bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1377: Vascular shutdown effects of tetra-arsenic oxide in solid murine tumors
Authors: Wen Lan Ying , Bae Su Mi , Kim Young Jae , Kim Kyung Tae , Ahn Woong Shick
Published: February 2011

Tetra-arsenic oxide [As4O6, 2,4,6,8,9,10-Hexaoxa-1,3,5,7-tetraarsatricyclo(3.3.1.13,7) decane]- one kind of arsenic compound, has shown more potent antitumor effect on cervix cancer cells. As4O6 also showed an inhibitory effect on tube formation of endothelial cells in vitro. The aim of this study was investigate the vascular shutdown effects of As4O6 on TC-1 cell implanted mice.

TC-1 cell, which is HPV 16 E6/E7 transfected cell, was used both in vitro and in vivo study. In vitro experiments: to assess cell viability MTT assay was employed, and morphology was determined by microscopy. In vivo experiments, 6 weeks old C57BL/6, female mice were used. 5′106 TC-1 cells were inoculated subcutaneously to abdomen and when tumor size reach to 350∼400 mm3 (12 to 14 days after inoculation when reaching a surface diameter of 8 to 10 mm), tumor bearing mice were randomly divided into two groups of PBS injected (n=14) and As4O6 (10 mg/kg b.w., i.p.) injected groups (n=14). Tumor volume was measured by caliper every two or three days for 3 weeks and recorded as volume diameter [longest surface length (a), width (b) and height (c), (a x b x c)]. Hoechst 33342 uptake method has been performed. One minute before harvest tumor tissue sections, Hoechst 33342 was injected and make frozen section. Tumor tissue were observed under 365nm microscope and photographed at x40. Each tumor image was constituted by 15∼35 pieces of view.

Through MTT assay, it was shown cell growth inhibitory effects of As4O6 on TC-1 cell lines. Cell proliferation reduced to 52.74%, when treat 4.00 uM As4O6, IC50 value was 4.05 uM after 24 hours treatment. There were also many changes on morphology after 24hours of As4O6 treatment, include: cell number was reduced, cell membrane became weak, and cytoplasm got scarce. In TC-1 cells challenged C57BL/6 mice, As4O6 delayed tumor growth. 3 weeks after As4O6 treatment, the tumor volume suppressed nearly 89.8% compared with control group (p<0.05). Hoechst 33342 can stain tumor area which has vasculature. We detected that Hoechst 33342 stained areas was significantly disappeared in the central of the tumor tissue from one hour after As4O6 injection. While non-treated tumor tissue was 100% stained, one hour treated As4O6 tumor tissue was stained only 12.5 %.

Conclusion: This study shows that As4O6 can make strong vascular shutdown effects on TC-1 cells implanted mice.

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 1377.
Abstract 1384: Prevention of angiogenesis by aldose reductase inhibition
Authors: Saxena Ashish , Tammali Ravinder , Reddy Aramati BM , Srivastava Satish K. , Ramana Kota V.
Published: February 2011

We have recently shown that Aldose Reductase (AR) known to be involved in oxidative stress-signaling, initiated by inflammatory markers, prevents human colon cancer cell growth in culture as well as in nude mice xenografts. Inhibition of AR also prevents azoxymethane-induced aberrant crypt foci formation in mice. In order to understand the chemopreventive mechanism of AR inhibition in colon cancer, we have investigated the role of AR in the mediation of angiogenic signals in vitro and in vivo models. Using human umbilical vein endothelial cells (HUVEC), we have investigated the effect of AR inhibition on VEGF- and bFGF- induced tube as well as spheroid formation in in vitro angiogenesis. We have also determined the invasion and migration of HUVEC by inhibition of AR. Further, to determine the in vivo efficacy of AR inhibition on angiogenesis, matrigel plug assay was performed using a rat model. Our results show that inhibition of AR significantly prevented the VEGF- and bFGF -induced proliferation and generation of reactive oxygen species in HUVEC. Further, AR inhibition also prevented the VEGF- and bFGF- induced secretion/expression of IL-6, MMP1, MMP9, ICAM, and VCAM. Matrigel plug model of angiogenesis in rats showed that inhibition of AR prevented the infiltration of blood cells, invasion, migration and formation of capillary like structures, and expression of blood vessels markers CD31 and vWF. Thus, our results demonstrate that AR inhibitors could be novel drugs for cancer chemoprevention by inhibiting angiogenesis.

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 1384.
Abstract 1386: A novel approach to develop anti-VEGFR2/KDR therapeutics: characterizing and targeting the G-quadruplex
Authors: Gaerig Vanessa C. , Fortin Jessica S. , Brown Robert V. , Brooks Tracy A.
Published: February 2011

The anti-angiogenic approach is traditionally considered for solid malignancies, but increasing evidence has also highlighted its involvement in the progression of hematological oncologies. Angiogenesis, including expression of key proteins, is aberrant in blood cancers such as leukemias, lymphomas, and myelomas, in addition to many solid malignancies, including pancreatic cancer. The most critical mediator of oncogenic angiogenesis is vascular endothelial growth factor (VEGF), which is recognized by two high-affinity receptor tyrosine kinases, VEGFR1 and VEGFR2 (KDR gene). VEGFR2 is necessary for the survival, growth, and differentiation of endothelial cells as its downstream effects include activation of the MAPK and the PI3K pathways, and its upregulation is observed under conditions of pathological angiogenesis. While VEGF/VEGFR2 signaling is known to act in a paracrine manner on endothelial cells, autocrine signaling has been observed in both malignant cells and hematopoietic stem cells. For endothelial and hematological malignancies, both autocrine and paracrine pathways can be targeted with anti-VEGFR2 therapies, potentially doubling the clinical efficacy. Intriguingly, the promoter region of KDR contains a GC-rich region of DNA within its the core promoter located −120 to −31 basepairs (bp) upstream of the transcriptional start site (TSS), putatively able to form several G-quadruplexes. These are unique DNA secondary structures that often serve as transcriptional silencer elements. In the present study, G-quadruplex formation has been isolated −80 to −38 bp upstream of the TSS, which contains five strings of guanines that form a mixed parallel/anti-parallel G-quadruplex, as determined by circular dichroism. The cleavage pattern of DMS footprinting and the major stop product of the polymerase stop assay confirmed that the dominant isoform is a parallel G-quadruplex occurring within the four 3’ runs of guanines. Mutant studies were performed to further determine guanines integral to G-quadruplex formation. An electrophoretic mobility shift assay was used to confirm the formation of a biologically relevant intramolecular structure. High-throughput screening identified a novel KDR-interactive agent, NSC643735, which thermally stabilized the G-quadruplex. This compound is currently being tested in vitro, and has high potential to transcriptionally downregulate KDR mRNA and, subsequently, VEGFR2 protein. These data characterizing the formation of a unique secondary structure in an important oncogene are promising to develop novel and specific anti-KDR therapy for the treatment of hematological and solid malignancies.

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 1386.
Abstract 1398: Aurora kinase A inhibition with MLN8054 radiosensitizes prostate cancer by inducing cell cycle arrest leading to increased prolonged DNA damage and apoptosis
Authors: Schleicher Stephen M. , Moretti Luigi , Hwang Misun , Kopsombut Prapaporn , Kim Kwang Woon , Spratt Daniel , Jaboin Jerry , Lu Bo
Published: February 2011

The purpose of this study is to investigate a potential new radioenhancer for the treatment of prostate cancer. Despite multiple promising treatment modalities, prostate cancer remains a significant cause of diminished quality of life and cancer death, indicating the need for more aggressive, targeted therapy. Aurora kinase A (AURKA) is a promising treatment target, but its inhibition has never been investigated in combination with radiotherapy in prostate cancer. Using the selective AURKA inhibitor, MLN8054, we performed clonogenic assay to investigate in vitro radiosensitization, cell cycle analysis to observe changes induced by AURKA inhibition, tumor growth delay to study in vivo effects, and histologic active caspase staining to measure apoptosis. We demonstrated radiosensitizing effects in vitro in PC3 and DU145 prostate cancer cell lines, which was confirmed in vivo by tumor growth delay in PC3 xenografts in nude mice. In addition, we investigated a potential mechanism for radiosensitization. We report that MLN8054 induces cell cycle arrest at G2/M and polyploidy in both prostate cancer cell lines, which is associated with sustained DNA damage from radiation as seen by increased immunofluorescence for γ-H2AX, a measure of DNA double-strand breaks. Staining for caspase-3 in xenograft samples showed increased apoptosis in irradiated samples pre-treated with MLN8054 compared to irradiation or inhibitor alone. Our findings support the potential use of AURKA inhibition with MLN8054 to enhance the effectiveness of radiation therapy for the treatment of prostate cancer, and suggest a potential mechanism underlying its radiosensitization effects.

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 1398.
Abstract 1400: Mitigation of radiation-induced injury with the nitric oxide synthase co-factor, tetrahydrobiopterin
Authors: Wang Jing , Lin Peck-Sun , Mikkelsen Ross B. , Gewirtz David , Anscher Mitchell S. , Graves Paul R.
Published: February 2011

The purpose of this study was to determine the effect of modulation of the nitric oxide (NO) and the cGMP/PKG signaling pathway on radiation-induced myofibroblast differentiation and radiation-induced injury in mice. Nitric oxide (NO) is a diverse second messenger critical for a variety of cell functions produced by nitric oxide synthases (NOS). NOS activity is regulated in part by the availability of co-factors including tetrahydrobiopterin (BH4). In the absence of BH4, NOS can become ‘uncoupled’ to produce superoxide anion which upon reaction with NO can form peroxynitrite, a highly reactive molecule known to contribute to protein tyrosine nitration. BH4 is approved for human consumption and therefore may represent a safe, viable strategy to reduce tissue injury from radiation. We show here that BH4 administration to mice prior to whole body irradiation with 9.5 Gy significantly increases mouse survival. In contrast, inhibition of NOS activity either before or after irradiation significantly reduced survival. We also show that the phosphodiesterase 5 inhibitor, sildenafil, inhibits the radiation-induced differentiation of human myofibroblasts. The mechanisms underlying the protective effects of BH4 and the role of NOS in radiation-induced tissue injury will be discussed.

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 1400.
Abstract 1404: Mitochondrial-targeted nitroxide JP4-039 ameliorates irradiation-induced delay in bone wound healing and inhibits orthotopic tumor growth
Authors: Rwigema Jean-Claude , Gokhale Abhay , Epperly Michael , Glowackie Julie , Wang Hong , Dixon Tracy , Patrene Ken , Wipf Peter , Greenberger Joel S.
Published: February 2011

In the case of a nuclear explosion, combined injuries will result from radiation dispersal resulting in burns and cellular damage as well as a concussion wave resulting in traumatic bone fractures. Currently no available drugs are available to protect against both irradiation damage and traumatic bone damage. A new mitochondrial targeted small molecule hemigramicidin S nitroxide JP4-039 has been developed and shown to protect against total body irradiation. JP4-039 was tested to determine its effects on irradiation-induced delay of osseous wound healing.

A model for osseous would healing was used where the right hind legs of C57BL/6NHsd female mice were irradiated to doses 0 to 30 Gy 24 hours prior to drilling tibial wounds. JP4-039 (10 mg/kg) was injected either 10 min before or after irradiation. Mice were sacrificed weekly after wounding; tibiae were removed and radiographed for measurement of hole diameter. The left non-irradiated leg served as a control for the irradiated right leg. To determine whether JP4-039 might enhance tumor growth while stimulating bone growth, we injected Lewis lung carcinoma cells subcutaneously above the knee in both legs. The tumor on the right leg was within the 10Gy irradiation field. Tumor growth was monitored daily with calipers. Tukey's test was used to assess statistical significance between control and treated groups.

Mice treated with JP4-039 before bone drilling showed significant improvement in osseous wound healing at 14 days and 21 days after drilling (0.55 ± 0.3 and 0.47 ± 0.04 mm, respectively), compared with 0.84 ± 0.05 and 0.64 ± 0.05 mm, respectively for control, untreated mice (p<0.05). Mice treated with JP4-039 prior to 20 Gy irradiation showed improved bone healing compared with controls at 21 days; Mice treated with JP4-039 after 20Gy showed improved bone healing at 21 and 28 days(p<0.05). Mice bearing orthotopic tumors treated with JP4-039 prior to irradiation and bone drilling showed significantly less increase in tumor volume compared with control group in both irradiated and non-irradiated legs (p < 0.05).

Thus, treatment with JP4-039 enhanced osseous bone healing when administered before wounding. Furthermore, JP4-039 enhanced healing when administered before or after irradiation. Finally, JP4-039 inhibited tumor growth and may have useful applications in cancer 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 1404.
Abstract 1406: Involvement of DNA repair pathways in DAG-lactone radiosensitization of human LNCaP cells
Authors: Galvin David J. , Thin Tin H. , Truman Jean-Paul , Shenker Larissa , Kolesnick Richard , Fuks Zvi , Haimovitz-Friedman Adriana
Published: February 2011

We previously demonstrated that pre-treatment of human androgen-dependent prostate cancer cell line LNCaP with either 12-0-tetradecanoylphorbol 13-acetate (TPA, a known protein kinase C activator), or diacylglycerol-lactone (DAG-lactone) radiosensitized these cells. This effect was mediated by down-regulation of ATM protein levels, thus de-repressing the enzyme ceramide synthase (CerS), ceramide generation and apoptosis. Here we assessed whether ATM down-regulation affected DNA repair pathways. LNCaP cells were pretreated for 16 hr with 10μM DAG-lactone and irradiated with 20 or 40Gy. Pre-treatment with DAG-lactone significantly enhanced apoptosis at 48 hours from 1.1% to 13.9% (20Gy) and 19.9% (40Gy) respectively. γ-H2AX and BRCA1 foci formation at 1 and 8 hours post-radiation were scored. Immunofluorescent staining for γ-H2AX demonstrated a dose-dependent increase in γ-H2AX foci formation at 20Gy and 40Gy for both 1hr and 8 hr time points. Our data also shows that 40Gy alone produces the same amount of DNA double strand breaks (dsbs; as measured by γ-H2AX at 8 hr) as 20Gy+DAG-lactone, yet 40Gy alone does not induce apoptosis in LNCaP cells, while 20Gy+DAG-lactone does, indicating that ATM down-regulation is necessary for the apoptotic response. Similarly there was a dose-dependent response in the generation of BRCA1 foci at 20Gy and 40 Gy at 8hr post-radiation. There was no difference in the BRCA1 foci between pre-treatment with DAG-lactone either at 20Gy or at 40 Gy at both 1 hr or at 8 hr post-radiation. While apoptotic degradation is associated with γ-H2AX phosphorylation, in LNCaP cells these events would be expected at 24-48 hr. Therefore the foci observed here are unlikely to be associated with apoptotic DNA degradation and most likely result from DNA dsbs. Our recent data showed that unrepaired DNA dsbs trigger post-transcriptional activation of CerS via a mechanism that remains unknown. Accordingly MRE 11, ATM or DNA-PKcs (SCID) deficiencies sensitize mice towards CerS-mediated apoptosis in crypt stem cells clonogens.

Although these studies indicate that HR is active, further experiments need to be carried out to explore whether non-homologous end-joining (NHEJ) is or is not functional in LNCaP treated with DAG-lactone and radiation. Hence, the trigger for CerS activation still needs to be established.

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 1406.
Abstract 1409: C20orf52 regulates cell death by ionizing radiation via modulation of ROS generation in the mitochondria
Authors: So Kwangsup , Yoo Young Do , Um Hong-Duck , Park Jong Kuk
Published: February 2011

It is known that ROS exerts an important role in various intracellular physiological phenomena. Mitochondria and its molecules are one of major intracellular ROS source. Generation of ROS in normal cell condition is mainly caused by intracellular metabolic processes, but several extra-cellular stresses including ionizing radiation (IR) also increase ROS production. In this study, we tried to identify new molecular target that regulate ROS generation after IR irradiation, and confirmed the C20orf52 (also named as MTMG and Romo-1) as one of the new candidate. In recent report, C20orf52 was identified a regulator of ROS generation in the mitochondria, and it is located at integral mitochondrial inner-membrane. We constructed C20orf52 over-expressing HEK293 cell, and then showed that this C20orf52 over-expressing cells have a radio-resistance via suppression of ROS production by IR. Down-regulation of C20orf52 by siRNA treatment in several cancer cells increased cell death by IR. C20orf52 over-expression also increased mitochondrial potential and JNK phosphorylation in MAPKinase pathway induced by IR. Therefore we suggest that C20orf52 might modulate ROS production by IR and regulates cell death.

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 1409.
Abstract 1423: SatB2, targeted by miR31, is a modulator in cancer microenvironment
Authors: Yu Xiang , Aprelikova Olga , Palla John , John Simone , Yi Ming , Stephens Robert , Risinger John I. , Jazaeri Amir , Niederhuber John
Published: February 2011

Cancer growth and metastasis involve an active interaction between primary tumor cells and the tumor's microenvironment, which consists of fibroblasts, vascular cells and inflammatory cells. Such interactions between the cancer and the cells within the microenvironment induce differential expression of genes that act to foster the growth, invasion and spread of tumor cells. This study utilizes pair-wise primary cultures of fibroblasts from normal and cancerous human endometrial tissues of the same patients to investigate the potential factors in the tumor microenvironment and their role in carcinogenesis. Both microRNA and mRNA profiling were performed on paired normal and cancer fibroblasts. SatB2 emerged as a promising candidate when its expression was shown to be significantly increased in fibroblasts surrounding the tumor tissues in all but one patients tested (n=8). Further investigation documented that SatB2 is a target for miR31, via 2 binding sites located within the SATB2 3’ UTR region. In concordance, miR31 is down-regulated in tumor fibroblasts, which further implicates the potential role of SatB2 in promoting a favorable environment for tumors. Ectopic expression of SATB2 in normal fibroblasts does not affect the growth rate of an endometrial cancer cell line (EC-1) in co-culture experiments; however, it did increase the motility of EC-1 cells towards these fibroblasts by multiple folds in both in vitro migration and invasion assays. A reduction of SATB2 expression in cancer activated fibroblasts (CAFs) by shRNA attenuated the migratory ability of EC-1 cells, further supporting the role of SatB2 in tumor aggression. Our observations also indicated that the conditioned media from CAFs were sufficient to induce the motility changes in the EC-1 cells. We believe the elucidation of SatB2 pathway(s) will assist in uncovering the intricate connections between tumors and their microenvironments, leading to new targets for treating tumor progression.

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 1423.
Abstract 1439: De novo expression of c-kit in prostate cancer cells interacting with the bone microenvironment
Authors: Zhe Xiaoning , Nabha Sanaa M. , Mukundan Anju , Giroux Craig N. , Cher Michael L. , Bonfil R. Daniel
Published: February 2011

Specific cellular and molecular interactions between prostate cancer (PC) cells and the bone microenvironment lead to expansion of the metastatic deposit, disease progression, and ultimately significant morbidity and mortality. We recently reported an association between expression of the tyrosine kinase receptor c-kit and clinical PC progression, with the highest expression seen in clinical metastatic bone specimens. Moreover, we found that intraosseous tumors formed by otherwise c-kit negative PC3 cells injected into human bone xenografts in SCID mice (SCID-hu model) expressed c-kit. To determine the relevance of these findings, we examined additional human PC cell lines, namely LNCaP, C42b, and DU145, for the expression of c-kit in vitro and in vivo. While none of the cultured PC cells expressed c-kit at the message or protein level, immunostaining for c-kit in intraosseous tumors produced by these cells in the SCID-hu model revealed a distinctive and specific reactivity in the cancer cell compartment. The induction of c-kit expression in PC cells growing within the bone microenvironment was emulated by co-culture of PC3 cells and bone-derived cells embedded in Matrigel, as demonstrated by RT-PCR analysis. In an attempt to mimic the de novo c-kit expression observed in the SCID-hu model and to identify potential signaling pathways triggered by c-kit activation by its ligand stem cell factor (SCF), normally expressed by the bone microenvironment, we transiently transfected PC3 cells with wild-type c-kit. Activation of c-kit was confirmed by immunoblotting for phosphorylated c-kit. Using the whole human genome Agilent microarray, we found that 220 genes were differentially expressed when SCF-treated and non-treated c-kit-transfected PC3 cells were compared, resulting in discrete functional sub-networks that were coordinately up or down-regulated. Most notably, pathway and genetic network analysis revealed that c-kit activation resulted in an up-regulated fibroblast growth factor-2-mediated sub-network. Taken together, these results suggest that c-kit induction in PC cells in the bone microenvironment modulates cell-cell interaction pathways and growth factor signaling systems that ultimately lead to expansion of bone metastasis.

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 1439.
Abstract 1440: Pediatric pulmonary microenvironment differs from adults: Implications for pulmonary metastasis
Authors: Hayes-Jordan Andrea Anita , Wang Yong Xin
Published: February 2011

Background: Ewing's sarcoma (ES) is the second most common bone tumor in children. Still, despite multimodal therapy, lung metastasis are the most common cause of demise. Older age has been found to portend a poorer survival in ES. Here we identify differences in the pulmonary micro-environment comparing pediatric to adult lung tissue.

Purpose: To identify molecular differences in the pediatric and adult lung micro-environment.

Methods: To evaluate whether adult lungs provide a more permissive metastatic environment compared to pediatric lungs, we measured the migration and invasion of ES cells in a modified Boyden chamber assay. ES TC-71 cells, know to have moderate metastatic potential and ES LM-7 cells which are highly metastatic, were compared normal lung epithelial cells (CRL2300). Lungs from 4 week old adolescent and 9 week old adult nude mice, were morselated and placed in a modified Boyden chamber. To compare expression of metastatic genes in pediatric compared to adult lung, RNA was extracted from 4 week and 9 week nude mouse lung and a focused microarray of 128 genes were evaluated for differential expression.

Results: The TC-71 cell line and the CRL2300 cells had equal area of migration to adolescent and adult lung. Whereas, LM-7 had a significantly larger area of cell migration to the adult lung compared to the adolescent lung.(p=0.033) The focused gene expression profile demonstrated significant differential expression of 2 genes, Interleukin 1-beta (IL1-B), and Integrin beta-3 (ItgB3). IL1-B was significantly under-expressed and ItgB3 was significantly over-expressed in adolescent lung tissue compared to adult lung tissue.

Conclusions: Highly metastatic ES LM-7 cells have a significantly decreased migration to the adolescent/pediatric lung compared to adult lung tissue. This implies a different microenvironment in adolescent lung which does not have the same affinity for metastatic Ewing's sarcoma cells as the adult lung. Furthermore, differential gene expression of IL1-B and ItgB3 and may contribute to the cause of this difference since ItgB3 regulates cell adhesion, invasion and low doses of IL1-B can cause tumor cell eradication.

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 1440.
Abstract 1448: Novel ALK5 inhibitor as an anti-metastasis agent in both breast cancer cells in vitro and MMTV/cNeu transgenic mice in vivo
Authors: Park Chul-Yong , Kang Bo M. , Kwon In H. , Moon So J. , Son Jee Y. , Yoon Eun J. , Kim Dae-Kee , Sheen Yhun Y.
Published: February 2011

We have synthesized novel ALK5 inhibitors and investigated their effects of on breast cancer cells as well as tumor bearing MMTV/cNeu transgenic mice and orthotopically injected mice. The treatment of ALK5 inhibitors into breast tumor bearing MMTV/cNeu transgenic mice and orthotopically injected mice inhibited lung metastasis compared to that of untreated animal. The level of ß-casein in the lung of the breast tumor bearing MMTV/cNeu transgenic mice and orthotopically injected mice were decreased by ALK5 inhibitor treatment. The levels of CXCR4, MMP2 and MMP9 were decreased in the lung and breast tumor of ALK5 inhibitors treated breast tumor bearing MMTV/cNeu transgenic mice and orthotopically injected mice. The action mechanisms of new ALK5 inhibitors were investigated in both breast cancer cells and breast cancer mice model. TGFß signaling, phosphorylated smad2/3 based from the western blot analysis in vitro and in vivo as well as confocal microscopic analysis in vitro was decreased by ALK5 inhibitors. And also, results from wound healing assay and invasion assay showed strong inhibition of TGFß action by new ALK5 inhibitors. This data strongly argue that ALK5 inhibitors have anti-metastasis effects via inhibiting ALK5 activity in vivo and in vitro. In order to gain further insight into the mechanism of ALK5 inhibitor, we have carried out experiment to examine the effects of ALK5 inhibitor in human breast cancer MDA-MB-231 cell. TGFß stimulated mesenchymal markers of epithelial mesenchymal transition (EMT), such as Snail, TWIST, N-cadherin and these were inhibited by ALK5 inhibitor concomitant treatment. Also, TGFß increased tumor invasion markers such as MMP2, and MMP9 which were inhibited by ALK5 inhibitor concomitant treatment. In addition, ALK5 inhibitors recover the level of E-cadherin, SnoN, Ski which were downregulated by TGFß treatment. In summary, ALK5 inhibitors have anti-metastatic effect on human breast cancer cells as well as tumor bearing MMTV/cNeu transgenic mice.

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 1448.
Abstract 1449: Chronic consumption of a high fat diet increases solid tumor growth and lung metastasis in BALB/c mice subcutaneously injected with CT26 colon cancer cells
Authors: Park Heesook , Song Hyerim , Park Jung Han Yoon
Published: February 2011

Epidemiological and laboratory animal studies suggest that dietary factors in general and specifically high dietary fat and calories increase the risk of colon cancer. In the present study, we determined whether dietary fat increases solid tumor growth and metastasis of colon cancer using a CT26 mouse colon cancer xenograft model. Four-week old, male BALB/c mice was fed a purified diet containing 34.9% (w/w) fat (high fat diet, HFD), or 4.3% fat (low fat diet, LFD) for a period of 16 weeks. After 16 weeks, CT26 colon cancer cells were injected subcutaneously into the animals’ right flank. 5 weeks after cell injection, animals were killed. The mean body weight was slightly increased in the HFD group (33.9 ± 0.46 g) as compared to the LFD group (31.9 ± 0.37 g). The weights of the liver, lung, and epididymal fat pad were also increased in animals fed the high fat diet. Solid tumor growth (tumor volumes and weights) and the number and volume of tumor nodules in the lung were markedly increased in the HFD group as compared to the LFD group. Immunohistochemical staining of tumor tissues revealed that the expression of proliferating cell nuclear antigen (PCNA), platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31, an angiogenesis marker), vascular endothelial growth factor, and phospho-tyrosine phosphatase (CD 45, a monocytes/macrophage marker) was markedly increased in the HFD group as compared to those in the LFD group. Results from cytokine array revealed that the expression of IFN-γ, IL-1ra, IL-16, CXCL9, MIP-2, CCL5, and TNF-α was up-regulated in the sera of the HFD group as compared to the LFD group. These results indicate that increases in angiogenesis, infiltration of monocytes and cytokine expression probably contribute to increased tumor growth and lung metastasis in mice fed the high fat diet.

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 1449.
Abstract 1453: Effect of systemic administration of chromogranin A on tumor metastatization in animal models
Authors: Dondossola Eleonora , Crippa Luca , Colombo Barbara , Gasparri Anna , Curnis Flavio , Corti Angelo
Published: February 2011

Chromogranin A (CgA), an acidic glycoprotein present within secretory vesicles of many normal and neoplastic endocrine and neuroendocrine cells, is abnormally released in circulation in patients with neuroendocrine tumors. We have previously shown that local production of CgA can affect tumor growth by affecting the tumor microenvironment. Furthermore, we have recently shown that circulating CgA is increased also in a subpopulation of patients with non small cell lung cancer (NSCLC), with important prognostic implications. The present study was undertaken to investigate the role of CgA in tumor metastatization. We have found that systemic administration of CgA to mice can inhibit the number of lung metastasis formed by B16F10 mouse melanoma and by TS/A mouse mammary adenocarcinoma cells, when these cells were injected into the tail vein. The dose-response curve was bell-shaped, maximal effects being obtained with a dose of 0.2 ug/mouse administered daily (i.v for four days after tumor cell injection). Remarkably, this dose produced circulating levels similar to those found in NSCLC patients. The CgA1-78 N-terminal fragment (called vasostatin-1, VS-1) was sufficient to exert anti-metastatic effects in both models, suggesting that the N-terminal region of CgA contains a bioactive site. CgA could also inhibit B16F10 cell migration and invasion in vitro, and vessel formation in different in vitro and in vivo angiogenic assays. These results suggest that abnormal levels of circulating CgA in cancer patients can contribute to regulate tumor metastatization, possibly by affecting different steps of the metastatic cascade.

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 1453.
Abstract 1454: Telomerase-specific adenoviral labeling and treatment of hepatocellular carcinoma
Authors: Kishimoto Hiroyuki , Urata Yasuo , Fujiwara Toshiyoshi , Bouvet Michael , Hoffman Robert M.
Published: February 2011

We previously constructed telomerase-specific replication-selective adenoviruses OBP-301 (Telomelysin) and OBP-401 which contains the GFP gene (Telomelysin-GFP, TelomeScan). The replication of these adenoviruses is regulated by a human telomerase transcriptase (hTERT) promoter and allows specific targeting and labeling of tumor cells in vivo (Proc. Natl. Acad. Sci. USA, 1-6, 14514-14517, 2009 and Molecular Cancer Therapeutics 8, 3001-3008, 2009). The aim of this study was to validate systemic injection of these telomerase-specific replicating adenoviruses for in vivo hepatocellular carcinoma (HCC) GFP labeling and tumor-cell killing. We assessed the antitumor efficacy and the tumor labeling ability of the adenoviruses in an orthotopic mouse model using the Hep3B-GFP human HCC cell line. Biweekly i.v. treatment of OBP-301 caused a significant inhibition in liver tumor growth and showed a reduction in the number of liver tumor nodules compared to controls. To assess the GFP tumor labeling ability of OBP-401 for HCC, an orthotopic liver tumor model of unlabeled Hep3B was used. After i.v. injection of OBP-401, the liver tumors were labeled with GFP and visualized by fluorescence imaging. These results suggest that the two viruses, OBP-301 and OBP-401, when administrated systemically, are novel anticancer agents and labeling tools for HCC, respectively, with potential for human use.

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 1454.
Abstract 1457: Intraoperative contrast-enhanced ultrasonography using perflubutane for liver resection
Authors: Uetake Hiroyuki , Ishikawa Toshiaki , Mitsunori Yusuke , Tanaka Shinji , Arii Shigeki , Sugihara Kenichi
Published: February 2011

[Background] Bevacizumab (Bmab) is now widely used for metastatic colorectal cancer (CRC) treatment in the world. Through the high response of metastatic CRC to Bmab combination therapy, neoadjuvant chemotherapy using Bmab plus FOLFOX is a new strategy for the treatment of liver metastasis. As liver metastases shows high shrinkage by Bmab plus FOLFOX therapy, intraoperative contrast-enhanced ultrasonography using perflubutane (SONAZOIDTM) is supposed to be effective for complete detection and resection of small metastatic lesion after chemotherapy. [Material and methods] Forty-five patients with metastatic CRC were underwent Bmab combination chemotherapy in our institute (response rate and disease control rate was 72% and 94%, respectively). Among them, 8 patient (18%) were underwent liver resection after Bmab+mFOLFOX6 treatment. Median cycle of preoperative chemotherapy was 6.5 (5-17). [Result] By CT angiography, 52 liver metastases were detected before chemotherapy and 27 after chemotherapy. By intraoperative contrast-enhanced ultrasonography using SONAZOIDTM, 31 liver tumors were detected and resected. In histological examination, 13 of the resected 31 lesion (41%) were without viable cancer cell (pCR). [Conclusion] Intraoperative contrast-enhanced ultrasonography was effective for complete resection of liver metastasis after Bmab plus FOLFOX therapy. High pCR rate for liver metastasis was observed by Bmab plus FOLFOX 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 1457.
Abstract 1459: Proteomic identification of a novel antitumor protein from myeloid-derived suppressor cells in breast cancer
Authors: Boutte Angela , Lin Charles
Published: February 2011

Myeloid derived suppressor cells (MDSCs) are significantly increased in hematopoietic organs of cancer patients and animals bearing tumors. The number of MDSCs is directly proportional to tumor malignancy. MDSCs infiltrate into tumors and facilitate tumor progression and metastasis. In searching for molecular mediators responsible for tumor metastasis, we used proteomics and mass spectrometry in combination with a mammary tumor model that includes two distinct tumor cell lines, 67NR and 4T1, which are derived from a single mammary tumor. While these cell lines form primary tumors with equivalent growth kinetics, they differ dramatically in their metastatic potential. MDSCs were isolated from spleens of tumor-bearing mice and compared for protein expression profiles using 2D-DIGE-MS. There was a significant reduction of a novel protein, neutrophilic granule protein (NGP), in MDSCs from 4T1- compared to 67NR- tumor-bearing mice. Further analysis showed that NGP was expressed in myeloid cells, but not in epithelial tumor cells. Addition of 4T1 tumor cell conditioned media decreased the gene expression in promyelocyte cells. Computational analysis of NGP indicates presence of a putative cystatin domain, a known cathepsin inhibitor site. Indeed, forced expression of NGP in cultured cells inhibited endogenous cathepsin B activity. Since cathepsins have been linked to tumor invasion and metastasis, we postulated that NGP has an anti-tumor role. To test this hypothesis, we generated 4T1 cells stably expressing NGP (4T1-NGP) or empty vector (4T1-Vec), and found that expression of NGP in tumor cells inhibited Matrigel invasion, but not cell proliferation. Furthermore, implantation of 4T1-NGP cells in mammary pad of syngeneic Balb/c mice resulted in a significant reduction of primary tumor growth and lung metastasis. Histological examination reveals reduced tumor angiogenesis and increased apoptosis. Collectively, this study identified a novel protein, NGP, in MDSCs acts as a negative mediator of tumor growth and metastasis, likely through inhibition of cathepsin activity. Down-regulation of NGP in the tumor microenvironment contributes to tumor growth and metastasis. NGP may act as a novel protease inhibitor and may be a therapeutic target for breast cancer.

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 1459.
Abstract 1465: Histone deacetylase inhibitor suberoylanilide hydroxamic acid blocks epithelial-mesenchymal transition exhibited by Tera-2 embryonal carcinoma cells with a stem cell phenotype
Authors: Chu Khoi , Player Audrey N. , Pickei Ross , Robertson Fredika M.
Published: February 2011

The process of epithelial-mesenchymal transition (EMT) is associated with tumor progression. Tumor cells that undergo EMT have altered morphology and acquire a stem cell phenotype. Although there are numerous studies that demonstrate that shRNA approaches targeting specific transcription factors can block EMT, there are few pharmacological agents yet identified that can alter the process of EMT. Recent evidence suggests that the silencing of E-cadherin during EMT is dependent upon the activity of specific histone deacetylyase enzymes, which are known to regulate gene transcription. We used Tera-2 (HTB 106) embryonal carcinoma cells grown as 3 dimensional spheroids as a model of a tumor type characterized by a cancer stem cell phenotype. We observed that the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA; vorinostat; Zolinza®, Merck), which is FDA approved for treatment of cutaneous T cell lymphoma, significantly inhibited expression of over 55 distinct genes associated with EMT including those regulating responses to growth factors, adhesion, motility, and genes associated with maintenance/survival of stem cells, including Alpha 5 Integrin/fibronectin receptor alpha (ITGA5), Caldesmon 1 (CALD1), Epidermal growth factor receptor (EGFR), Forkhead box C2 (MFH-1, mesenchyme forkhead 1) (FOXC2), Glycogen synthase kinase 3 beta (GSK3 beta), Jagged 1 (JAG1), Integrin-linked kinase (ILK), Matrix metallopeptidase 2 (MMP2), Notch 1 homolog (NOTCH1), Platelet-derived growth factor receptor beta (PDGFb), Snail homolog 1 (SNAI1), Snail homolog 2 (SNAI2), Transcription factor 4 (TCF4), transforming growth factorβ1 (TGFβ1), Twist homolog 1 (Twist1), Wingless-type MMTC integration site family, member 5b (WNT5b), and Zinc finger E-box binding homeobox 2 (ZEB2). SAHA also altered the morphology of Tera-2 cells when cultured as adherent cells, which was associated with inhibited of both invasion, and anchorage independent growth in soft agar. In addition, SAHA inhibited the self-renewal capacity of Tera-2 tumor spheroids, which was associated with changes in expression of stem cell surface markers. Studies are ongoing to identify the HDAC enzymes that regulate expression of genes associated with the process of EMT and maintenance/survival of Tera-2 tumor spheroids that are targeted by SAHA. These studies suggest that HDAC inhibitors may be promising agents to reverse the process of EMT and may be potentially effective as inhibitors of tumor progression.

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 1465.
Abstract 1468: Antagonism of platelet-derived growth factor receptor-α as a potential therapeutic for prostate cancer bone metastases
Authors: Russell Mike R. , Liu Qingxin , Lei Hetian , Kazlauskas Andrius , Fatatis Alessandro
Published: February 2011

Prostate adenocarcinoma is the second leading cause of cancer death among men, due primarily to the fact that the majority of prostate cancers will eventually spread to the skeleton. Metastatic dissemination requires a complex series of coordinated events that result in cells that escape from the primary tumor into the circulation and eventually colonize distant organs. The ability of disseminated cancer cells to evolve into metastases of clinically significant size depends strongly on their compatibility with, and ability to utilize, this new micro-environment. We previously showed that bone-metastatic prostate cancer cells exposed to human bone marrow respond by activation of cell survival pathways, such as PI3K/Akt., and that these events are mediated by the alpha-receptor for platelet-derived growth factor (PDGFRα). We also have shown that prophylactic antagonism of this receptor using IMC-3G3, a monoclonal, humanized antibody currently in phase II trials, was able to significantly reduce tumor burden in an in vivo model of skeletal metastasis.

Here we show that IMC-3G3 can be used in a curative manner to reduce the growth of previously established skeletal lesions. In addition, we have successfully combined this antibody with zoledronic acid, a bisphosphonate commonly used to treat skeletal metastases, with encouraging results. Interestingly, treatment with a monoclonal antibody targeting only host PDGFRα failed to show any significant difference over saline treated mice.

Lastly, we sought to dissect the mechanism by which PDGFRα may exert its role within the bone microenvironment. We provide evidence that soluble components of human bone marrow can activate PDGFRα through a mechanism that does not require the canonical binding of PDGF ligand(s) to the receptor. To establish the relevance of this phenomenon in vivo, we employed a PDGFRα mutant lacking the extracellular ligand-binding domain. Our studies show that this truncated PDGFRα was able to restore bone-metastatic potential of prostate cancer cells as effectively as the full-length form of the receptor. In addition, we have found that expression of PDGFRα, and its related signaling events, results in the regulation of a specific set of genes that we believe may aid in the survival and growth of metastatic prostate cancer cells within the bone. Studies to understand the contribution of several of these putative bone metastatic genes are currently underway in our laboratory.

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 1468.