bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 322: Loss of Y chromosome in urological tumors
Authors: Minner Sarah , Kilgué Adisa , Stahl Phillip , Handel Christoph , Höppner Wolfgang , Wagner Walter , Dahlem Roland , Rink Michael , Eichelberg Christian , Schlomm Thorsten , Terracciano Luigi , Moch Holger , Fisch Margit , Simon Ronald , Sauter Guido
Published: February 2011

Introduction and Objective

Y chromosome losses have been associated with various malignant tumors including bladder cancer, kidney cancer and prostate cancer. Conflicting data exist on the potential prognostic role of Y chromosome losses in these urological malignancies and on whether Y losses could be age-related. The aim of this study was to further clarify the potential relevance of Y chromosome losses in these tumors with respect to tumor phenotype, clinical outcome and patient age.

Methods

Preexisting tissue microarrays (TMA) of bladder (573 tissue samples from male patients), kidney (447 tissue samples from male patients) and prostate cancer (3261 tissue samples) with long-term clinical follow-up data were used in this study. Y chromosome losses were analyzed by dual-labeling FISH (fluorescence in situ hybridization) using centromere Y and X probe.

Results

Y losses were seen in 22% of bladder cancers. There was no significant age difference between patients having tumors with (67.4 +/-2.1years) or without (67.3 +/-1.2 years) Y losses (p=0.6294). Y losses were unrelated to tumor grade (p=0.7831) and stage (p=0.6140). There was no correlation between Y losses and survival in 224 invasive urothelial cancers (pT2-4; p=0.2324). Y losses were not associated with an increased risk for recurrences in 197 analyzed pTa tumors (p=0.7649) or increased progression risk in 76 analyzed pT1 tumors (p=0.4582). In kidney cancer Y losses were seen in 23% of cases. Likewise, there was no significant age difference between patients having tumors with (59.2 +/-3.6 years) or without (61.2+/-1.5 years) Y losses (p=0.5873). Y losses were unrelated to tumor grade (p=0.3111) and tumor stage (p=0.2511). There was also no correlation between Y losses and survival (p=0.7431). Remarkably, Y losses were seen in only 13 of 2826 (0.5%) analyzable cases of prostate cancers. The number of Y chromosome losses was too low for further statistical analyses.

Conclusions

These data show, that Y losses are frequent in urinary bladder cancer and kidney cancer, but lack association with tumor phenotype, clinical outcome and patient age. While the mechanism for Y losses in these cancers remains unclear, our data do not suggest clinical significance. In contrast to bladder and kidney cancer, Y chromosome losses were almost absent in prostate cancer. This may suggest that prostate epithelial cells are highly dependent on proper function of at least one chromosome Y gene.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 322.
Abstract 3222: Telomerase expression is upregulated in response to severe oxidative stress
Authors: Sharif Wallace , Butts DeMario , Brown Jamelle
Published: February 2011

Telomerase is aberrantly expressed in 90% of all cancers, and is well characterized for its ability to extend the ends of chromosomes, the telomere. Although telomerase expression is predominantly associated with its ability to extend telomeres, there have been recent findings that suggest that telomerase has multiple cellular functions independent of its catalytic ability, and some of those functions help promote cell survival in response to severe oxidative stress. To date, telomerase expression as been implicated in aiding a cell's response to double strand breaks in DNA, mitochondria dysfunction, inhibition of apoptosis, and the promotion of cell proliferation. The protein component of telomerase, TERT, is also thought to specifically enhance cell viability in response to the hypoxic microenvironment of a tumor. These protective roles of TERT could be vital in promoting cell survival during severe hypoxia. Telomerase expression is upregulated by the kinase Akt, which is in turn induced by the neuregulin/ErbB signaling pathway. Neuregulin is a glycoprotein that is necessary for smooth muscle and neuronal development. Neuregulin activates the ErbB tyrosine kinase family, which promotes cell survival, proliferation, and differentiation. Recently, neuregulin has been shown to be upregulated in response to severe hypoxia, and may reduce the inflammation response to oxidative stress. The mechanism of neuregulin protection is unclear, though microarray data shows that neuregulin expression decreases the expression of several pro-inflammatory cytokines. We hypothesize that neuregulin can regulate the expression of telomerase during severe oxidative stress, and that telomerase expression promotes cell survival during hypoxia. Inducing oxidative stress and hypoxia in differentiated rat neuroblastoma cells caused significant increases in telomerase expression. The addition of physiologic levels of neuregulin was able to induce telomerase expression, indicating that telomerase activation could play an important role in the cellular response to oxidative stress.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3222.
Abstract 3225: Cellular and in vivo models for the analyses of B-Raf and c-Src inhibitors
Authors: Ehlert Jan E. , Mutschler Bettina , Müller Melanie , Lingnau Andreas , Hoffmann Steffen , Kubbutat Michael HG
Published: February 2011

Cytosolic kinases c-Src and B-Raf represent proto-oncogenes of which mutant or overexpressed variants have been shown to cause cancer. Efforts are taken to develop inhibitors specifically addressing these kinases, some of which have already been approved such as Src inhibitor Dasatinib and BRaf inhibitor Sorafenib. In the current study we have generated cellular systems to allow for the analyses of Src and BRaf inhibitors in kinase specific cellular phosphorylation assays as well as in subcutaneous in vivo models. For both kinases we are introducing in vivo models allowing for regulatable kinase activity and thereby regulatable, kinase dependent tumor growth.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3225.
Abstract 3230: Immortalization of human small airway epithelial cells as a model for studying lung tumor transformation
Authors: Gao Boning , Huang Chunxian , Sullivan James , Raso Gabriela , Wistuba Ignacio , Shay Jerry , Minna John
Published: February 2011

Normal human lung epithelial cells that can be genetically manipulated to study the various steps in lung cancer pathogenesis and stem cell biology would be of great utility and also provide isogenic strains to develop and test new diagnostic (biomarkers), preventative and therapeutic strategies for lung cancer. Previously we immortalized normal proximal human lung bronchial epithelial cells (HBECs) for studying lung tumors arising in the central compartment of the lung and now have over 40 of these strains (growing in defined KSFM media) including many isogenic derivatives with various oncogenic changes (Can Res 2004, Can Res 2006). We have extended this by immortalizing normal distal (small airway) epithelial cells that serve as models for studying tumors arising in the peripheral compartment of the lung. Using Cdk4 and hTERT exogenous expression vectors (as used for HBECs) and SAGM defined media routinely immortalized (N= 15) human small airway epithelial cells (HSAECs) from peripheral lung. These HSAEC strains are highly enriched (>50%) for cells with stem like properties (e.g. Aldehyde dehydrogenase positivity), express basal cell markers but not Clara cell, type 1 or type 2 differentiation markers., However, HSAECs can be induced to express Clara cell, type 1 or type 2 cell markers when they are grown in matrigel (where they form cyst like structures which express such differentiation markers) and under other conditions favoring differentiation (such as insulin depletion, Ca2+ addition). HSAECs show anchorage dependent growth, do not form colonies in soft agar, and are non-tumorigenic in NOD/SCID mice. However, HSAECs can be genetically manipulated by introducing oncogenes (such as KRASV12, missense p53 and other oncogene combinations) which now allow HSAECs to progress towards a transformed phenotype (e.g. formation of soft agar colonies), and selection of a subset of HSAEC(KRAS, p53) large soft agar colonies identifies sublines that can now form poorly differentiated carcinomas. Thus, we have developed conditions to routinely immortalize lung epithelial cells derived from distal small airways (HSAECs) which grow in defined media, exhibit stem cell like characteristics, and can be genetically manipulated to give rise to poorly differentiated cancers.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3230.
Abstract 3235: Creation of novel rat models for cancer research using zinc finger nuclease (ZFN) technology
Authors: McCoy Aaron M. , Ji Diana , Carbery Iara , Cui Xiaoxia , Weinstein Edward
Published: February 2011

The rat resembles human physiology more closely than the mouse, partially because of its larger size. Rats are the preferred model for many disease fields, including cancer research, such as studies in breast and prostate cancers and bone metastasis. However, the lack of convenient experimental tools to manipulate the rat genome has largely limited the use of rat models, until the recent creation of the first targeted knockout rats by using zinc finger nuclease technology (Geurts et al., 2009).

At SAGE Labs, we are uniquely positioned to create rat knockout models for all major disease categories. In regards to cancer research, we have developed p53 knockout rats, which may be used to significantly shorten the time required for carcinogenicity studies in the same capacity as the p53 knockout mice have been used since the mid-nineties. We are also in the process of creating a suite of immunocompromised rats, such as knockouts of RAG1, RAG2, and DNAPK, which may be used, for example, in the generation of xenografts for metastasis studies and bone marrow transplantation for cancer therapy. In addition, we have developed a set of toxicology models, including knockouts of Mdr1a, PXR, BCRP, Mrp1 and Mrp2, which can benefit studies on cancer drug delivery and metabolism. Model generation, their applications and future models will be discussed. Together, our goal is to help advance cancer research with the creation of novel rat models.

Geurts, A. et al. Knockout rats via embryo microinjection of zinc-finger nucleases. Science 325, 433 (2009)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3235.
Abstract 3237: Establishment of a new invasive urinary bladder cancer model using human c-Ha-ras proto-oncogene transgenic rats
Authors: Wei Min , Yamano Shotaro , Ishii Naomi , Kakehashi Anna , Obo Yumi , Wanibuchi Hideki
Published: February 2011

To establish animal models for identifying mechanisms of urinary bladder cancer (UBC) invasion, male human c-Ha-ras proto-oncogene transgenic rats (Hras 128) were divided into 5 groups and treated with 0.05% N-butyl-N-(hydroxybutyl)nitrosameine (BBN) in drinking water and/or 0.1% phenylethyl isothiocyanate (PEITC) in diet, respectively, as follows: BBN (8 wks)→PEITC (8 wks); PEITC (8 wks)→BBN (8 wks); PEITC alone (16 wks); BBN alone (16 wks); non-treatment. At the end of week 16, incidences of invasive UBC in BBN→PEITC and BBN alone groups were 77% (10/13 rats) and 56% (5/9 rats), respectively. The average number of invasive UBC in BBN→PEITC and BBN alone groups were 1.3 ± 0.9 and 0.7 ± 0.7/rat, respectively. Both incidence and numbers of invasive UBC tended to be higher in BBN→PEITC group compared to the BBN alone group. Furthermore, non-invasive UBC were also found in all rats of above 2 groups. No tumors were found in other groups. p53 positive staining were observed in 58% of invasive UBC and 9% of non-invasive UBC in BBN→PEITC group that is similar to positive incidences in human UBCs. These findings indicated that treatment of Hras 128 rats with BBN followed by PEITC induced high incidences of both non-invasive and invasive UBC, and therefore provide a useful comparison model for explore the mechanisms of UBC invasion.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3237.
Abstract 324: A NUT-variant midline carcinoma showing a complex karyotype
Authors: Thompson Katherine , Beesley Alex H. , Baker Elizabeth , Grumball Tanya , Kennedy Dagmara A. , Murch Ashleigh R. , Philips Marianne B. , Charles Adrian K. , French Chris A. , Kees Ursula R.
Published: February 2011

NUT-midline carcinomas (NMC) are rare, almost uniformly lethal malignant tumours arising in the midline organs. Originally diagnosed predominantly in the young, they have since been identified in people ranging in age from 3 to 78 years. The tumours are typically very aggressive in behaviour, with an average survival of only nine months. NMCs are typified by a single, conserved underlying genetic translocation t(15;19)(q14;p13.1), that results in a fusion of the BRD4 and NUT genes. There are a few cases described in which NUT is fused to a different gene (primarily BRD3) and these are known as NUT-variants. It is very rare however, for translocations other than the primary one, to be present. We present an NMC case with a far more complex karyotype, with the translocations t(6;19)(q14;p13.1) and t(1;18;7)(q42;q11.2;q21) present in all cells, as well as approximately 10% of cells demonstrating t(12;15)(q13;q22). Initial FISH experiments, using break-apart probes, established a translocation of the 19p13.1 (BRD4) region, and demonstrated no translocation of the 15q14 (NUT) region. Subsequent experiments, using BRD4 specific probes in a cell line derived from the patient specimen, have shown that BRD4 was inserted into chromosome 15, in a region proximal to the NUT gene. We are currently determining whether BRD4 was fused directly to NUT through a cryptic translocation in this case. Illumina Human-Cyto-SNP-12 BeadChip array analysis has also shown that a 3.3MB region of 6q14 (adjacent to the region translocated onto chromosome 19) was deleted in the tumour cells. The array indicated no deletions or duplications at the sites of any of the other translocations. Given the aggressive nature of these tumours, a better understanding of the underlying genetic causes and precipitating factors of the disease processes remains imperative to improving the outcome for such patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 324.
Abstract 3241: Multipotential PTEN−/− p53−/− prostate tumor initiating cells give rise to adenocarcinoma which displays epithelial-to-mesenchymal transition, metastasis, and androgen independent growth
Authors: Pierce Rachel M. , Martin Philip L. , Kelly Kathy
Published: February 2011

Determining the cell lineage and differentiation potential of tumor initiating cells is fundamental to understanding mechanisms of progression and metastasis. We describe the differentiative, tumorigenic, and metastatic properties of primary, transplanted secondary, and clonally-derived tumors in a model of prostate cancer initiated by prostate epithelial cell specific deletion of the tumor suppressors, PTEN and p53. Although adenocarcinoma is the majority tumor type that develops in this mouse model, other histological tumor types, including a late developing sarcomatoid carcinoma were observed. The heterogeneity displayed in transgenic mouse prostate tumors may be due to the transformation of multiple lineage specific tumor initiating cells, or a common tumor progenitor/stem cell capable of plasticity in differentiation potential. Orthotopic transplantation of PTEN−/− p53−/−prostate carcinoma cells was undertaken to determine whether a single tumor initiating cells is capable of giving rise to the diversity of tumor phenotypes observed in the parental prostate carcinoma. Direct orthotopic transplantation of mixed populations of PTEN−/− p53−/− tumor cells favored the development of adenocarcinomas, while transplantation of cells grown as protospheres in matrigel favored the development of basal/squamous carcinomas. This indicates a developmental plasticity in transplantable tumor initiating cells that can be influenced by extracellular matrix components. Transplantation of clonal cell lines that were derived from orthotopic carcinomas produced adenocarcinomas with lesser amounts basal/squamous carcinoma with one line undergoing EMT to form sarcomatoid carcinomas. Lung and lymph node metastases were common. Tumor growth, metastasis, and response to androgen deprivation were assayed with in-vivo bioluminescent imaging. The orthotopic tumors recapitulated the diversity of tumor phenotypes observed in the probasin-cre, PTEN fl/fl p53 fl/fl transgenic donor mouse including: adenocarcinoma, spindle cell carcinoma, urothelial carcinoma, and basal/squamous carcinoma. In a pattern that mimics a phenotype observed in metastatic human prostate carcinoma, CK8+ adenocarcinomas displayed heterogeneous labeling for the neuroendocrine marker synaptophysin. Our data suggests that a multi-potential CK8+/CK5+ tumor stem cell can be a progenitor for CK8+ adenocarcinoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3241.
Abstract 325: ITCH is a putative target for a novel 20q11.22 amplification detected in anaplastic thyroid carcinoma cells by array-based comparative genomic hybridization
Authors: Ishihara Takaya , Tsuda Hitoshi , Hotta Akiko , Kozaki Ken-ichi , Yoshida Akira , Yoshimura Hiroshi , Ito Koichi , Imoto Issei , Inazawa Johji
Published: February 2011

Anaplastic thyroid carcinoma (ATC) is one of the most virulent of all human malignancies, with a mean survival time among patients of less than a year after diagnosis. To date, however, cytogenetic information on this disease has been very limited. Significant genomic copy-number alterations, such as amplifications and homozygous losses, are useful landmarks for identifying oncogenes and tumor-suppressor genes (TSGs), respectively. We have performed bacterial artificial chromosome (BAC) array-based comparative genomic hybridization (array-CGH), which allows the high resolution, high throughput and quantitative copy-number analysis throughout the genome, in various cancers, and identified many novel cancer-related genes. In ATC, we have reported DUSP26 as a novel growth-promoting gene from an amplified region detected in ATC cell lines by the array-CGH using an in-house low-density cancer-targeting array (Yu Wei et al.,Oncogene 2007). In the course of a program to screen a panel of ATC cell lines for genomic copy-number aberrations using array-based comparative genomic hybridization, we identified a high-level amplification of the ITCH gene, which is mapped to 20q11.22 and belong to the HECT ubiquitin ligase family. The expression of ITCH was increased in four of 14 ATC cell lines (28.6%), including 8305C with a copy-number amplification of this gene, and six of seven primary cases (85.7%). Among the primary thyroid tumors, a considerable number of ITCH high expressers were found in ATC (40/45, 88.9%), papillary thyroid carcinoma (25/25, 100%), and papillary microcarcinoma (25/25, 100%). Furthermore, knockdown of ITCH by specific small interfering RNA significantly inhibited the growth of ITCH-overexpressing cells, whereas ectopic overexpression of ITCH promoted growth of ATC cell lines with a relatively weak expression. These observations indicate ITCH to be the most likely target for 20q11.22 amplification and to play a crucial role in the progression of thyroid carcinoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 325.
Abstract 3253: Genetic loci that contribute to aristolochic acid nephropathy and associated upper urothelial cancer
Authors: Rosenquist Thomas A. , Strockbine Penelope , Grollman Arthur P.
Published: February 2011

Aristolochic acids (AA) are nitrophenanthrene carboxylic acids found in all Aristolochia species which have been used historically as herbal remedies throughout the world. AA are the suspected causative agent for aristolochic acid nephropathy (AAN), Balkan Endemic Nephropathy (BEN), and so-called Chinese Herb Nephropathy (CHN). Hallmarks of these disease entities are proximal tubule damage and interstitial fibrosis leading to end stage renal disease. At least half of all patients also develop cancers of the upper urothelial tract (UUC). Aristolochic acid-DNA adducts generate signature A:T->T:A transversions in TP53. Such mutations are rare in sporadic urothelial cancers but predominate in UUC from patients with BEN/AAN/CHN.

We have developed a mouse model for AAN that has enabled us to screen for quantitative trait loci (QTL) that increase the susceptibility to AA in mice. Two hundred F2 mice obtained by breeding C57Bl/6J (an AAN-resistant strain) and DBA/2J (an AAN-sensitive strain) were treated with a single dose of AA. Four days later urine was analyzed for markers of kidney function. Mice were then euthanized and tissues were collected for histology and DNA was isolated for analysis of AA-DNA adducts and genotyping.

Four QTL were identified; a major QTL, Aanq1(Chr4, 46-71MB), a pair of interacting QTL, Aanq2(Chr3, 116-128MB), and Aanq3 (Chr9, 113-122MB), and a minor QTL that acts additively with Aanq1, Aanq4(Chr6, 124-135MB). Further analyses of the QTL regions and their human orthologs will be presented.

The project described was supported by Award Number P01ES004068 from the National Institute of Environmental Health Sciences.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3253.
Abstract 3286: Overexpression of cornulin in histologically normal adjacent tissue predicts occult nodal metastases in head and neck cancer patients
Authors: Weinberger Paul M. , Merkley Mark , Jackson Lana , Dynan William S.
Published: February 2011

As with many epithelial cancers, presence of nodal metastases in patients with Head and Neck Squamous Cell Carcinoma (HNSCC) predicts early recurrence and poor survival. Determination of occult metastases in the clinically N-zero (cN0) patient remains problematic and therefore prophylactic neck dissections are routinely performed for high risk cN0 patients. A cohort of 7 cN0 HNSCC patients who subsequently underwent planned neck dissection as part of their treatment was selected. Snap frozen tissue was enriched for tumor and adjacent normal tissue by laser capture microdissection. Total protein was extracted and separated by saturation-labeling 2D difference in-gel electrophoresis (2D-DIGE). Significance analysis of Microarray (SAM) method was used to evaluate for differential protein expression between patients with (n=3) and without (n=4) occult nodal metastases on neck dissection pathologic evaluation. Spot maps were generated for 14 gels (7 HNSCC, 7 normal) and relative abundance values calculated for each of 662 protein spots common to >90% of gels. A 5% false discovery rate and 2-fold change were used as criteria for significance in the defintion of candidate biomarkers. Protein spots meeting these criteria were then extracted, digested and analyzed by liquid chromatography and tandem mass spectrometry to obtain protein identifications. For tumor tissue, there was no significant difference between groups. There were considerable differences in protein expression for normal adjacent tissue between patients with and without occult metastases, with 60 spots meeting criteria for significance (FDR<0.05). There were 31 proteins underexpressed in occult metastases patients, with the top candidate being 11.9 fold lower in the occult metastasis group (adjusted q<0.001). There were 29 proteins overexpressed, with the top candidate being 6.6 fold higher in the occult metastasis group (adjusted q<0.001). This protein was identified as Cornulin, a 53 kDa calcium-binding protein of the s100 family recently identified as a novel HNSCC biomarker. Cornulin plays a known role as a cornification chaperone that is protective against acid stress in squamous epithelium. It is possible that elevated cornulin in surrounding normal tissue may reflect ongoing epithelial injury, predisposing developing tumors to aggressive behavior. In conclusion, we have identified overexpression of cornulin in adjacent normal tissue as a novel biomarker for tumors with occult metastases in the cN0 HNSCC patient population. Additionally, these data highlight the role that tumor-stroma interactions may play in the development of early nodal metastases.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3286.
Abstract 3294: The impact of occult nodal metastasis on the recurrence of esophageal squamous cell carcinoma
Authors: Morimoto Junya , Tanaka Hiroaki , Kubo Naoshi , Ohira Masaichi , Muguruma Kazuya , Noda Eiji , Amano Ryosuke , Inoue Toru , Yashiro Masakazu , Yamada Nobuya , Maeda Kiyoshi , Sawada Tetuji , Ishikawa Teturou , Hirakawa Kosei
Published: February 2011

Esophageal cancer (ESC) is an aggressive disease with a poor overall prognosis. Survival rate of patients with cStage II/III ESC who underwerrt surgical resection have been improved, whereas half of these patients frequently caused recurrences. It has been thought that the lymph node metastasis was deeply related to the relapse of ESC. However, clinical signification of micrometastases in lymph nodes remain unclear because of complexity in the procedure to detect micrometastasis. Here we report that the impact of micrometastasis in lymph nodes on the recurrence of ESC. The aim of this study was to confirm the relation between noccult nodal metastasis and recurrence of in resected cStage II/III ESC. A total of 98 patients with ESC resected at Osaka City University hospital between January 2000 and January 2007 were targeted. Micrometastasis was investigated with one-level cytokeratin immunohistochemistry(IHC)and routine histological assessment by hematoxylin-eosin sections. Judgment of micrometastasis was done with discovery of cluster cells. Uni-and multivariate analyses of the independent recurrect facters were performed using the Cox proportional hazards regression model.

A total of 6,172 lymph nodes(average 63) were examined from 98 patient, also 140 lymph nodes of micrometastasis(average 1.4) were detected. The micrometastasis was not possible to detect with routine histological assessment by hematoxylin-eosin sections. A clear difference of detected micrometastasis between relapse patients(45 patients,83%) and no relapse patient(15patients,34%) was admitted. In univariate analyses, curability(p=0.000),v facter(p=0.046),pN facter(p=0.026),pM facter(p=0.016),pStage(p=0.002),total metastatic lymph node count(p=0.000),total metastatic lymph node count added micrometastasis(p=0.000) were recurrence facters. In multivariate analyses, total metastatic lymph node count added micrometastasis were independent recurrence facters(Hazard ration 13.059,p=0.000) finally.

Therefore, the number of total metastatic lymph nodes with micrometastasis have pivotal impact on the recurrence of ESC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3294.
Abstract 3313: Snail and slug-mediated epithelial mesenchymal transition in lung cancer cells
Authors: Daisuke Tamura , Arao Tokuzo Arao , Nagai Tomoyuki Nagai , Aomatsu Keiichi , Sakai Kazuko , Kudo Kanae , Kaneda Hiroyasu , Matsumoto Kazuko , Furuta Kazuyuki , Fujita Yoshihiko , Kotani Yoshikazu , Nishimura Yoshihiro , Saijo Nagahiro , Nishio Kazuto
Published: February 2011

Epithelial-Mesenchymal Transition (EMT) has been classified as a unique process by which epithelial cells undergo remarkable morphologic changes characterized by a transition from epithelial cobblestone phenotype to elongated fibroblastic phenotype (mesenchymal phenotype) leading to increased motility and invasion. Accumulating evidence indicates that EMT-inducible transcription factors such as Snail homologues (Snail1, 2, and 3) and several basic helix-loop-helix factors Twist, are poor prognostic factors and associated with chemoresistance in oncology.

The aim of this study was to investigate the involvement of SNAI-mediated EMT and its malignant potential in lung cancer cells. We introduced SNAI1 or SNAI2 gene into A549 or PC-9 lung cancer cell lines and established stable cell lines A549/GFP, /SNAI1, /SNAI2, PC-9/GFP, /SNAI1 and /SNAI2. Overexpression of SNAI1 and SNAI2 clearly mediated the EMT-related morphological changes including the cell scattering and the elongation of cell-shape. Real-time RT PCR demonstrated that overexpression of SNAI1 and SNAI2 markedly down-regulated the mRNA levels of E-cadherin about 1/2 to 1/100 compared with GFP-expressing control cell lines. The mRNA upregulation of N-cadherin, fibronectin 1 and Vimentin, which changes are characteristic of EMT, were also observed. We are now investigating its oncogenic properties, gene expression changes by microarray analysis and chemoresistance using these stable SNAI1 or SNAI2 transfectants.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3313.
Abstract 3316: CD44High head and neck cancer cells demonstrate increased cell growth and chemotherapeutic resistance
Authors: Wu Arthur W. , Basak Saroj K. , Lai Chi , Veena Mysore S. , Wang Marilene B. , Srivatsan Eri S.
Published: February 2011

Background: Head and Neck Squamous Cell Carcinoma (HNSCC) is a tumor made up of a heterogeneous population of cancer cells. Differences in individual cell phenotype may extrapolate to clinical behavior at the tumor level. CD44 is a cell surface marker highly expressed in putative cancer stem cells. Here, we demonstrate that fluorescence activated cell sorting (FACS) for the CD44 cell surface marker isolates a population of HNSCC cells with increased growth rate, colony forming ability, and chemoresistance. Methods: FACS was used to isolate CD44High and CD44Low cells from the HNSCC cell line SCC1. CD44High, CD44Low and unsorted cells were compared with respect to cell growth using the MTT assay for 8 days. They were also seeded into agar and grown for 6 weeks to determine colony forming potential. Sorted and unsorted cells were also compared for their sensitivity to cisplatin (20 µM). Live cells were measured using the MTT assay and compared to untreated cells at the same time point in order to calculate relative growth rates. Additionally, SCC1 cells were analyzed using FACS with and without cisplatin to determine changes in the population of CD44High cells. Primary tumor specimens and normal controls were analyzed for various cancer stem cell markers via immunohistochemistry. Results: CD44High SCC1 cells grew the fastest in culture with CD44Low cells growing significantly slower. Unsorted cells had a similar growth rate to CD44High cells, which is not surprising given that CD44High cells made up roughly 60% of the total population. CD44High cells in soft agar developed significantly more colonies with a larger average colony size after 6 weeks in culture compared to unsorted and CD44Low cells. CD44High SCC1 cells also were less sensitive to cisplatin with no difference in cell growth between treated and untreated cells. Using FACS to analyze unsorted cells after treatment with cisplatin, we observed enrichment in the CD44High cell population. Control non-cancer specimens demonstrated expression of p16 with low levels of BMI and CD44 on immunohistochemical analysis. Poorly differentiated HNSCC tumors demonstrated absence of p16, high BMI, and high CD44. Conclusion: The cell surface marker CD44 is expressed in a subset of HNSCC cells that have higher growth rates, tumorogenicity, and resistance to chemotherapy. These HNSCC stem cells may be the reason for resistance and recurrence. Understanding the mechanism behind their specific phenotype will be valuable in advancing treatments for HNSCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3316.
Abstract 3317: Study of response in neural stem cells after cerebral irradiation
Authors: Kong Doo-Sik , Joo Kyung Min , Nam Do-Hyun
Published: February 2011

Objective: Ionizing irradiation causes cell death but also birth and migration of new neurons within sites of cerebral damage. The cellular environment that induces neuronal regeneration and migration after irradiation has not been defined yet. The aim of this study was to evaluate the neuronal response after irradiation injury induced by gamma knife radiosurgery.

Methods: We used a model of long-distance migration of newly transplanted neural stem cells from the subependymal zone on the other hemisphere to that on the ipsilateral side after irradiation using gamma knife radiosurgery for the purpose of defining the cellular cues that induce neuronal regeneration after CNS injury.

Results: In vitro and in vivo studies showed that irradiation induced neurogenesis from transplanted GFP-tagged neural stem cells and migration of newly born neurons into a unique neurovascular niche. Within this neurovascular niche, newly born, immature neurons were closely associated with the remodeling vasculature. The relationship between neurogenesis and angiogenesis is causally linked through vascular production of CD31 and VEGF receptor-2.

Conclusions: These experiments defined a novel brain environment for neuronal regeneration after irradiation using gamma knife radiosurgery and identified molecular mechanisms that are shared between angiogenesis and neurogenesis during functional recovery from brain injury.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3317.
Abstract 3323: Induction of apoptosis in glioma cells by downregulation of X-linked inhibitor of apoptosis protein (XIAP)
Authors: Dasari Venkata Ramesh , Kaur Kiranpreet , Fassett Daniel , Klopfenstein Jeffrey D. , Dinh Dzung H. , Gujrati Meena , Rao Jasti S.
Published: February 2011

XIAP (X-linked inhibitor of apoptosis protein) is one of the most important members of the family of apoptosis inhibitors. XIAP blocks apoptosis by inhibiting both the initiator (caspase 9) and effector (caspase 3) caspases, and therefore, prevents cell death induced by various triggers. One mechanism through which tumor cells are believed to acquire resistance to apoptosis is by overexpression of XIAP. When XIAP is overexpressed, cancer cells are rendered resistant to apoptosis, both intrinsically and in response to chemotherapy and radiotherapy. It is upregulated in various malignancies, including human glioblastoma. It promotes invasion, metastasis, growth, and survival of malignant cells. In order to downregulate XIAP in glioma cells, we used human umbilical cord blood mesenchymal stem cells (hUCB) and siRNA of XIAP (siXIAP). We observed the effect of both hUCB and siXIAP on two malignant glioma cell lines (SNB19 and U251) and two glioma xenograft cell lines (4910 and 5310). In either case, proliferation of the glioma cells was significantly inhibited. Both treatments increased cytotoxicity of glioma cells and led to cell death. However, cytotoxicity is more significant in siXIAP treatments compared to hUCB cocultures. Both hUCB and siXIAP induced TNF-alpha-mediated apoptosis in glioma cells, which was confirmed by TUNEL assay, FACS analyses and immunoblotting. Apoptosis is characterized by loss of mitochondrial membrane potential and upregulation of mitochondrial apoptotic protein BAD. Cell death of glioma cells is marked by downregulation of Akt and phospho-Akt molecules. Taken together, our results indicate the therapeutic potential of XIAP and hUCB in treating malignant gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3323.
Abstract 3326: BEZ235 enhances radiosensitization via blockade of PI3-kinase/mTOR pathway in cisplatin-selected non-small cell lung cancer stem cells
Authors: Kim Kwang Woon , Jung Dae Kwang , Lu Bo
Published: February 2011

Purpose: Most drug resistance cancer cells have resisted to radiation therapy. In this study, we hypothesized that a dual inhibitor of PI3-kinase/ the downstream mammalian target of rapamycin (mTOR), BEZ235, could potentially enhance radiosensitization in cisplatin-selected non- small cell lung cancer stem cells.

Methods: Clonogenic assay was used to determine radiosensitivity. Stem cell marker was detected with CD133 antibody. P-AKT, P-mTOR and P-S6R were detected by Western Blotting. Cell cycle was performed using flow cytometry. Autophagy was determined by punctuate localization of GFP-LC3 fusion protein, which observed under a confocal fluorescence microscope in vitro.

Results: Basal CD133 which is a marker of stem cell, P-Akt, p-mTOR and P-S6R proteins were enhanced in cisplatin-selected non- small cell lung cancer stem cells (CSNSCLCSTC) when compared to parental non-small cell lung cancer (H460) cells. Cisplatin-selected non- small cell lung cancer stem cells mediate less radiation sensitivity when compared with parental cells in clonogenic assay(DER=0.82, p=0.02). However BEZ235 obviously enhanced sensitivity of cisplatin-selected non- small cell lung cancer stem cells to radiation (DER=1.2, p=0.01). Therefore, this enhanced radiosensitization was associated with a downregulation of PI3-kinase/mTOR signaling pathway and an increase in characteristic punctuate localization of GFP-LC3 as a marker for autophagy, suggesting that G2/M inhibition and autophagy were induced simultaneously.

Conclusions: These findings may be utilized as a novel strategy to enhance radiation therapy in drug-selected non-small cell lung cancer stem cells showing radioresistance.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3326.
Abstract 3329: Human prolactin increased chemosensitivity of breast cancer stem cells in MCF-7 and primary breast tumors from HER2/neu transgenic mice
Authors: Lee Eric , Park Jang-pyo , Chen Wen Y.
Published: February 2011

Background. Tumors are organized in a cellular hierarchy, in which cancer stem cells (CSCs) are the only cells that have the ability of self-renewal and differentiation. CSCs generally are in quiescent state and resistant to chemotherapy, which is one of the reasons that using conventional chemotherapeutics often leads to enrichment in CSCs. It is known that adult stem cells (ASCs) and CSCs share many common features. In adult mammary tissue, one of the distinctive features is its ability to go through multiple proliferation and involution cycles during pregnancy, suggesting that ASCs in this organ are readily responsive to female hormone regulations. One such hormone that plays a crucial role in this process is human prolactin (hPRL), which is required for mammary lobuloalveolar formation, terminal differentiation, and for synthesis of milk components. In this study, we hypothesize that simultaneous administration of hPRL with chemotherapeutic agents such as doxorubicin (Dox) will increase the cytotoxic effects of the chemotherapeutic drugs towards CSCs through the induction of CSCs proliferation/differentiation by hPRL.

Methods. MCF-7 human breast cancer cells and primary mouse breast cancer cells isolated from MMTV-neu transgenic mice were used in this study. Tumorsphere formation assay and ALDH activity measured by flow cytometry were used to measure the functional response of the CSCs after co-treatment of hPRL and Dox.

Results. (1) hPRL treatment increased tumorspheres formation in both MCF-7 (1.3-fold ± 0.24, p<0.05) and primary mouse breast cancer cells (2.3-fold ± 0.34, p<0.001). (2) Dox treatment (25ng/ml) increased ALDH+ cell population (7.74-fold ± 0.42, p<0.001), despite the fact that there is no significant change in total tumorspheres number (3) Combination of hPRL and Dox treatment decreased 50% of tumorspheres formation compared that of Dox treatment alone.

Conclusion. Our preliminary results suggest that hPRL is able to improve Dox's efficacy towards CSCs. Further studies are needed to validate the results. This novel therapeutic approach may open doors for designing new strategy for breast cancer treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3329.
Abstract 3336: Upregulated inflammatory cytokines in glioma stem cells: Targets for glioma therapies
Authors: Wang Lei , Real Dalila Del , Seo Gwi-Moon , Pyle Marla , Troyer Dery
Published: February 2011

Gliomas are the most common primary brain tumors in adults. With the highly invasive growth pattern, gliomas often escape current therapeutic modalities including combinations of surgery, radiotherapy and chemotherapy. Recent studies implicate that a minority population of glioma stem cells (GSCs) are responsible for glioma maintenance and recurrence. Interleukin (IL)-1 beta, IL-6 and IL-8 are pleiotropic proinflammatory cytokines and their excessive production in the tumor microenvironment is associated with poor disease outcome in many cancer types, including glioma. These cytokines promote tumor growth via induction of proteins associated with tumor invasion and angiogenesis and anti-apoptosis. We hypothesize that GSCs acquire their chemo-resistance and invasion by excessive cytokine production. To test the hypothesis, we compared the cytokine gene expression of a human GSC-enriched population in serum-free medium (SFM) in the presence of epidermal growth factor and fibroblast growth factor, to that of glioma cells in serum-containing medium (SCM). SFM is the neural stem/progenitor cell (NPC) medium in which NPCs proliferate in neurospheres and this medium also has been used to enrich GSCs. We found that IL-1 beta and IL-6 were upregulated in GSC-enriched population along with increased expression of Nestin, a marker of neural stem cells. More interestingly, Smad interacting protein 1 (SIP1), which is thought to be involved in glioma cell migration and invasion, was also increased in the GSC-enriched population. The paralleled upregulation of IL-1 beta and IL-6 with Nestin and SIP1 in GSC-enriched population suggest that there is a possible correlation between them. The correlation is now under investigation. Since IL-1 beta is elevated in GSC enriched population, we further determined whether IL-1 beta plays a critical role in glioma growth. Glioma cells were co-cultured with mouse NPCs expressing IL-1 receptor antagonist (IL-1ra), which can bind to IL-1 receptors without transmitting activation signals and represents a competitive inhibitor of IL-1. Proliferation of glioma cells, which were modified to express Renilla luciferase, was measured using bioluminescence imaging. Our results showed that proliferation of glioma cells was inhibited in the co-culture with NPCs expressing IL-1ra, but not in the co-culture with NPCs without expression of IL-1ra, compared to glioma cells cultured alone. The results suggest that IL-1 has an important role in glioma cell growth and NPCs engineered to express IL-1ra have therapeutic potential for gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3336.