bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1819: Fecal RNA test using microRNA expressions of exfoliated colonocytes for colorectal cancer screening
Authors: Koga Yoshikatsu , Yasunaga Masahiro , Takahashi Amane , Kuroda Junichiro , Matsumura Yasuhiro
Published: June 2012

Background & Aims: To reduce the mortality rate of colorectal cancer (CRC), the development of a screening test for its early diagnosis is needed. We have been developing new CRC screening methods by applying molecular biological tools to exfoliated colonocytes. As part of this development, we have reported in the last few years the fecal RNA test using CRC-related gene expression analysis for CRC screening. In this study, we conducted microRNA (miRNA) expression analysis in exfoliated colonocytes isolated from feces for CRC screening.

Methods: We initially analyzed CRC-related oncogenic miRNAs (miR-17-92, miR-21, and miR-135) in CRC tissue and exfoliated colonocytes. We subsequently selected other oncogenic miRNAs for CRC detection using TaqMan MicroRNA Array v3.0 (corresponds to miRBase release 14). We finally analyzed all oncogenic miRNA expressions in colonocytes isolated from feces from 197 CRC patients and 119 healthy volunteers by quantitative real-time PCR.

Results: miR-17-92, miR-21, and miR-135 showed a significantly higher expression in CRC tissue than in normal tissue. The overall sensitivity of miRNA expression analysis in the exfoliated colonocytes was 74.1% (146/197, 95% CI; 67.4-80.1) and the specificity was 79.0% (94/119, 95% CI; 70.6-85.9). Because the sensitivity and specificity using CRC-related oncogenic miRNA expressions were relatively low, we selected 12 miRNAs as candidates of CRC detection using TaqMan MicroRNA Array v3.0. Conclusions: We have indentified suitable miRNAs for CRC detection in exfoliated colonocytes. Additionally, we report the overall sensitivity and specificity of miRNA expression analysis in exfoliated colonocytes using these oncogenic miRNAs in this annual meeting. miRNA expression analysis of colonocytes isolated from feces may be useful for CRC screening.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1819. doi:10.1158/1538-7445.AM2011-1819
Abstract 1821: Marital status and adherence to scheduled free breast cancer screening among uninsured inner city women
Authors: Spencer Cherie , Higginbotham Kimberly , Williams Carla , Miller Kim , Lea Kelda , Anders Kyle B. , Frederick Wayne A.I. , Laiyemo Adeyinka O.
Published: June 2012

Background: Mammography screening is associated with reduction in mortality from breast cancer. Women with lower socioeconomic status have increased burden from breast cancer which may be attributable to lower access and uptake of screening. The impact of family support in promoting uptake of breast cancer screening among women with low socioeconomic status is not well understood.

Aim: We sought to examine whether marital status influences adherence to scheduled free breast cancer screening with clinical breast examination and mammography among uninsured inner city women.

Methods: Howard University Cancer Center offered free breast cancer screening (clinical breast examination and mammogram) for uninsured women who are aged 40 and older in the Washington DC area. Participants were referred from community health organizations’ outreach programs in Maryland, District of Columbia and Maryland. Our analytic cohort comprises patients who were scheduled for free breast cancer screening from July 2009 to June 2010. The participants were categorized as non-adherent (no shows) if they did not cancel their appointments and did not show up for the breast cancer screening. We used Poisson regression models to evaluate the association of race with adherence to the scheduled breast cancer screening and adjusted for potential confounders. We calculated Relative Risks (RR) for no shows and 95% Confidence Intervals (CI).

Results: A total of 460 patients were scheduled for screening. The mean age was 49.7 years; 195 (43.4%) were Hispanic and 215 (47.9%) were black. 186 (41.3%) patients were married and 264 (58.7%) were not. Married patients were less likely to be born in the United States (6% vs 21%; P < 0.0001) but were comparable in age with unmarried patients (mean age = 49.4 years vs 50.4 years; P = 0.239). 345 (75%) patients showed up but 115 (25%) were non-adherent for their scheduled appointment. When compared with unmarried patients, married patients were less likely to be non-adherent to their breast cancer screening appointment, adjusted RR = 0.58; 95%CI: 0.40 – 0.84, P value = 0.004.

Conclusions: Although a quarter of uninsured women are non-adherent for their free breast cancer screening appointments, married women are less likely to be non-adherent. This suggests that educational outreach programs targeting men may be a potential mechanism to increase uptake of breast cancer screening among underserved women.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1821. doi:10.1158/1538-7445.AM2011-1821
Abstract 1824: Effects of high-fat diet feeding on solid tumor growth and lung metastasis in the Lewis lung cancer allograft model
Authors: Song Hyerim , Park Heesook , Park Jung Han Yoon
Published: June 2012

Epidemiological studies indicate that overweight and obesity are associated with increased risk of developing several cancers. In the present study, we determined whether a chronic consumption of a high-fat diet increases solid tumor growth and metastasis in the C57BL/6N mouse lung cancer allograft model. Four-week old, male C57BL/6N mice were fed a purified diet containing 34.9% (w/w) fat (high fat diet) or 4.3 % fat (control diet) for 16 weeks, and then Lewis lung carcinoma cells were subcutaneously injected at 5 × 104 cells per mouse into the animals’ left flank. The primary tumor was resected 3 weeks later, and all animals were killed 21 days after the surgery. The high-fat diet feeding significantly increased the body weights and energy intakes as compared with those in the control diet group. The weights of the liver, spleen, epididymal fat pad, and mesenteric fat pad were increased in the high-fat diet group compared to the control group. The high fat diet increased solid tumor growth as well as the number and the size of colonies in the lung. Immunohistochemical staining of tumor tissues revealed that the expression of Ki67 (prototypic cell cycle related nuclear antigen), CD45 (phospho-tyrosine phosphatase), CD31 (platelet endothelial cell adhesion molecule-1), vascular endothelial growth factor (VEGF), cyclin-dependent kinase 4, cyclin A, and cyclin D1 was significantly increased in the high-fat diet group compared to the control group. The high-fat diet feeding significantly increased the blood levels of leptin, monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), granulocyte-colony stimulating factor (G-CSF), triggering receptor expressed on myeloid cells-1 (TREM-1), angiopoietin-1 (Ang-1), chemokine ligand 16 (CXCL16), plasminogen activator inhibitor-1 (PAI-1), and stromal cell-derived factor-1 (SDF-1) compared to the control group. Results of the present study indicate that the prolonged consumption of a high-fat diet enhances the secretion of cytokines, chemokines, matrix metalloproteinases, and angiogenesis-related proteins and increases the infiltration of macrophages into tumor tissues which, in turn, stimulate the proliferation and metastasis of lung cancer cells. The increased angiogenesis probably contributes to increased tumor growth and metastasis in mice fed on a high-fat diet.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1824. doi:10.1158/1538-7445.AM2011-1824
Abstract 1828: Omega-3 fatty acids delay the growth of mammary tumors in MMTV-Erbb2 mice
Authors: Wang Jing , Zhang Chunlan , Deng Guorui , Chen Yong , Berquin Isabelle
Published: June 2012

Purpose: Dietary fats have long been suspected of contributing to breast cancer risk, but the effects of various fatty acids are still not clear. We used MMTV-Erbb2 transgenic mice to determine how different fatty acids influence mammary tumor development and what the possible mechanisms are.

Methods and Results: Female MMTV-Erbb2 transgenic mice (FVB-Tg(MMTV-Erbb2)NK1Mul/J, n=13-14 per group) were placed on defined diets with 30% calories from fat, enriched in different types of fatty acids: saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), omega-3 polyunsaturated fatty acids (n-3 PUFA), omega-6 polyunsaturated fatty acids (n-6 PUFA) and control diet with a nutritionally balanced mixture of the above fatty acids. The results showed that the incidence of palpable mammary tumors, their multiplicity and average tumor weight were significantly lower in the n-3 PUFA enriched diet group than in other diet groups (p<0.05, ANOVA). Moreover, the tumors from the n-3 PUFA diet group showed decreased cell proliferation and increased apoptosis (as determined by immunohistochemical staining for Ki67 and cleaved caspase3, respectively) compared to other groups. However, there were no significant differences in the number of hyperplasic nodules detected in mammary glands by carmine alum whole mount staining. SFA, MUFA and n-6 PUFA were not statistically different from control.

Conclusions: Our results suggest that n-3 PUFA can inhibit HER-2/neu-induced mammary tumor growth in this mouse model, but that n-3 PUFA has no effect on the very early stage of tumorigenesis. The mechanism of the antitumor effects of n-3 PUFA may be related to inhibition of cell proliferation and promotion of apoptosis.

This project was supported by an Intramural Research Support Grant (Kulynych Fund) entitled: “Pilot study on the effects of different dietary fatty acids on mammary cancer risk”.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1828. doi:10.1158/1538-7445.AM2011-1828
Abstract 1837: Clinical diversity and tumor spectrum in Xeroderma Pigmentosum Brazilian patients
Authors: Santiago Karina M. , Maschietto Mariana , Vairo Filippo Pinto , Ashton-Prolla Patricia , Medeiros Paula , Rogatto Silvia Regina , Achatz Maria Isabel Waddington
Published: June 2012

Xeroderma Pigmentosum (XP) is a rare autossomic recessive hereditary disorder in which normal ability to repair DNA damage caused by ultraviolet light (UV) is deficient due to the presence of mutations in nucleotide excision repair pathway genes (XPA, ERCC3, XPC, ERCC2, DDB, ERCC4 and ERCC5) or in POLH gene, responsible for the replication of damaged DNA on the leading strand. These different genetic alteration result in distict phenotypes classifified into eight genetic complementation sub groups, XP-A to XP-G and a variant group XP-V. Clinical characteristics of the syndrome include skin poikiloderma and eye hypersensibility to UV radiation. Carriers have an increased risk of early onset multiple cutaneous, mucocutaneous and ophthalmologic malignancies when compared with normal population. Occasionally neurological impairment is observed in certain variations of the syndrome. Given the paucity of specific literature data about the incidence and clinical profile of the disease in Brazilian population, this study was conducted to describe clinical diversity and tumor spectrum of XP syndrome in Brazilian population. Twenty-nine XP patients from 19 non-related families from Hospital A.C. Camargo (São Paulo, Brazil), Hospital Universitário Alcides Carneiro (Campina Grande, Paraíba) and Hospital de Clínicas de Porto Alegre (Rio Grande do Sul, Brazil). Data collection was performed after genetic counseling and signed informed consent. Clinical characteristics were obtained through medical ascertainment. The median age was 24 (4 to 61 years) in a predominant female population (19/29). Basal cell carcinomas (BCC) were the most frequent neoplasm, present in 86,20% (25/29) of all patients, with the occurrence of up to 60 BCCs in a same patient. Squamous cell carcinomas and melanoma were present in 66,7% and 37,5% respectively. Six patients developed the first malignant tumor before the age of six and four patients developed the first tumor in early adulthood. One male patient developed a diffuse gastric cancer at 50 years old. Visual impairment occurred in 45,83% of the patient. Despite the unknown frequency of affected patients, the Brazilian population of XP cases seems to be underestimated. Genetic counseling is fundamental to provide information to carriers and family members and enable preventive measurements.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1837. doi:10.1158/1538-7445.AM2011-1837
Abstract 1841: Dietary morin suppresses colitis-associated colorectal carcinogenesis in rats
Authors: Tanaka Takuji , Yasui Yumiko , Terasaki Masaru , Masuda Sonoko
Published: June 2012

A flavonol, morin, found in almonds, mill, fig and other moraceae which are used as dietary agents and also as herbal medicine. Morin acts as a potent antioxidant, xanthine oxidase inhibitor, PKC inhibitor, cell proliferation inhibitor, apoptosis inducer, modulator of LOX and COX activities. Based on our previous studies on chemopreventive ability of morin in tongue and colon carcinogenesis (Carcinogenesis 20: 1477-84, 1999; Int J Cancer 83: 381-6, 1999; Oncol Rep 6:1333-40, 1999), we aimed to determine possible inhibitory potential of this compound in colitis-associated colon carcinogenesis in rats initiated with AOM and promoted by DSS. Male F344 rats initiated with a single s.c. injection of AOM (20 mg/kg bw) were promoted by 1.5% DSS in their drinking water for 7 days. They were then given a basal diet containing 50, 250 and 1000 ppm of morin for 17 weeks. At wk 20, the development of colonic adenocarcinoma was significantly inhibited by feeding with morin at a dose level of 1000 ppm (45% incidence, p=0.0141; and 0.73 +/- 1.01 multiplicity, p<0.05), when compared to the AOM/DSS group (93% incidence and 2.36 +/- 1.95 multiplicity). The incidence and multiplicity of adenocarcinomas in the rats received 50 ppm (60% incidence and 1.20 +/- 1.23 multiplicity) and 250 ppm (64% incidence and 1.09 +/- 1.04 multiplicity) of morin in diet were also smaller than that of the AOM and DSS group, but the differences were insignificant. The treatments inhibited colonic inflammation, and also modulated proliferation, apoptosis, and expression of several pro-inflammatory cytokines, such as NF-kappaB, Stat3, TNF-alpha and IL-1beta in the adenocarcinomas. Feeding with morin lowered tissue polyamine levels of colonic mucosa. Our findings indicate that dietary morin is able to inhibit colitis-related colon carcinogenesis in rats. Also, our model provides knowledge applicable to prevent inflammation-associated colon cancer in humans.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1841. doi:10.1158/1538-7445.AM2011-1841
Abstract 2148: Thalidomide and liposomal doxorubicin inhibit differentiation and function of human osteoclasts
Authors: Chen Yu-Jen , Chen Yu-Yawn , Chi Chih-Wen , Liao Hui-Fen
Published: June 2012

Background: Thalidomide inhibits angiogenesis and growth of multiple myeloma. Liposomal doxorubicin is mainly absorbed and degraded by monocyte-macrophage lineage in vivo, implicating a possible targeting effect. Given that osteoclasts (OCs) are derived from precursors of monocyte-macrophage lineage, we hypothesized that thalidomide, liposomal doxorubicin and their combination may affect osteoclastogenesis.

Materials and Methods: We isolated CD14+ monocytes from peripheral blood mononuclear cells of healthy subjects and generated OCs under stimulation with macrophage-colony forming factor and receptor activator of NK-κB ligand. Cell viability, surface CD51/61 expression, and tartrate-resistant acid phosphatase (TRAP) activity were assessed by using MTT, flow cytometry and immunohistochemical staining, respectively. Bone resorption activity of OCs was examined.

Results: The combination of thalidomide (purchased from TTY Biopharm, Taiwan) and liposomal doxorubicin (purchased from TTY Biopharm, Taiwan) profoundly inhibited the amount of harvested viable OCs. The expression of osteoclast-specific surface antigen CD51/61 was markedly inhibited by each drug and their combination. Specifically, the amount of multinucleated TRAP-positive cells characteristics of OCs and bone resorption activity of OCs were suppressed by each drug and, the mostly, their combination.

Conclusion: Thalidomide, liposomal doxorubicin and their combination, beyond currently clinical indications, might be effective regimen to inhibit human osteoclastogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2148. doi:10.1158/1538-7445.AM2011-2148
Abstract 215: Soluble guanylyl cyclase α1 and p53 cytoplasmic sequestration: A novel mechanism for p53 down-regulation in prostate cancer
Authors: Shemshedini Lirim , Gao Shuai
Published: June 2012

Our lab has identified soluble guanylyl cyclase α1 (sGCα1) as a novel androgen- regulated gene involved in prostate cancer cell proliferation. A well-known biological function of sGCα1 is to heterodimerize with sGCβ1 forming the enzyme sGC, which mediates nitric oxide(NO) signaling. Our published data demonstrated that the sGCα1 effect on prostate cancer cell growth is independent of the classical mediators of NO signaling, suggesting that sGCα1 acts via a novel signaling pathway. Furthermore, sGCα1 expression is highly elevated in prostate tumors and this expression is directly correlated to the progression stage of prostate cancer. Interestingly, we have recently discovered that sGCα1 can inhibit the transcriptional activity of p53 via a mechanism that does not depend on classical mediators of NO signaling.

Immunoprecipitation and immunocytochemistry assays have shown an association between sGCα1 and p53 and these data suggest that sGCa1 inhibits p53 by mediating the cytoplasmic sequestration of this tumor suppressor protein, which would represent a previously unknown mechanism for p53 regulation. sGCa1 can also regulate in a gene-specific manner p53-regulated gene expression, targeting genes involved in apoptosis. In addition, sGCa1 makes prostate cancer cells resistant to the chemotherapeutic and apoptosis-inducing drug Etoposide. Finally, analysis of prostate tumors has shown a direct correlation in expression between sGCα1 and p53. Collectively, these data suggest that sGCa1 regulation of p53 activity is important in prostate cancer biology and may represent an import mechanism of p53 down-regulation in those prostate cancers that express significant levels of p53.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 215. doi:10.1158/1538-7445.AM2011-215
Abstract 2181: Tumor suppressor role of Notch3 in medullary thyroid carcinoma
Authors: Jaskula-Sztul Renata , Eide Jacob , Kunnimalaiyaan Muthusamy , Chen Herbert
Published: June 2012

Introduction: NOTCH3 is a transmembrane receptor that appears to be absent in the neuroendocrine (NE) tumors. We have previously demonstrated that transient expression of the active portion of NOTCH3 (NICD3) alters the NE phenotype and inhibits the proliferation of carcinoid tumor cells in vitro. However, the long term effect of activation this pathway is not clearly understood in Medullary Thyroid Carcinoma (MTC). To elucidate the role of Notch3 in NE, we established a doxycycline inducible NOTCH3 intracellular domain in MTC TT cells. The aim of this study was to assess the long term effect of NOTCH3 and determine the mechanism of inhibition of TT cells growth.

Methods: Doxycycline inducible TT-NOTCH3 cells were created by stable transfection of Tet-On vector followed by transfection of pRevTRE-NOTCH3 plasmid. Functional analysis of NICD3 was done by measuring the degree of luciferase activity by CBF binding assay. Quantification of HES and HEY gene expression (Notch signaling mediators) was done by real time RT-PCR. Effect of Notch3 on NE markers -chromogranin A (CgA) and achaete-scute complex-like1 (ASCL1) – was assessed by Western blot analysis. The influence of NOTCH3 overexpression on TT-NOTCH cell proliferation was measured by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) rapid colorimetric assay. The mechanism of growth inhibition was determined for apoptosis markers by Western blot and flow cytometry analyses.

Results: Treatment of TT-NOTCH3 cells with doxycycline led to an induction of NOTCH3 protein in a dose-dependent manner. Increase in the CBF1 binding activity showed that the NOTCH3 protein is functional and this is associated with changes in transcriptional level of HES and HEY families. More importantly, NOTCH3 activation led to a dose dependant reduction of NE markers ASCL1 and CgA. In addition, NOTCH3 activity is required to suppress MTC cell proliferation and the level of growth regression depends on the amount of NOTCH3 protein expressed. Finally, Western blot analysis and flow cytometry experiments indicated that the growth inhibition is due to apoptosis.

Conclusions: We demonstrate, for the first time, that overexpression of NOTCH3 in MTC cells suppresses tumor cells growth by promoting apoptosis. Moreover, Notch3 pathway is functional as active NOTCH3 triggers the CBF-dependent transcriptional network. Finally, the activation of Notch3 in the in vitro model resulted in down-regulation of NE markers indicated that this pathway is conserved in MTC. Therefore, activation of Notch3 could be a therapeutic strategy to treat patients with MTC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2181. doi:10.1158/1538-7445.AM2011-2181
Abstract 2190: ADAMTS8 at 11q25 is a tumor suppressor antagonizing Ras-signaling and methylated in multiple carcinomas
Authors: Choi Gigi C. , Wang Yajun , Li Jisheng , Li Lili , Ying Jianming , Ma Brigette B. , Tao Qian , Chan Anthony T.
Published: June 2012

As opposed to most matrix metalloproteinases which facilitate tumor progression, a growing list of ADAMTS secreted protease members was reported to display tumor suppressive properties with their expressions frequently altered in diverse types of tumors. Here, ADAMTS8 was shown to be located at an LOH locus at 11q25 by array CGH. We further showed that ADAMTS8 was frequently downregulated or silenced in multiple cancer cell lines while broadly expressed in normal tissues. Analysis on the methylation status of the ADAMTS8 promoter CpG island showed that methylation was cancer-specific and ADAMTS8 methylation was correlated with the silencing of ADAMTS8. In addition, expression of ADAMTS8 could be restored with pharmacological or genetic demethylation and hence indicating that CpG methylation directly contributes to the downregulation of ADAMTS8. Aberrant methylation was also frequently detected in primary NPC, gastric and colorectal tumors, but seldom in normal tissues. Ectopic expression of ADAMTS8 led to marked inhibition of cell growth in colony formation assays and cell migration in wound healing assay along with the reduction of phosphorylated ERK protein. The inhibitory effect on tumor cell growth and Ras-ERK pathway by ADAMTS8 indicates its role as a functional tumor suppressor.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2190. doi:10.1158/1538-7445.AM2011-2190
Abstract 2191: The Wnt/DVL signaling antagonist DACT2 is a pro-apoptotic tumor suppressor epigenetically silenced in multiple tumors and inhibits tumor cell proliferation and migration
Authors: Fan Yichao , Ying Ying , Li Lili , Chan Anthony TC , Tao Qian
Published: June 2012

Wnt/DVL/β-catenin signaling pathway is aberrantly activated in a wide range of human cancers which frequently involves the disruption of its negative regulators, hence we would like to identify tumor suppressor as the antagonist to this signaling pathway. We identified a DACT (Dpr/Frodo) family member DACT2 as a downregulated gene in human tumors. Further epigenetic study revealed that, in contrast to its broad expression in normal tissues and cell lines, DACT2 is silenced or downregulated in multiple tumor cell lines including nasopharyngeal (NPC), esophageal (ESCC), gastric, colorectal (CRC), breast, lung, renal (RCC), hepatocellular (HCC), prostate and cervix carcinomas, due to its promoter methylation which could be reversed by pharmacological demethylation. DACT2 promoter methylation was also frequently detected in multiple primary tumors, suggesting that DACT2 is a potential tumor marker. DACT2 encodes a cytoplasm protein localized in endosome. Ectopic expression of DACT2 dramatically inhibited tumor cell colony formation in silenced tumor cell lines, mainly through inducing apoptosis. DACT2 interacts and downregulates Dishevelled (DVL) protein, thus protecting glycogen synthase kinase 3β (GSK-3β) from inactivation by WNT/DVL and allowing GSK-3β to form a complex with Axin and APC to promote the phosphorylation and proteasomal degradation of β-catenin. Overexpression of DACT2 disrupted β-catenin activation and accumulation in nucleus, thus preventing its binding to transcription factors of the Lef/Tcf family. This caused the downregulation of target oncogenes such as c-Myc, CCND1 and fibronectin and the inhibition of tumor cell proliferation and migration. We also observed that DACT2 could upregulate epithelial cell marker E-cadherin, and modulate Akt/GSK-3β/β-catenin signaling pathway, another pathway involved in tumor cell epithelial mesenchymal transition. Our data demonstrate that DACT2 is a functional tumor suppressor epigenetically silenced in multiple tumors which acts as a negative regulator of the Wnt/DVL/β-Catenin pathway, and could be a promising biomarker for human tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2191. doi:10.1158/1538-7445.AM2011-2191
Abstract 2206: SOCS1 and SOCS3 attenuate Lck-mediated cellular transformation through distinct mechanisms
Authors: Venkitachalam Srividya , Chueh Fu-Yu , Yu Chao-Lan
Published: June 2012

The suppressor of cytokine signaling (SOCS) proteins are negative feedback inhibitors of Janus kinase (JAK) – signal transducer and activator of transcription (STAT) signaling in response to cytokine stimulation. Among eight SOCS family members, SOCS1, SOCS2, SOCS3 and CIS (cytokine-inducible SH2-containing protein) are best characterized both structurally and functionally. We showed previously that CIS, SOCS1 and SOCS3 were not expressed in Lck-transformed cells (Cell Signal 18:851; Int J Oncol 36:1201). It suggests that defective negative feedback control may contribute to constitutive JAK-STAT5 activation in Lck-mediated oncogenesis (J Immunol 159:5206; Mol Cancer Res 4:39). These findings also raise the possibility that SOCS proteins may function as tumor suppressors. Indeed, by enforced SOCS1 and SOCS3 expression, we were able to reduce Lck-induced cellular transformation (Int J Oncol 36:1201). In this report, we further examine the molecular mechanisms underlying SOCS tumor-suppressing activity. Through comparative studies of SOCS1, SOCS2, SOCS3 and CIS, we demonstrate here two distinct properties of SOCS1 and SOCS3 toward oncogenic Lck kinase. First, SOCS1 has the highest affinity in binding to oncogenic Lck kinase. We identified the positive regulatory phospho-tyrosine 394 residue (Y394) in the kinase domain as the key interacting determinant in Lck. Additionally, the Lck kinase domain alone is sufficient to bind SOCS1. While the SH2 domain in SOCS1 is important in its association with the oncogenic Lck kinase, other functional domains may also contribute to overall binding affinity. These data suggest that SOCS1 may inhibit Lck kinase activity in a manner similar to JAK kinases. Second, only SOCS3 becomes phosphorylated in the presence of oncogenic Lck kinase. We identified the phosphorylation sites as Y204 and Y221 located in the C-terminal SOCS box of SOCS3. We further analyzed the contribution of SOCS3 tyrosine phosphorylation on its tumor suppressor activity. Preliminary studies suggested that SOCS3 phosphorylation on Y204 and Y221 might play an important role in blocking cell proliferation and cell cycle progression of Lck-transformed cells. SOCS3 tyrosine phosphorylation can lead to altered protein stability and crosstalk with other signal transduction pathways, such as MAPK signaling. Experiments are in progress to further define the molecular details of SOCS3 phosphorylation involved in tumor suppression. All together, these results provide important mechanistic insights into the distinct roles of SOCS proteins as tumor suppressors in cells transformed by oncogenic protein tyrosine kinases. This work is supported in part by NIH grant CA107210.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2206. doi:10.1158/1538-7445.AM2011-2206
Abstract 2208: Overexpression of NDRG1 inhibits cell migration of breast cancer cells via reactive oxygen species during reoxygenation
Authors: Lai Liang-Chuan , Su Yi-Yu , Chen Kuo-Chih , Tsai Mong-Hsun , Sher Yuh-Pyng , Lu Tzu-Pin , Chuang Eric Y.
Published: June 2012

One characteristic of tumor microenvironment is oxygen fluctuation, which results from hyper-proliferation and abnormal metabolism of tumor cells as well as disorganized neo-vasculature. Reoxygenation in tumor microenvironment can induce oxidative stress, which further leads to DNA damage and genomic instability. Although multiple cellular responses are activated in order to survive under this microenvironment, little is known about the dynamic response upon reoxygenation. Therefore, in order to investigate the transcriptional response of tumor adaptation to reoxygenation, breast cancer cells MCF-7 was cultured under 0.5% of hypoxia for 24h followed by 24h of reoxygenation in normoxia. Cells were harvested respectively at 0, 1, 4, 8, 12, and 24h during reoxygenation. The genomic profiling of MCF-7 upon reoxygenation was examined using Illumina Human-6 v3 BeadChips. We identified 127 differentially expressed genes in which 53.1% were up-regulated and 46.9% were down-regulated upon reoxygenation. Pathway analysis revealed that HIF-1-alpha transcription factor network was most significantly (P=2.02×10-3) enriched in these differentially expressed genes. Among these genes, a group of selected genes of interest were further validated by quantitative real-time PCR. In particular, NDRG1 was highly suppressed upon reoxygenation. NDRG1 (human N-myc down-regulated gene 1) is associated with a variety of stress and cell growth-regulatory conditions. To characterize whether NDRG1 play a role in reoxygenation, NDRG1 protein was overexpressed in MCF-7 cells. Upon reoxygenation, overexpression of NDRG1 inhibited cell migration, and this inhibition could be reversed by adding reactive oxygen species scavengers. Our results revealed dynamic expression profiling of MCF-7 upon reoxygenation, and demonstrated that NDRG1 is involved in tumor adaptation to reoxygenation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2208. doi:10.1158/1538-7445.AM2011-2208
Abstract 222: Differential p73 isoform expression profile in non-Hodgkin's lymphoma with 1p36 chromosomal abnormality modulates angiogenic phenotype
Authors: Hassan Hesham M. , Varney Michelle L. , Jain Smrati , Joshi Shantaram S. , Singh Rakesh K. , Dave Bhavana J.
Published: June 2012

Alteration of tumor suppressor gene function often leads to imbalance in expression of angiogenic and angiostatic factors facilitating disease progression. TP73, a member of the p53 tumor suppressor gene family, regulates many important cellular pathways and has been shown to be abnormally expressed in non-Hodgkin's lymphoma (NHL). TP73 gene function is controlled by harmony between two isoforms with opposing functions, the full length TAp73 protein (pro-apoptotic) and the NH2-terminally truncated ΔNp73 protein (anti-apoptotic through antagonizing both TAp73 and p53). We have observed differential ΔNp73 up-regulation and increase in ΔNp73:TAp73 expression ratio among NHL cases with 1p36 chromosomal disruption as compared to NHL cases without 1p36 disruption, which is associated with aggressiveness. To decipher whether deregulated p73 isoforms expression modulates the angiogenic potential in NHL we examined the mRNA expression of angiogenic factors (VEGF-A, FGF-2, and angiogenic chemokine CXCL-8) and angiostatic factors (TSP-1 and Endostatin) in NHL cases with and without chromosome 1p36 abnormality by real-time PCR. We observed higher levels of expression of angiogenic factors (VEGF-A, FGF-2, and CXCL-8) and lower levels of angiostatic factors (TSP-1 and Endostatin) in NHL cases with 1p36 abnormality as compared to NHL cases without 1p36 abnormality. Next, we performed a non-parametric bivariate correlation analysis (Kendall's tau_b and Spearman's rho) to examine whether ΔNp73:TAp73 expression ratio correlates with the expression of angiogenic and angiostatic factors. We observed a significant correlation between ΔNp73:TAp73 expression ratio with the expression of VEGF-A (r=0.900, p<0.001), FGF-2 (r=0.620, p<0.05) and TSP-1 (r=−0.873, p<0.001). To further elucidate the relationship between differential p73 isoforms expression and the expression pattern of angiogenic and angiostatic factors, we stably transfected an NHL cell line with 1p36 disruption, Granta-519 (TAp73 non expressor) with a wild type TAp73 mammalian expression vector. We observed an up-regulation of TSP-1 and Endostatin expression among Granta-519 cells transfected with TAp73 vectors compared to isogenic control vector transfected cells. Our data highlights the molecular and functional consequences of the differential p73 isoforms expression pattern caused by 1p36 chromosomal disruption on angiogenic phenotype in NHL.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 222. doi:10.1158/1538-7445.AM2011-222
Abstract 223: ΔNp63α, a p53 family member protein, promotes tumor progression in pancreatic cancer
Authors: Danilov Alexey , Neupane Divas , DiRenzo James , Korc Murray
Published: June 2012

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths in both men and women. Targeted therapies thus far have failed to demonstrate survival advantage in PDAC and novel therapies are needed. Mutations in Ras and p53 are frequently found in PDAC and promote cancer cell survival, invasiveness and chemoresistance. In contrast, p63, an ancestral member of p53 family, is rarely mutated in cancer. ΔNp63α was shown to be overexpressed in squamous cell tumors of the lung and head and neck and thus has been implicated in carcinogenesis. We tested expression of p63 isoforms in five PDAC cell lines and investigated its role in tumor progression.

Expression of p63 was determined by immunoblotting, real-time quantitative and semi-quantitative PCR in 5 PDAC cell lines. PANC-1 cells, with low levels of p63, were transfected with ΔNp63α cDNA-expressing vector. Conversely, high endogenous p63 levels in T3M4 and BxPC3 cell lines were suppressed by specific shRNA. We studied cell proliferation in a doubling assay, motility in wound assay and invasiveness in Matrigel chambers.

We found that pancreatic cancer cell lines predominantly expressed ΔN isoform of p63, and specifically ΔNp63α variant. ΔNp63α protein and transcript levels were high in T3M4, BxPC3 and Colo-357 cell lines and low in ASPC-1 and PANC-1 cell lines. Interestingly, transcript levels of TAp63, a p63 variant implicated in cellular senescence and suppression of metastases, were near the detection threshold in all cell lines. Overexpression of ΔNp63α in PANC-1 cells resulted in increased proliferation and enhanced migration of those cells but had no effect on invasiveness in serum-free conditions. Incubation of ΔNp63α-expressing PANC-1 cells in presence of epidermal growth factor increased both migration and invasiveness. By contrast, shRNA knockdown of endogenous p63 in BxPC3 cells led to decreased proliferation. Silencing p63 in T3M4 cells resulted in attenuated migration in serum-free conditions and decreased migration and invasiveness in response to epidermal growth factor.

Thus, ΔNp63α is a predominantly expressed isoform of p63 in PDAC cell lines. ΔNp63α plays an important role in PDAC progression and thus its associated pathways may potentially be therapeutically targeted in this disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 223. doi:10.1158/1538-7445.AM2011-223
Abstract 2265: Clinical significance of HER3 expression in curatively resected pancreatic cancer
Authors: Hirakawa Toshiki , Nakata Bunzo , Kashiwagi Shinichiro , Kato Yukihiro , Fuyuhiro Yuhiko , Iwauchi Takehiko , Matsusoka Junko , Itoh Gentaro , Aomatsu Naoki , Yoshii Mami , Hasegawa Tsuyoshi , Amano Ryosuke , Kubo Naoshi , Tanaka Hiroaki , Muguruma Kazuya , Yamada Nobuya , Yashiro Masakazu , Sawada Tetsuji , Ohira Masaichi , Hirakawa Kosei
Published: June 2012

Introduction: HER3 (erbB-3) is a member of the epidermal growth factor receptor (EGFR) family. After dimerization with other members of the EGFR family several signal transduction cascades can be activated.HER2/HER3 heterodimer is likely to be the most effective complex for activating downstream pathways.Because of special importance of HER2/HER3 heterodimer in tumorgenesis, the signaling pathways and downstream effectors of the HER family become key molecules in the strategy of cancer therapy.HER3 is overexpressed in diverse human cancers, and has been associated with poor prognosis in breast, lung, and ovarian cancer.The objective of this study was to evaluate the significance of HER3 expression in pancreatic cancer cells.

Material and Methods: A total 86 patients were enrolled.The pancreatic cancers were stained with antibodies against HER3 by immunohistochemistry. HER3 expression was evaluated by intensity of staining.Intensity was given scores 0-3 (score 0:no staining, score1+:barely perceptible staining, score2+:moderate staining, score3+:strong staining). Scores of 0 and 1+ were considered to be negative for HER3 expression, while 2+ and 3+ were considered to be positive (overexpression).We examined the association between HER3 expression and other clinicopathologic variables.

Results: HER3 expression was positive in 36 (41.9%) cases.There were no significant correlation between HER3 expression and other clinicopathological factors. The prognosis of HER3 positive patients was significantly poorer than that of HER3-negative patients (p=0.002). A univariate survival analysis revealed that HER3 overexpression, intrapancreatic nerve invasion and T category were significantly correlated with patient survival. A multivariate analysis showed HER3 overexpression was an independent predictor of worse prognosis in resectable pancreatic cancer.

Conclusion: The expression of HER3 might be a novel predictive prognostic marker for patients with pancreatic cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2265. doi:10.1158/1538-7445.AM2011-2265
Abstract 2273: Validation of putative stem cell factor SOX2 as predictive marker in breast cancer – with overall poor prognosis
Authors: Pradel Leon Paul , Swierczynski Stefan , Greil Richard , Dietze Otto , Hauser-Kronberger Cornelia
Published: June 2012

As member of the high-mobility-group (HMG)-domain containing proteins – SOX2 (Sex determining Region Y – box 2) transcriptional activator is known for its pivotal role in maintaining self-renewal, pluripotency and cellular identity in embryonic stem cells. While its status in adult tissue, e.g. in homeostasis – is still less understood and controversy – SOX2 has shown recently to be up regulated in a huge variety of human tumors like oesophagus, lung, brain and breast.

Within this study whole sections – provided as formalin-fixed paraffin-embedded tumor samples, derived from 86 patients with advanced breast cancer disease were stained via immunohistochemistry (IHC) for SOX2 positivity. Further on – six breast cancer cell lines (MCF-7, MDA-MB231, MDA-MB453, SK-BR3, SUM-159, T47-D) as well as an individual based tissue micro array (TMA) containing 1080 fine needle biopsies were additionally analyzed using IHC and qRT-PCR. Initially – all samples have been evaluated for their particular molecular subtype (Luminal A, Luminal B, Normal-like, HER2 Overexpressing, Basal-like) and their histopathological parameters (histological subtype, grade, size, lymph node metastasis).

For further proof of principle the applied SOX2 polyclonal antibody (SOX2 PAb AF2018, R&D SystemsTM) was validated for its marker capacity via Surface Plasmon Resonance (SPR) Spectroscopy and Western Blot to determine the kinetic profile, limit of detection, epitope variety and specificity of this antibody. With the so called ‘fishing’ opportunity for analyte recovery in SPR, native putative antigen bound by the SOX2 PAb can be enriched and further characterized from whole cell lysates.

In conclusion – SOX2 showed association to overall bad prognosis due to high grade (p<0.001), huge size (p=0.003), lymph node metastasis (p<0.001) as well as histological and molecular subtypes with poor overall-survival (p<0.001). Further on it is independent (p=0.6) of oestrogen receptor (ER) presence but correlated to EGFR expression (p=0.007). During antibody-based validation – SOX2 PAb revealed to possess high specificity, sensitivity and affinity to native and recombinant SOX2 full length protein finally. In this way SOX2 – as marker, can be applied to predict overall poor prognosis in breast cancer in an ER- and subtype-independent fashion.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2273. doi:10.1158/1538-7445.AM2011-2273
Abstract 2276: Serum endoglin and uPA change predicts PFS and overall survival in first line trastuzumab-treated breast cancer
Authors: Evans Matthew S. , Hou Hui Ying , Anyanwu Uchechi , Leitzel Kim , Ali Suhail M. , Koestler Wolfgang , Fuchs Eva-Maria , Brown-Shimer Sheryl , Hamer Peter , Carney Walter , Lipton Allan
Published: June 2012

Background: About half of HER2-positive breast cancer patients will respond to first-line trastuzumab-containing therapy, but of these most will progress within a year with acquired resistance. Since trastuzumab treatment is now used in the HER2-positive adjuvant breast cancer setting, trastuzumab resistance will continue to be a recurring clinical problem, and better predictive and prognostic biomarkers are urgently needed.

Methods: Serum TIMP-1, uPA, carbonic anhydrase 9 (CA9), and endoglin (co-receptor for TGF- β) were measured using ELISA in 60 metastatic breast cancer patients before starting first-line trastuzumab-containing therapy, and at the closest 1 month post-trastuzumab blood draw. The TIMP-1, uPA and CA9 ELISAs were from Oncogene Science/Siemens Healthcare Diagnostics, and the endoglin ELISA was from R&D Systems. Progression-free (PFS) and overall survival (OS) were analyzed using the Kaplan-Meier method and Cox modeling.

Results: When serum biomarker change (pretreatment to 1 month post-trastuzumab treatment) was analyzed on a continuous basis, change in serum TIMP-1 and CA 9 were not significant for PFS or OS. However the change in serum endoglin (HR = 1.017, p = 0.013) and serum uPA (HR 1.018, p = 0.006) were significant predictive factors for PFS to first-line trastuzumab treatment. The change in serum endoglin (HR = 1.021, p = 0.004) and serum uPA (HR 1.016, p = 0.03) were also significant prognostic factors for OS: declining serum endoglin or uPA predicted for increased PFS and OS. Changes in serum endoglin and uPA were positively correlated (r = 0.7, p <0.0001). Due to the significant correlation between the change in serum endoglin and uPA levels, these 2 serum biomarkers were analyzed separately along with the change in serum TIMP-1 and CA9 in multivariate analysis. In these analyses, only the change in serum endoglin or uPA was significant.

Conclusions: The change in serum endoglin and uPA level from pretreatment to 1 month post-trastuzumab treatment predicted PFS and overall survival in metastatic breast cancer patients treated with first-line trastuzumab-containing therapy. Endoglin (CD105) is a co-receptor for TGF-β, is expressed by human vascular endothelial cells, and plays a major role in angiogenesis. These angiogenesis-related serum biomarkers deserve further study in larger trials of HER2-targeted breast cancer treatment.

Supported by a grant from Komen for the Cure.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2276. doi:10.1158/1538-7445.AM2011-2276