bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3872: Differentially expressed stem cell markers in breast cancer stem cells
Authors: Lobba Aline RM , Forni Maria Fernanda PAD , Perozzi Carolina , Carreira Ana Claúdia O. , Labriola Leticia , Sogayar Mari Cleide
Published: June 2012

The recently emerged concept of cancer stem cell (CSC) has led to a new hypothesis about tumor progression. Basically, the CSC theory hypothesizes the presence of a hierarchically organized, relatively rare population of cells, which is responsible for tumor initiation, self-renewal and maintenance, in addition to mutation accumulation and resistance to chemotherapy. CSCs have recently been described in breast cancer. Different markers have been used to isolate breast cancer stem cells; however, none of these proved to be very efficient. Furthermore, the lack of specific markers limits a precise characterization of these cells and their clinical application, especially in the development of therapeutic protocols. In the present work, we set out to search for enriched subpopulations of CSCs. To this end, we investigated the profile of different surface markers in normal and tumoral breast cancer cell lines. We analyzed the expression of cd44, cd31, cd90, cd200, cd14, cd34, cd31, cd105, c-kit, abcg2 and cd29 in MCF10-A, MCF-7, MDA-MB-231, MDA-MB-435 and Hs578-T cells lines using quantitative RT-PCR. As a result of this effort, we were able to identify two distinct markers presenting differential expression between normal and tumor cells. While one marker was increased in the malignant cells lines, the other one presented higher levels of expression in the normal cell line. Based on this study, we propose a new set of markers, which should allow both isolation and characterization of breast cancer stem cells. Upon informed consent, a number of normal and tumor human breast samples are currently being collected for the final proof of concept. These results contribute not only to a deeper understanding of the Biology of CSCs, but, also, to the development of optimized therapeutical approaches to breast cancer.

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 3872. doi:10.1158/1538-7445.AM2011-3872
Abstract 3874: An isogenic cell line approach to MMTV integration site analysis
Authors: Knepper Janice E. , Acosta Lauren , Saunders Justin , Tio Chong-Wai , Wang Tianjiao , Danielson Keith G.
Published: June 2012

Development of mammary tumors in mice carrying the retrovirus, mouse mammary tumor virus (MMTV), occurs by insertional mutagenesis in which viral transcriptional regulatory elements drive increased expression of adjacent genes. Several oncogenes have been identified due to their ability to cause loss of cell growth control in this system. However, most virus positive tumors feature multiple integration sites, and determination of which may be significant in tumor growth represents a challenge to validation of importance of integration site genes. We have developed a panel of more than 20 independent cell lines derived from MMTV-induced tumors of BALB/c mice. Analysis of common integration sites led to the identification of Tmem170, encoding a predicted transmembrane protein of 144 amino acids. Tmem170 RNA is highly expressed in several cell lines in the panel, but not in normal mouse mammary gland. Rabbit antiserum against a peptide in the predicted extracellular domain revealed a single band migrating at approximately 40 KDa in SDS gels. We developed a panel of vectors encoding silencing RNAs targeting Tmem170 and introduced these into cell lines expressing the Tmem170 RNA. Two independent cell lines exhibited decreased doubling times in culture when Tmem expression was silenced. The cells were introduced into the mammary fat pads of isogenic BALB/c mice and silenced lines exhibited dramatically decreased tumor growth. These studies support Tmem170's oncogenic role in tumor growth. In addition, the use of an easily manipulated cell line model for assessing the contribution of upregulated genes to tumorigenesis in immune competent animals allows for testing of other potential oncogenes.

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 3874. doi:10.1158/1538-7445.AM2011-3874
Abstract 3888: Ionizing radiation destabilizes inverted ALU repeats linked to LOH
Authors: Ho Bay , Singh Sheetal , Shih Shyh-Jen , Baker Pamela M. , Do To Uyen , Vaughan Andrew T.M
Published: June 2012

Ionizing radiation is a potent inducer of Loss of Heterozygosity (LOH), driven primarily by DNA double strand breaks. Such a process impacts both tumorigenesis through deletion of linked tumor suppressor genes and, similarly, the potential development of radiation resistant tumor clones during radiation therapy. The human mucoepidermoid cell line, H292, was exposed to either 4 or 8 Gy and surviving clones developed from two rounds of cloning analyzed for LOH using polymorphic microsatellite markers. A high level of LOH was observed within individual clones extending from 11q21 to 11q24 (6/49 after 4 Gy and 8/50 after 8Gy). More detailed analysis using a SNP screen on selected LOH positive clones indicated a sharp telomeric termination of LOH adjacent to a common SNP location. The telomeric extent of LOH observed co-mapped to a region of elevated recombination instability documented previously using linkage analysis. Further, SNP analysis showed a region of approximately 1 Mbp LOH that included the region of recombination instability identified through linkage analysis, in some clones isolated either with or without irradiation. The consistency of the telomeric boundary of LOH observed in the irradiated clones permitted a targeted analysis of DNA double strand breaks at this location, using LM-PCR, both before and after 4Gy irradiation of the same cell line. Using a six primer set screen covering a 9 kbp region at the telomeric extent of radiation-induced LOH, a single primer set showed an elevation in DNA breaks from 2-8h after irradiation. The timing and discrete distribution of such breaks indicated that these were not directly induced by the radiation itself. Cells that were unirradiated were not affected. Further inspection of the location demonstrating selective radiation-linked fragmentation showed the presence of an ALU inverted repeat, separated by approximately 1kbp, where the DNA breaks determined by LM-PCR were clustered within the 5’ ALU. Others have shown previously that inverted ALU repeats separated by <20bp are recombinationally unstable and are underrepresented within the human genome. It is proposed that, within the H292 cell line, otherwise stable ALU inverted repeats are capable of forming a transient hairpin conformation, perhaps facilitated by their own transcription via RNA polymerase III, which is unstable within a genome undergoing DNA damage and repair. The combination of such inverted ALU repeats and genotoxic exposure, such as during radiation therapy, may therefore destabilize specific regions of the genome and contribute to the development of aberrant cell clones.

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 3888. doi:10.1158/1538-7445.AM2011-3888
Abstract 390: Increased orthotopic pancreatic tumor cell growth in obese mice
Authors: Gorden Lee , VanSaun Michael N.
Published: June 2012

Pancreatic cancer remains one of the eight deadliest cancers due to its late detection and high propensity to metastasize. Increased risk for the development of pancreatic cancer has been associated with obesity, type II diabetes and pancreatitis. The contribution of adipocytes and adipokines to pancreatic cancer progression remains largely unknown and requires further analysis. We hypothesize that increased pancreatic fat compromises the pancreatic microenvironment and increases primary tumor growth as well as metastasis. The condition of non-alcoholic fatty pancreatic disease has been characterized in humans as well as mice and lymph node positive patients were found to have increased pancreatic fat1,2. Adipose secreted cytokines, also known as adipokines, have both been implicated in various aspects of cancer cell survival, growth, and migration. To understand the influence that increased pancreatic fat has on the growth of tumors, both primary and metastatic, we have investigated a high fat diet induced murine model. Male mice fed a 42% fat diet for three months develop increased inter-pancreatic and intra-pancreatic fat as well as acute localized pancreatitis when compared to mice fed a 13% fat diet. To test the affect of pancreatic fat on tumor cells, we used an orthotopic tumor injection model to determine the rate of primary tumor growth and progression to metastasis. Further, we are implementing the use of Panc02-luciferase tagged cells to monitor and measure pancreatic tumor growth over time and to detect the presence of metastases in the liver and/or lungs. Preliminary evidence has confirmed that orthotopic tumors grown in obese mice led to an increased tumor size as well as massive expansion of tumor cells into the interpancreatic fat. In conclusion, increased pancreatic fat promotes the growth of orthotopically implanted murine tumor cells, yet the molecular mechanisms remain undetermined.

1. Mathur, A. et al. J Am Coll Surg. 2009 May;208(5):989-94.

2. Mathur, A. et al. HPB (Oxford). 2007;9(4):312-8.

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 390. doi:10.1158/1538-7445.AM2011-390
Abstract 4002: Epigenetic control of a checkpoint for miRNA tolerance
Authors: Li Qunfang , Tainsky Michael A.
Published: June 2012

The interferon (IFN) pathway controls the cellular response to viral infection and double-stranded RNA. We have found that this pathway provides a novel checkpoint for enhanced expression of miRNAs. The IFN pathway is abrogated in fibroblasts from Li-Fraumeni Syndrome (LFS) patients during spontaneous cellular immortalization, a necessary step in carcinogenesis. Interferon stimulated genes (ISGs) were down-regulated by epigenetic silencing during immortalization and some of these same ISGs were activated by replicative senescence. Bisulfite sequencing of the promoter regions of IFN-regulatory transcription factors (IRF5 and IRF7) revealed that IRF7 but not IRF5 was epigenetically silenced by methylation of CpG islands in immortal LFS cells. The re-activation of ISGs by those transcription factors is sufficient to inhibit cell proliferation and induce a senescence-related phenotype in immortal LFS cells. Similar epigenetic silencing of IRF7 also results in disruption of IFN pathway in ∼30% lung cancer cells and primary tissues. Examination of innate immunity status in normal bronchial epithelial cell line Beas2B, its ras transformed derivative BVK cells and tumor-generating clones reveals increasing loss of functional IFN pathway as cells become more cancerous. Furthermore, microarray profiling of differentially expressed microRNAs (miRNAs) in LFS cell lines revealed most miRNAs were up-regulated in IFN-pathway-defective MDAH087-10 fibroblasts compared to MDAH087-N cells with relatively normal IFN signaling. Overexpression of Dicer, a critical enzyme in miRNA biogenesis, promoted cell growth and colony formation in MDAH087-10 cells. However, double-stranded miRNA produced by Dicer enhanced the expression of ISGs in MDAH087-N cells resulting in significant cell death and reduced cell growth. In addition, manipulation of the IFN pathway in immortal LFS fibroblasts through transcription factor IRF7 reversed their response to Dicer overexpression due to altered activation of the IFN pathway. Dicer-overexpressing MDAH087-N cells contained lower levels of miRNA than vector control, and conversely much higher miRNA expression was detected in Dicer transfected MDAH087-10 cells. The higher miRNA tolerance in immortal LFS fibroblasts with a disrupted IFN pathway indicates for the first time that the IFN pathway as mediated through the transcription factor IRF7 must be disrupted to permit miRNA up-regulation to occur in early carcinogenesis. This IFN pathway appears to provide a checkpoint for miRNA level tolerance and its abrogation by epigenetic silencing leads to cellular immortalization and early events in tumorigenesis.

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 4002. doi:10.1158/1538-7445.AM2011-4002
Abstract 4004: Mislocalization of ATBF1 is associated with progression of head and neck squamous cell carcinoma
Authors: Sun Xiaodong , Li Jie , Sica Gabriel , Fan Song-Qing , Wang Yuxiang , Chen Zhengjia , Muller Susan , Chen Zhuo G. , Shin Dong M. , Dong Jin-Tang
Published: June 2012

Purpose: In prostate, breast and gastric cancers, the nuclear protein AT-motif Binding Factor 1 (ATBF1) undergoes frequent reduced mRNA expression and genomic alterations. However, the protein level and subcellular localization of ATBF1 have not been well studied in human cancers. In the present study, we examined ATBF1 expression, localization and function in head and neck squamous cell carcinomas (HNSCCs).

Experimental Design: ATBF1 expression and localization were examined by immunohistochemistry in a series of 197 surgically dissected HNSCC specimens and correlated with clinical outcomes. In addition, ATBF1 expression was characterized in five HNSCC cell lines. We then mutated the nuclear localization signal (NLS) of ATBF1 and studied the effects of cytoplasmic versus nuclear ATBF1 on the growth kinetics of the 212LN cell line.

Results: ATBF1 had a predominantly nuclear localization in hyperplastic squamous epithelium, and nuclear ATBF1 dramatically decreased in invasive tumors. Conversely, cytoplasmic ATBF1 levels progressively increased from dysplasia to invasive tumors. Cytoplasmic ATBF1 levels were significantly inversely correlated with overall survival and disease free survival. Similar expression patterns and subcellular localization of ATBF1 were observed in HNSCC cell lines. In order to better define the role of subcellular localization of ATBF1, we identified and mutated its nuclear localization signal (NLS). Mutation of the NLS converted the inhibitory effect of ATBF1 on cell growth to growth-promoting.

Conclusions: Aberrant cytoplasmic localization of ATBF1 is significantly associated progression of HNSCC, and cytoplasmic ATBF1 may be a potential biomarker for its early detection. Our results suggest that nuclear ATBF1 functions as a tumor suppressor in head and neck squamous epithelial cells and that this tumor suppressor effect is sequestered by cytoplasmic localization during HNSCC progression.

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 4004. doi:10.1158/1538-7445.AM2011-4004
Abstract 4239: Pterostilbene, a natural polyphenol, elicits full protection against ultraviolet B radiation-induced skin carcinogenesis: Preclinical studies
Authors: Feddi Fatima , Mena Salvador , Ortega Angel , Asensi Miguel , Estrela Jose M.
Published: June 2012

Solar radiation exposure is the chief cause of nonmelanoma (i e, basal cell and squamous cell) skin cancer, and it is also a prime factor in the etiology of cutaneous melanoma The cancer-causing effects of ultraviolet (UV) radiation on the skin are mainly produced by UV-B radiation in the 290- to 320-nm range, the same range that produces burning in human skin (erythema) UV-B exposure can damage DNA and be immunosuppressive Thus, and considering the alarming numbers of skin cancers being diagnosed around the world, it is increasingly evident the need of effective protection from UV radiation

Resveratrol (trans-3,5,4′-trihydroxystilbene; RES) is a phytoalexin present in a wide variety of plant species, where its synthesis is induced by stress conditions The cancer chemopreventive activity of RES was first reported by Jang et al [Science 275, 218 – 220 (1997)] in a model of skin carcinogenesis where topic administration of this polyphenol inhibited multistage mouse skin carcinogenesis Equally promising action is exerted by resveratrol analogues, mainly pterostilbene (3,5-dimethoxy-4′-hydroxy-trans-stilbene: PTER), which shows a higher half-life in vivo and more potent anticancer effects in vivo than RES Our aim was to compare the anticancer effects of RES and PTER in UVB-induced mouse skin carcinogenesis In our experiments, Female SKH-1 hairless mice were irradiated with UVB (180 mJ/cm2; 3 doses/week; for a total of 30 weeks) 20 min after or before the topical administration of liposomes containing a total of 10 μmol of RES or PTER/mice (aprox 2 5 μmol/cm2 of skin) This long-term UVB administration reproduces, in an animal model, which are the consequences in humans of receiving chronic UVB radiations We observed that pretreatment of the skin with PTER resulted in a dramatic reduction in UVB-induced skin tumorigenesis (90 % of tumor free-mice at the end of treatment, n=20; P<0 01), whereas all RES-treated mice showed carcinomas (0 % of tumor free-mice on the 30th week, n=20) Molecular mechanisms involved in this strong anticancer effect elicited by Pter (as compared with RES) suggest differences in prevention of DNA mutagenesis, and protein and lipid oxidation Our results indicate that PTER can be used efficiently in the prevention of skin carcinogenesis induced by solar radiation.

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 4239. doi:10.1158/1538-7445.AM2011-4239
Abstract 4241: Flavokawain B, a kava chalcone, induces apoptosis in synovial sarcoma cell lines
Authors: Sakai Toshinori , Guo Yi , Ramez Eskander , Mefford Jason , Hopkins Justin , Zi Xiaolin , Bhatia Nitin N. , Hoang Bang H.
Published: June 2012

Synovial sarcoma (SS) is a soft tissue sarcoma with poor prognosis and lack of response to conventional cytotoxic chemotherapy. The regulatory mechanisms for the rapid proliferation of synovial sarcoma poorly understood. Surgical resection with/without adjuvant radiotherapy and chemotherapy are the mainstay of treatment. Although the sarcoma cell lines have moderate chemosensitivity to isofamide and doxorubicin therapy, the overall prognosis is still poor. Therefore, further development of the treatment for SS is required. Kava is an ancient crop of the western Pacific. The root extract of kava has been served as a traditional beverage for Pacific Islanders. Consumption of this traditional beverage has been thought to be correlated with low and uncustomary sex ratios of cancer incidence in 3 kava-drinking countrites: Fiji, Vanuatu and Western Samoa. We have already reported that a kind of fravokawains from kava extracts (FKA: flavokawein A) induced apoptosis against bladder cancer cells, and flavokawein B (FKB) was also significant effective in inhibiting the growth of prostate cancer cells. In this study, we examined the inhibitory proliferation effects of the FKB in synovial sarcoma growth and whether FKB modulates tumor apoptosis cascades in human synovial sarcoma cell lines. We treated 2 cell lines of synovial sarcoma (SYO-I, HS-SY-II) by different concentrations of FKB (0, 2.5, 5.0, 7.5 μg/μl) for 24 hours. From the results of real time PCR, depending on the concentrations of FKB in both cell lines, the expression of Survivin was found to be decreased, while those of DR5, Puma, and Bim were increased. These results indicated that FKB effectively induces apoptosis of the two cell lines depending on the concentration. In conclusion, our findings demonstrate that FKB is effective for growth inhibition of synovial sarcoma cell lines in vitro and suggest that FKB may be a new therapeutic option for patients with synovial sarcoma.

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 4241. doi:10.1158/1538-7445.AM2011-4241
Abstract 4246: Enhanced anti-vascular and anti-tumor effects upon combination therapy with aflibercept (VEGF Trap) and other anti-angiogenic drugs
Authors: Adler Alexander P. , Vangala Pranitha , Castanaro Carla , Daly Christopher , Thurston Gavin , Eichten Alexandra
Published: June 2012

Experimental and clinical development of anti-angiogenic drugs to date has mostly focused on the VEGF pathway, although other signaling pathways, including Dll4-Notch and Angiopoietin-Tie, are now also being explored. Targeting one pathway, however, might not result in maximum anti-vascular and anti-tumor effects. In this study, we compared the rapid effects on tumor perfusion of single agent blockade of VEGF, Dll4 or Ang2 to the effects of combined therapy (VEGF blockade plus either Dll4 or Ang2 blockade), and compared long-term tumor growth responses to these early effects on tumor perfusion. Using aflibercept (VEGF Trap) alone and in combination with a blocking Dll4 antibody or a blocking Ang2 antibody, we compared various tumors with a wide range of long-term growth responses to each agent: C6 rat glioma tumors, HT1080 human fibrosarcoma and Colo205 human colon carcinoma tumors, all grown subcutaneously in SCID/CB17 mice. Tumor vascular perfusion was assessed using contrast-enhanced micro-ultrasound at 24 and 72 hr after treatment. We found that aflibercept treatment rapidly decreased vascular perfusion in C6 (59% after 24 hr) and Colo205 (32%) tumors, whereas there was no decrease observed in HT1080 tumors. These changes in perfusion correlated with long-term tumor growth inhibition upon aflibercept treatment. Dll4 blockade resulted in decreased vascular perfusion in C6 (30% after 24 hr), Colo205 (77%), and HT1080 (54%) tumors, which again correlated with long-term growth inhibition of the different tumors. Treatment with an anti-Ang2 antibody produced a slight decrease in perfusion of Colo205 tumors (13% after 24 hr), associated with a decrease in tumor growth. However, combination treatments with aflibercept plus either anti-Dll4 or anti-Ang2 antibodies were more potent in all tumor types compared to single agent treatments at reducing tumor perfusion as well as inhibiting long-term tumor growth. Taken together, these data show that changes in tumor perfusion correlate with long-term tumor responses to combination treatment of aflibercept plus either anti-Dll4 or anti-Ang2 antibody suggesting that decreases in tumor perfusion could be an early biomarker predictive of response to anti-angiogenic therapies. Measuring the increased effectiveness of combination therapy in patients treated with different anti-angiogenic drugs may facilitate the improvement of current treatment regimens.

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 4246. doi:10.1158/1538-7445.AM2011-4246
Abstract 4247: Iron controlled treatment can be novel therapeutic agent and biomarker of Bevacizumab
Authors: Ohara Toshiaki , Noma Kazuhiro , Watanabe Shinichiro , Ohnishi Teppei , Nishitani Seishi , Tomono Yasuko , Hashimoto Yuuri , Tazawa Hiroshi , Fujiwara Toshiyoshi
Published: June 2012

[Introduction] Lung cancer is one of the most aggressive malignancies with poor prognosis. Many kinds of new medicine for chemotherapy have been developed and introduced in clinical practice, whereas the mortality has not been satisfied. Furthermore, development of new medicine for chemotherapy costs a vast sum of money, which leads expensive treatment. Therefore, It's expected that development of new strategy extremely enhances the efficacy of ordinary therapeutic strategy. Iron metabolism and its relationship with cancer cell have been studied for a long time. Iron overload is known to induce some kinds of cancer, which suggests that prevention of iron overload could be a therapeutic strategy of cancer prevention. In fact, iron deficiency is also known to suppress the tumor growth in vivo study. However the efficacy is not superior to the ordinary chemotherapy, it's not a standard therapeutic strategy of cancer. In this study, we demonstrated that iron deficiency induced the suppression of the tumor growth and unwillingly up-regulation of angiogenesis in vitro and in vivo studies. Moreover using Bevacizumab together strongly suppressed the tumor growth in NSCLC. This result suggests that Iron controlled treatment can be novel therapeutic agent and biomarker of Bevacizumab.

[Materials and Methods] A549, H1299 NSCLC cell lines were used. In vivo study, Normal diet and Iron deficient diet are prepared. Normal diet and Iron deficient diet were fed for 3 weeks before implantation of A549 cells in athymic male nude mice(n=8/group). In Bevacizumab administration study, as shown above, Bevaicuzmab(5 mg/kg) was administrated twice a week in each diet group. In vitro study, Iron chelator deferasirox was used in vitro study. Cell viability was determined by WST-1 assay. Cell cycle analysis was measured by flow cytometry. To determine angiogenesis via hypoxia, western blotting analysis and VEGF ELISA assay were done.

[Results] Tumor growth was suppressed in iron deficient diet group (p=0.0037). Bevacizumab revealed to have an additive effect to inhibit the tumor growth. Deferasirox reduces A549 and H1299 cell proliferation in a concentration manner via cell cycle arrest. Deferasirox induces VEGF in A549 and H1299 supernatant via up-regulation of HIF1a.

[Conclusion] Iron controlled treatment can be novel therapeutic agent and biomarker of Bevacizumab.

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 4247. doi:10.1158/1538-7445.AM2011-4247
Abstract 4325: Localization and expression of osteopontin in a chitosan-treated ovarian cancer cell line, SKOV-3
Authors: Regisford E. Gloria C. , Stone Julia M. , Carson Laura E. , Kelly-Brown Cordella , Traisawatwong Pasakorn , Hicks Antoine
Published: June 2012

Ovarian cancer is the deadliest gynecological malignancy and despite current treatments, such as chemotherapy and radiotherapy, reoccurrence of this deadly malignancy is high. Although elevated levels of osteopontin (OPN), a phosphorylated glycoprotein, are always present in the ovaries, recent studies have implicated osteopontin (OPN) protein as a novel biomarker for ovarian cancer. Early detection of ovarian cancer is imperative, and furthermore, novel treatments with fewer adverse effects than current treatments of chemotherapy and radiation, are currently being investigated. Chitosan, a natural polysaccharide that is biodegradable and biocompatible, may serve as a potential alternative treatment for ovarian cancer, and may function by regulating OPN expression. Therefore, the objective of this study was to determine the effects of chitosan on the expression and localization of OPN in SKOV-3 cells. It was hypothesized that the expression of OPN protein will be down-regulated in a dose-dependent manner. To test this hypothesis, SKOV-3 cells were treated with 0, 50, 100, 250, and 500, ng/mL of chitosan for 48 hours. An MTT assay was performed to determine cell viability and proliferation rate. Cells were harvested; total protein was isolated and then subjected to SDS-PAGE and Western blot analysis. The localization of OPN was determined using immunocytochemistry, after chitosan treatment of SKOV-3 cells on two-chamber slides for 48 hours. The expression of OPN decreased in SKOV-3 cells in a dose-dependent manner. In addition, cell proliferation tended to decrease with increasing dose of chitosan. These data indicate that chitosan may inhibit SKOV-3 cell proliferation by decreasing OPN production and secretion.

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 4325. doi:10.1158/1538-7445.AM2011-4325
Abstract 4330: Endometrial hyperplasia resulting from myeloid-specific targeting of the tumor suppressor APC
Authors: Kitchen-Goosen Susan M. , Alberts Art
Published: June 2012

The Adenomatous polyposis coli (APC1) tumor suppressor is a multi-function protein with a central role in Wnt-regulated cell growth and differentiation. Additionally, APC functions to modulate microtubule and microfilament dynamics to establish polarity in epithelia during directed cell migration and asymmetric cell division. APC frequently collaborates with the canonical Diaphanous-related formin, mDia1 (DIAPH1), which also binds microfilaments and microtubules to modulate cytoskeletal dynamics. APC1 and DIAPH1 are genetically linked on chromosome 5q in a region frequently disrupted in myeloproliferative neoplasms (MPNs). mDia1 knockout mice develop an age-dependent outgrowth of myeloid-lineage cells. The physical and functional linkage of APC and mDia1 as 5q tumor suppressors and in cytoskeletal remodeling led us to explore the role of APC in tumor suppression in myeloid-derived cell lineages affected by mDia1 knockout.

To test this hypothesis in a genetic model, we created a mouse in which APC was knocked out in myeloid lineage cells by breeding the Apc-flox mice with mice expressing Cre recombinase under the control of lysozyme M (LyzsM) gene promoter. The resulting phenotype was unexpected and 100% penetrant: Female mice developed cystic endometrial hyperplasia and were unable to sustain pregnancy. Embryo implantation occurred, but embryos were resorbed mid-gestation irrespective of progeny genotype. Two hypothesized mechanisms to explain the initiation of endometrial hyperplasia are being pursued. The first hypothesis is that loss of APC impairs the homeostasis between growth and differentiation of myeloid lineage stem/progenitor cells that transdifferentiate into uterine epithelia. An alternate hypothesis is that loss of APC expression affects the inflammatory response of resident uterine macrophages that lead to defects in epithelial morphology. In either case, these genetic studies imply that APC gene defects in myeloid lineage cells may be an initiating event in endometrial cancer.

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 4330. doi:10.1158/1538-7445.AM2011-4330
Abstract 4331: Characterization of targeted overexpression of Claudin-1 transgenic mice
Authors: Pope Jillian L. , Bhat Ajaz , Krishnan Moorthy , Beauchamp R. D. , Singh Amar , Dhawan Punita
Published: June 2012

Disruption of cell-cell junctions and changes in the expression of junctional proteins is a critical event in the progression of cancer. We have previously reported a tumor stage specific increase in the expression of a tight junction protein, Claudin-1, in colorectal cancer and its association with tumor-growth and metastasis. Furthermore, our studies have shown that in colon cancer cells, claudin-1 expression is regulated by a complex interaction between APC, Smad4 and β-catenin, three important regulators of colorectal carcinogenesis. We wanted to investigate the consequence of overexpression of claudin -1 in vivo specifically in the colon.

We have generated a transgenic mouse model in which Claudin-1 is expressed under the control of the villin promoter. The villin promoter is expressed in the adult intestinal epithelium and has been frequently used for intestinal-specific overexpression of several genes of interest. Western blot analysis demonstrated a robust expression of claudin-1 in the colon, small intestine and cecum of transgenic mice while low levels of expression were present in wild type mice. These results are confirmed with immunohistochemistry analysis where claudin-1 was largely expressed on the membrane in the same tissues. Colonic permeability was assayed using FITC-dextran, however, there was no significant difference between the flux of FITC-dextran in wild-type and transgenic mice. Real-Time PCR analysis for inflammatory cytokine expression showed a decrease in IL-10 mRNA in transgenic mice compared to that of wild-type mice. Further, treatment with Dextran Sodium Sulfate (DSS) to induce colitis showed a significant decrease in body weight of claudin-1 transgenic mice as compared to wild type mice on DSS. An increase in disease severity was observed in transgenic mice as determined by histology scoring, as well as a significant difference in the ratio of colon weight to colon length. Claudin-1 expression in transgenic mice is localized to the membrane, where it maintains its normal function. Further experiments with this model include combined treatment with DSS and AOM to examine the effect of Claudin -1 overexpression in inflammation-induced-tumorigenesis as well as performing crosses with sporadic models of tumorigenesis involving mutations in the APC gene, to examine the role of claudin-1 upregulation in tumor formation and progression.

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 4331. doi:10.1158/1538-7445.AM2011-4331
Abstract 4368: Targeting of head and neck cancer stem cells using salinomycin
Authors: Kuo Selena Z. , Blair Katherine J. , Abhold Eric L. , Kiang Alan , Haas Martin , Wang-Rodriguez Jessica , Ongkeko Weg M.
Published: June 2012

Cancer stem cells are believed to play a crucial role in cancer recurrence due to their resistance to conventional chemotherapy and capacity for self-renewal. Gupta and colleagues recently reported that salinomycin, a livestock antibiotic, selectively targets breast cancer stem cells. In our study we sought to determine the effect of salinomycin on head and neck squamous cell carcinoma (HNSCC) stem cells. We observed that salinomycin inhibits self-renewal capacity in HNSCC stem cells, as shown by a dose-dependent decrease in sphere formation. In addition, treatment with salinomycin resulted in a decrease in mRNA expression of the stem cell markers CD44 and BMI-1, both of which are critical for maintaining tumorigenicity in HNSCC. In combination with the chemotherapeutic agents cisplatin and doxorubicin, salinomycin synergistically killed HNSCC cancer stem cells more effectively compared to either drug alone, as demonstrated by cell proliferation and TUNEL assays. High-throughput analysis revealed a set of differentially expressed microRNAs in salinomycin-treated HNSCC stem cells, warranting further investigation into the possibility of microRNA-mediated pathways that are regulated by salinomycin. Taken together, our findings indicate promise for using salinomycin or related derivatives as a novel treatment for HNSCC, although further studies are needed to confirm efficacy in vivo.

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 4368. doi:10.1158/1538-7445.AM2011-4368
Abstract 4370: Network-based signatures for drug repositioning and combination for the breast tumor initiating cells
Authors: Jin Guangxu , Zhao Hong , Cong Yang , Fu Changhe , Chang Jenny , Lewis Michael , Wong Stephen
Published: June 2012

Understanding the signaling mechanisms of breast tumor initiating cells (TIC) is important to design efficient therapeutic and management strategies of breast cancer. We present a network-based signature and a comprehensive signaling map for identification of candidates of drug repositioning and combinations for breast TIC. The network-based signature is based on an extended concept of network motifs, known as cancer-signaling bridges (CSBs), which can be used to expand the cancer drug-targets of known signaling pathways. We use the profiles of TIC derived from CD44+/CD24-/low breast cancer cells and mammospheres (MS) cells to establish network-based signatures. Facilitated by the signaling pathways that are highly connected with CSBs, e.g., MAPK, NOTCH, ECM-receptor, Jak-STAT, and Wnt, we first identify the high-confidence signaling paths automatically chosen out of CSBs by two scoring systems, namely, Differential Expression Score (DES) and Signaling Pathway Score (SPS). The high-confidence signaling paths are used to build the network-based signature that characterizes the breast TIC. The network-based signature for CD44+/CD24-/low cells is composed by 140 proteins and 132 protein-protein interactions and that for mammospheres contains 153 proteins and 119 protein-protein interactions. The FDA-approved drugs whose targets are included in the network-based signatures are repositioned to breast TIC as the candidates for the repositioning. Furthermore, we curate a comprehensive signaling map for breast TIC by using the available signaling transductions in BioCarta, KEGG, and IPA and include seven signaling pathways, PI3K/AKT, JAK/STAT, Notch, HH, Wnt, P53, and ECM. The signaling map enables us to further refine the repositioning candidates and eventually propose combination candidates by using the cross-talks of signaling pathways in the map. Using this mapping approach, the drugs targeting on TNF, KDR, and IKBKB, such as sunitinib, Arsenic trioxide, and Atorvastatin, are repositioned for treating breast TIC, while the crosstalks between pathways such as Notch+hedgehog and Hedgehog+wnt+PI3K are used to identify drug combination candidates from the target combinations, TNF+KDR, and TNF+IKBKB.

Acknowledgements: This research is partially funded by NIH ICBP U54CA149169. The authors would also appreciate the discussion and advice of Jeff Rosen and David Tweardy from Baylor College of Medicine during the generation of comprehensive signaling map for breast TICs.

Guangxu Jin and Hong Zhao contribute to this work equally.

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 4370. doi:10.1158/1538-7445.AM2011-4370
Abstract 4375: Cancer protection by pregnancy – a link with microchimeric fetal/maternal stem cells in a stromal niche: Breast cancers are hypochimeric, but the HER2-subtype is hyperchimeric
Authors: Dhimolea Eugen , Denes Viktoria , Aziz-Zaman Sonya , Geck Peter
Published: June 2012

The PURPOSE of our studies was to investigate the origin and cancer relations of embryonic/fetal stem cells in breast cancer.

HYPOTHESIS. During pregnancy the documented exchange of fetal and maternal stem/progenitors establish long-term integration in the recipient organism. We propose that the breast is a potential niche and this new stem cell pool may affect breast cancer.

OBJECTIVES. (i) To work out more sensitive methods to detect Y- and X-chromosomal markers from embedded, formaldehyde-fixed tissue samples; (ii) To find the tissue compartment for the long-term microchimerism niche in the breast; and (iii) to collect quantitative chimerism data to investigate non-chimeric, hypochimeric, microchimeric and hyperchimeric contribution to cancer.

EXPERIMENTS. We developed new systems for Y-signal detection (single-amplicon/multiple-target analysis) and X-haplotype analysis with significantly increased sensitivity from low-integrity samples. We analyzed 55 normal samples representing stromal, ductal and lobular tissues and 102 invasive ductal breast cancer samples. Quantitative amplifications were performed following recent MIQE recommendations. We used fluorescent in-situ hybridizations (FISH) to confirm microchimerism. The data were analyzed by extensive statistical analyses and only data with statistical significance are presented.

RESULTS. We found 19.6% microchimerism in cancer samples, in contrast to 40% in normal tissues. The results confirm that hypochimerism promotes but microchimerism protects from cancer. We show for the first time that the long-term niche for post-pregnancy microchimeric (male) cells is the mammary stroma but they don't integrate into the ducts, the site of most cancers. Based on the findings we propose that breast cancer protection by pregnancy may be a stromal function and involves fetal microchimerism. Unexpectedly, we also found that a cohort of breast cancers show runaway microchimerism with significantly increased male-cell content. This is the first report that hyperchimerism correlates with the HER2-subtype and unfavorable prognosis.

CONCLUSIONS. We developed a new assay and our studies demonstrate previously unrecognized significant stromal integration of fetal/maternal stem cells in protection from breast cancer. We also found, however, that excessive microchimerism promotes cancer and correlates with the HER2-subtype.

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 4375. doi:10.1158/1538-7445.AM2011-4375
Abstract 4394: Identification of molecular determinants of response to GSK1120212B, a potent and selective MEK inhibitor, as a single agent and in combination in RAS/RAF mutant non-small cell lung carcinoma cells
Authors: Liu Li , Hong Shi , Zhang Vivian , Gilmer Tona
Published: June 2012

MEK, a downstream effector for RAS, RAF and receptor tyrosine kinases, represents a promising target for treatment of RAS/RAF mutated Non Small Cell Lung Carcinoma (NSCLC). GSK1120212B is a highly potent and selective oral MEK inhibitor currently in phase II clinical trials. To identify markers of response to GSK1120212B and to generate hypotheses for effective combination therapies, we characterized baseline signaling pathways and gene expression profiles in a panel of 29 NSCLC lines with RAS/RAF mutations and determined their sensitivity to GSK1120212B, erlotinib (EGFR inhibitor), everolimus (mTOR inhibitor), docetaxel and the combination of GSK1120212B with these agents. Inhibition of cell growth and induction of apoptosis were determined after 72 and 24 hours of treatment, respectively. GSK1120212B potently inhibited cell growth (IC50≤ 50 nM) in 13/29 (45%) of the lines and induced apoptosis in 8/29 (28%) of cell lines. We also identified genes such as TGFA, EREG, LGALS3, CTNND1, ARHGEF3, KLF4, RPH3AL, STN2, TWIST2 and FOXC1 whose expression levels were highly associated with the response to GSK1120212B. Cell signaling analyzed at baseline demonstrated that sensitivity to GSK1120212B was associated with high levels of pMEK, low levels of pAKT/pFOXO3a and wild-type PTEN, and resistance was associated with high levels of pAKT/pFOXO3a or mutations of PTEN/PIK3CA/CTNNB1 in RAS/RAF mutant lines. These results suggest combination approaches for cell lines resistant to single agent GSK1120212B. Erlotinib or everolimus alone showed little activity in RAS/RAF mutant lines. The combination of GSK1120212B and erlotinib was synergistic and/or enhanced cell growth inhibition in 6 out of 10 RAS/RAF mutant lines tested, three of which had HER1 amplification or over-express TGFA and EREG. The combination of GSK1120212B and everolimus enhanced cell growth inhibition in 25/29 (86%) of cell lines and increased apoptosis in 12/28 (43%) of RAS/RAF mutant lines. Intriguingly, the RAS/RAF mutated lines that were sensitive to GSK1120212B alone as measured by induction of apoptosis were less responsive to docetaxel. Further, the combination of GSK1120212B and docetaxel was synergistic or increased apoptosis in 20/28 RAS/RAF mutated lines. Our results provide a strong rationale for testing GSK1120212B and these combinations in the clinic and warrant further investigation of the gene expression signatures and genetic mutations in patients to ultimately improve treatment outcome in RAS/RAF mutated NSCLC.

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 4394. doi:10.1158/1538-7445.AM2011-4394
Abstract 4406: Multiple organic anion transporting polypeptides (OATPs) contribute to the in vitro and in vivo disposition of docetaxel
Authors: Ho Richard H. , Leake Brenda F. , Teft Wendy , Kim Richard B.
Published: June 2012

Docetaxel is one of the most widely used antineoplastic agents and is approved for the treatment of breast, lung, ovarian, prostate, gastric, and head and neck cancers. It is a semisynthetic taxane derived from an extract of the needles of European yew tree (Taxus baccata). Docetaxel promotes the assembly of microtubules and stabilizes their formation by inhibiting depolymerization and cell death results from inhibition of normal microtubule processes. While it is known that docetaxel is a substrate of efflux transporters such as P-glycoprotein (P-gp), less is known regarding transporters that could mediate its cellular uptake. Accordingly, an array of drug uptake transporters, including organic anion, organic cation and bile acid transporters, were evaluated for radiolabeled docetaxel transport utilizing a recombinant vaccinia-based method in a heterologous cell system. Screening transport experiments and kinetic analysis in HeLa cells demonstrated that multiple organic anion transporting polypeptides (OATPs), including OATP1A2, OATP1B1, and OATP1B3, were capable of docetaxel transport in vitro. In addition, after intravenous administration, in vivo docetaxel transport experiments in Oatp1b2-/- mice (ortholog to hepatic OATP1B1/OATP1B3) revealed ∼5-fold higher plasma concentrations of docetaxel in Oatp1b2-/- mice versus their wild-type counterparts. The liver to plasma ratio of docetaxel was ∼3-fold increased in wild-type mice versus Oatp1b2-/- mice. Furthermore, the pharmacokinetic profile of docetaxel is characterized by substantial interindividual variability, with an up to tenfold difference in drug clearance, even in patients with normal hepatic function. A panel of single nucleotide polymorphisms (SNPs) in OATPs was assessed for docetaxel transport. Multiple SNPs in SLCO1A2, 1B1 and 1B3 were associated with impaired docetaxel transport in vitro. In conclusion, multiple OATPs were capable of transporting docetaxel in vitro and in vivo, suggesting these transporters are important to docetaxel disposition. A number of OATP variants were associated with significantly impaired docetaxel transport, which may have important toxicological and therapeutic ramifications.

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 4406. doi:10.1158/1538-7445.AM2011-4406