in metronomic cyclophosphamide-treated tumors. Furthermore, dendritic cell markers important for cell-target interactions and antigen presentation were increased by metronomic cyclophosphamide treatment. Cytokines and chemokines were also up regulated in the metronomic cyclophosphamide-treated tumors, suggesting that chemokine and cytokine gradients help mobilize the host innate immune system in regressing 9L and U251 tumors. These findings highlight the ability of metronomic chemotherapy to induce macrophage, natural killer cell, and dendritic cell recruitment, thereby activating innate immunity leading to regression of established gliosarcomas and potentially other tumor types. Supported by NIH grant CA49248 (to DJW).
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3813.
Abstract 3818: CD11c+CD123+ human myeloid dendritic cells generated using IL4, IL13 and IL3 show characteristics of myeloid derived suppressor cells
Authors: Edwards Jeremy V. , Zhang Hua , Mackall Crystal L. 
Published: February 2011

Abstract
Dendritic cells (DCs) are critical for initiation of adaptive immune responses, but they also play important roles in self-tolerance, and in tumor bearing hosts, some DC subsets may contribute to immune evasion. Myeloid derived suppressor cells (MDSC) are also important mediators of tumor immune evasion. The relationship between myeloid DC and MDSCs in humans remains unclear. Human myeloid DCs are defined as CD11c+CD123-, although some have reported dim CD123 expression on human myeloid DCs. We describe a CD11c+CD123+ myeloid DC subset that shows characteristics of a MDSC.
Monocytes elutriated from normal donors were incubated for 48hr with IL4 ± IL13 ± IL3. Monocytes cultured with all three cytokines acquired strong expression of both CD123 and CD11c, with 79.5% of cells CD123+CD11c+. CD11c+CD123+ cells demonstrated over-expression of both arginase and iNOS by flow cytometry and/or real-time PCR in comparison to myeloid derived DCs generated with IL4, GM-CSF and LPS (mean fluorescence intensity of arginase in the IL4, IL13 and IL3 generated cells of 5973 vs. 1961 in IL4, GM-CSF and LPS generated cells). CD11c+CD123+ DCs also coexpress IL-4Rα and IL7Rα, characteristics of MDSC and regulatory DC subsets respectively. IL3 appears to be a potent inducer of CD123 expression on CD11c+ DCs, since myeloid DCs generated with only IL4 and IL13, in combination or separately, showed a less pronounced phenotype (38-52.7% CD11c+CD123+). Moreover, CD123 expression on myeloid DCs appears to be a marker for arginase production since CD123+ cells within the IL4, IL13 and IL3 cultures show higher arginase expression than CD123- cells within the same cultures. Functional studies are ongoing.
In conclusion, human myeloid DCs generated by incubating monocytes with IL4, IL13 and IL3 have a novel CD11c+CD123+ phenotype and show characteristics of myeloid derived suppressor cells, raising the prospect that IL3 plays an important role in generating myeloid derived suppressor cells in humans.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3818.
Abstract 382: Platelets are implicated in the rapid host responses induced by cytotoxic chemotherapy which limit its anti-tumor efficacy
Authors: Milsom Chlöe C. , Shaked Yuval , Lee Christina R. , Man Shan , Kerbel Robert S. 
Published: February 2011

Abstract
Previous studies from our lab have shown that the overall anti-tumor impact of certain cytotoxic drugs such as paclitaxel (PTX) given at maximum tolerated doses (MTDs) or OXi-4503, a microtubule inhibiting vascular disrupting agent (VDA), can be reduced by a rapid systemic counter-acting host response (Y Shaked et al., Science 2006 and Cancer Cell 2008). The aforementioned host response includes rapid induction of elevated plasma levels of multiple cytokines or chemokines such as G-CSF and SDF-1, which can promote mobilization and tumor-homing/colonization of proangiogenic and tumor-growth promoting bone marrow-derived cells (BMDCs), including circulating endothelial progenitor cells. This reactive host response promotes recovery/repopulation of the drug treated tumor. Here we asked what might be the source(s) of the elevated levels of plasma chemokines and cytokines induced by MTD PTX. We found platelets - known major sources of SDF-1 and VEGF - become activated and release the contents of their alpha granules into the circulation (plasma), such that decreases in platelet associated SDF-1 (and VEGF) are observed accompanied by increases in their plasma levels. SDF-1 is known to mobilize a variety of BMDCs including CEPs. Because platelets are not known to be a source of G-CSF - which is also elevated in the plasma by MTD PTX - other cellular and/or non-cellular sources are likely affected as well by the drug treatment. We also found that pharmacologic-mediated reductions in platelet numbers or activation results in reduced expression of the counter-acting host cytokine/chemokine response induced by MTD PTX treatment, thus providing further support for a contribution of platelets. Finally we have found low-dose metronomic chemotherapy does not induce the counter acting host response which is likely due in part to a lack of effect on platelets. In summary, our results implicate a new way in which platelets can impact tumor biology and response to therapy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 382.
Abstract 3827: Interferons induce CXCR3-cognate chemokine production by melanoma
Authors: Lucht Amanda T. , Clancy-Thompson Eleanor , Dengel Lynn T. , Strieter Robert M. , Slingluff Craig L. , Mullins David W. 
Published: February 2011

Abstract
Immune-mediated cancer regression requires tumor infiltration by Ag-specific effector T cells, but lymphocytes are commonly sparse in melanoma metastases. Activated T cells express CXCR3, whose cognate chemokines are CXCL9/MIG, CXCL10/IP-10 and CXCL11/I-TAC. Little is known about expression of these chemokines in lymph node (LN) metastases of melanoma. We evaluated whether metastatic melanoma induces these CXCR3-cognate chemokines in human LN-derived tissues. Also, because these chemokines can be induced by interferon (IFN), we evaluated whether type I or II IFNs (IFN-α or IFN-γ, respectively) can modulate chemokine expression in an in vitro model of the human tumor microenvironment. Production of CXCL9-11 by melanoma-infiltrated nodes (MIN) was no different than tumor-free nodes (TFN); both produced less than activated LN (sentinel immunized nodes, SIN). These data suggest melanoma infiltration into LN neither induces nor reduces CXCL9-11. Stimulation with IFN-α or IFN-γ increased production of CXCL10-11 from MIN, but not TFN or SIN. IFN-γ also increased production of CXCL9 in MIN. In IFN-treated MIN, the primary source of CXCL9-11 was melanoma cells themselves, whereas CD14+ cells were the source of these molecules in SIN. Melanoma cell lines expressed IFN receptors and produced CXCL9-11 in response to IFN treatment. Treatment of melanoma cell lines or primary melanomas in vitro with IFN-γ induced the migration of CXCR3+ T cells in transwell assays, whereas T cell did not significantly migrate in response to resting melanoma cells. Thus, melanoma infiltration of LN is insufficient to induce the production of CXCL9-11, but melanoma may be a significant source of IFN-induced chemokines. Collectively, these data suggest that IFN-α or IFN-γ may act in the tumor microenvironment to increase the chemotactic gradient for CXCR3+ T cells, thus enhancing the capacity of therapeutic vaccines to mediate clinical efficacy.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3827.
Abstract 3833: Codon-optimized GM-CSF gene as an adjuvant to enhance the immunity of a human papillomavirus 16 E7 DNA vaccine in mice
Authors: Qiu Jian-Tai , Lin Cheng-Tao , Lai Chyong-Huey , Chang Kuo-Shiang , Wu Tzong-Shoon 
Published: February 2011

Abstract
Granulocyte macrophage-colony stimulating factor (GM-CSF) is a potent immunomodulatory cytokine. We have generated a novel codon optimized murine GM-CSF sequences as an adjuvant. The codon optimized murine GM-CSF plasmids significantly increased GM-CSF expression levels in all cells tested, including murine, monkey, and human cell lines. Here we show that coadministration of plasmids encoding the codon optimized GM-CSF sequence with the HPV-16 E7 DNA vaccine resulted in complete tumor regression in 100% of the C57BL/6 mice tested. Consistent with these findings, coadministarton of these plasmids elicited E7-specific CTL responses and a protective immune response against challenge of E7- expressing TC-1 tumors in mice. Histological analysis revealed that the tumor destruction occurred at the margins of the tumor, where there was a heavy lymphocytic infiltrate. We also created a TC-1 tumors expressing E7 and GM-CSF genes. Combining a GM-CSF secreting TC-1 vaccine with a systemic DNA vaccination successfully treated &gt;90% of the mice bearing an established tumor but showed no resistance to an unrelated ovarian cancer. This novel codon optimized GM-CSF plasmid may prove a practical molecular strategy for overcoming this limitation of DNA vaccines and potentiating immune responses to vaccines as well as other immunotherapeutic strategies.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3833.