bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2222: Mutational and copy number analysis of the PI3K/Akt/mTOR pathway in oral cavity squamous cell carcinoma
Authors: Morris Luc GT , Ganly Ian , Heguy Adriana , Chan Timothy A.
Published: February 2011

The phosphatidylinositol 3-kinase signaling (PI3K) pathway is integral to cell growth, proliferation and survival, and is upregulated in multiple human cancers. Several oncogenes and tumor suppressors within this pathway are altered by mutation, amplification or deletion, and are currently being investigated as therapeutic targets. The pattern of dysregulation in this pathway differs among various cancers, and has not been comprehensively characterized in oral squamous cell carcinoma.

After institutional review board approval, fresh frozen tumor and matched normal tissues were obtained from 32 patients treated for oral cavity squamous cell carcinoma, and genomic DNA was extracted. Using high-throughput Sanger sequencing, the 23 component genes of the PI3K/Akt/mTOR pathway were comprehensively sequenced. Putative mutations identified by mutation detection software were validated on a second PCR and sequencing procedure. Array CGH was performed on the Agilent 1M microarray, and data analysis performed using the RAE method.

Component genes within the PI3K/Akt/mTOR pathway were comprehensively sequenced, including AKT1, AKT2, AKT3, AKTS1, AXL, AGAP2, MTOR, GBL, GOLPH3, RAPTOR, MUC1, PDPK1, PIK3CA, PIK3CB, PIK3CD, PIK3CG, PIK3R1, PIK3R2, PTEN, RHEB, RICTOR, TSC1 and TSC2. Activating mutations were identified in PIK3CA in 2 of 32 (6.3%) specimens. No other genes harbored somatic mutations. DNA copy number gains and losses are described in detail.

In the first comprehensive mutational and copy number analysis of 23 component genes of the PI3K/Akt/mTOR pathway in cancer, we report a low prevalence of somatic mutations, and a higher frequency of copy number alteration in oral cancer specimens. In contrast to colorectal, breast, ovarian, brain and bladder cancers, mutation is not the primary source of activation of this pathway in oral 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 2222.
Abstract 2223: Screening for oncoviruses using next-generation sequencing
Authors: El-Dinali Mohamed , Braegelmann Johannes , Salgia Ravi , Seiwert Tanguy
Published: February 2011

Introduction: During the last decades numerous viruses have been found to play a causative role in cancer. Besides well established connections like HPV in cancers of the uterine cervix or EBV in nasopharyngeal carcinoma recent advances in next-generation sequencing technologies have led to the discovery of new tumor viruses e.g. a previously unknown Polyoma-virus subtype linked to Merkel-cell carcinoma of the skin.

Method: To gain insight into the mechanisms leading to tumor formation we sequenced the transcriptome from an HPV16 positive cell line using short-read Illumina RNA-Seq and established a computational pipeline to screen the short reads for viral sequences based on freely available alignment tools.

Results: Due to the large amount of data and in order to narrow down the search space and reduce the chance for false positives the first screening step consists in a subtraction of sequences that align to the human reference genome reducing the amount of reads by ∼70%. To find both known viruses and yet unknown strains/subtypes we apply a two-fold approach:

1. A rapid search for known viruses is done using a short-read alignment tool with a set of stringent parameters to map non-human reads to an index of the viral genbank and subsequent quantification of the hits.

2. Since slight differences between read and index sequence will cause the rapid search to not call a match we implemented a more lenient approach using standalone BLAST.

Applying approach 1) to the control cell line we clearly identified HPV16 as being present. To test the possibility of finding a potentially unknown virus we deleted HPV16 from both the rapid approach and the index which caused the first algorithm to no longer identify the virus. Running the BLAST basec approach 2) resulted in multiple hits the most prominent being alignments to HPV subtypes 35 and 58, thus indicating the successful identification of related strains.

Conclusion: We demonstrated the feasibility of our dual viral screening approach in an in-silico experiment using an HPV16 positive control. Our approach appears to be able to identify novel related viruses. In the future this will allow screening of patient samples and viral screening was added to our data pipeline.

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 2223.
Abstract 2236: Nuclear roundness variance predicts prostate cancer progression, metastasis and death: A prospective evaluation with up to 25 years of follow-up after radical prostatectomy
Authors: Veltri Robert W. , Isharwal Sumit , Miller Craig , Epstein Jonathan I. , Partin Alan W.
Published: February 2011

Introduction and Objective: Nuclear structure is often altered in cancer due to spatial rearrangements of chromatin organization via activation of oncogenes and other chromatin remodeling genes. Therefore, we evaluated the prognostic value of nuclear roundness variance (NRV) for prostate cancer (PCa) progression, metastasis and PCa-specific death-free survivals in a cohort of 116 men after radical prostatectomy (RP).

Methods: For each patient, ∼150 intact nuclei from the primary tumor were captured using the DynaCELL Motility Morphometry Measurement workstation (JAW Associates, Inc., Annapolis, MD) at a magnification of 1250X in 1992-93. Next, nuclear roundness variance (NRV) was calculated for each of the 116 cases using the this unique shape factor from ∼150 nuclei captured. The resulting variance values were multiplied by 10,000 to determine the NRV factor for each case. NRV data was merged with clinical, pathologic and follow-up data for all patients in 2009. Cox proportional hazards regression was used to identify significant prognostic factors for prediction of progression, metastasis and PCa-specific death free survivals.

Results: Median follow-up time after RP for all patients was 19.0 years (range: 1-25 years, mean: 17.43 years), with approximately 92% (107/116), 71% (82/116) and 47% (55/116) patients having ≥10 years, 15 years and 20 years of follow-up respectively. NRV was the most significant parameter for prediction of all three outcomes and its concordance-index (C-Index) increased from progression (0.7080) to metastasis (0.7332) to PCa-specific death (0.8090) free survival predictions. Of note, NRV C-Index was significantly higher compared to Gleason Score C-Index for metastasis (0.7332 vs. 0.6046; p = 0.027) and PCa-specific death (0.8090 vs. 0.6336; p = 0.004) free survival predictions. However the difference between NRV and Gleason Score C-Indexes was not statistically significant for progression free survival prediction (0.7080 vs. 0.6463; p = 0.106).

Conclusions: Variance of nuclear roundness (a measure of tumor heterogeneity) is a valuable nuclear structural feature to assess an aggressive phenotype of prostate cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2236.
Abstract 2240: Clinical significance of CEA, PRL3 and EZH2 transcripts expressed in lymph nodes of colorectal cancer patients
Authors: Lagarde Alain E. , Wood Caroline , Guindi Maha , Stern Hartley S.
Published: February 2011

Objective: The role of micrometastases detected immunologically and molecularly in lymph nodes of colorectal cancer patients remains elusive. We investigated the prognostic significance of CEA, PRL3 and EZH2 expressed in lymph nodes of 42 pathologically node-negative (pN0) colorectal patients and 13 stage III/IV patients as controls. Seventeen pN0 patients (40%) received adjuvant therapy after curative surgery.

Experimental design: CEA, PRL3 and EZH2 transcripts were amplified by reverse transcription PCR in nearly 600 paraffin embedded lymph nodes. On average 9.7 ± 5.7 nodes were harvested per case. After 55 months median follow-up, gene expression was correlated with recurrence-free (RFS), overall (OS) and disease-free survival (DFS), using Kaplan-Meier analysis and log-rank test for comparison.

Results: The proportion of cases demonstrating CEA+, PRL3+ and EZH2+lymph node expression increased from stage I to III/IV and reached 71%, 76% and 26% respectively in the pN0 group. EZH2 had no significant prognostic value among pN0 patients. Disease progressed more rapidly (50% DFS at 30 versus 53 months) in EZH2+ than EZH2- node-positive patients. CEA+ was associated with ∼18% lower RFS/OS, and 8% lower DFS than CEA- among therapeutically-untreated patients, those with colon and with stage II tumors. pN0 patients with PRL3+ nodes had better outcomes than PRL3- cases (DFS, P= 0.059), namely in the colorectal (OS, P= 0.056; DFS, P= 0.12), colon (DFS, P= 0.056) and stage II tumor in the non-adjuvant settings (DFS, P= 0.0047).

Conclusions: Lymph node expression of the PRL3 phosphatase gene predicted more favorable outcomes for pN0 colorectal cancer patients. The role of CEA as an adverse predictor of survival was confirmed albeit less significant than previously reported. Neither CEA nor PRL3 nodal expression was predictive of rectal cancer recurrence.

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 2240.
Abstract 2243: Prognostic significance of nuclear Sox4 expression in cutaneous melanoma and its role in cell migration
Authors: Shourkaei Seyed Mehdi Jafarnejad , Wani Aijaz Ahmed , Martinka Magdalena , Li Gang
Published: February 2011

Sry-related high-mobility group (Sox) family of genes is consisted of transcription factors involved in various physiological processes including embryonic development, cell fate decision and differentiation. Deregulated expression of Sox4 has been reported in several human cancer types so far; nevertheless the expression and possible role of Sox4 in human melanoma cancer progression is yet to be identified. To investigate the putative role of Sox4 in the development of melanoma and its correlation with patient survival, we examined the expression of Sox4 protein in different stages of melanocytic lesions and its role in melanoma cancer cells proliferation and migration. Using tissue microarray and immunohistochemistry, we evaluated Sox4 protein expression in 43 dysplastic nevi, 89 primary melanomas, and 48 metastatic melanomas. We observed a marked reduction in nuclear Sox4 expression in malignant melanomas compared with dysplastic nevi (Kruskal-Wallis test; p < 0.05) and primary melanomas (Kruskal-Wallis test; p < 0.01). Moreover, the reduced nuclear Sox4 expression was significantly correlated with a poorer disease-specific 5-year survival of patients with primary and metastatic melanoma (Kaplan-Meier test; P = 0.039). Multivariate Cox regression analysis revealed that reduced nuclear Sox4 expression is an independent prognostic factor to predict patient outcome (p = 0.049). We also knocked down the expression of Sox4 in MMRU human melanoma cell line by siRNA and the migratory ability of the cells as well as their proliferation rate were analyzed following Sox4 silencing. Strikingly, the wound healing assay revealed an augmented migration ability of MMRU cells upon siRNA mediated knockdown of Sox4. However, the absence of Sox4 did not affect the rate of melanoma cancer cells proliferation. Our results suggest that Sox4 may be a suitable marker for the prognosis of the melanoma cancer outcome and a potential therapeutic target for human melanoma.

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 2243.
Abstract 2244: Internalization of basigin-2, a poor prognosis marker, by a monoclonal antibody
Authors: Lappi Douglas A. , Ancheta Leonardo , Nowak Romana A.
Published: February 2011

Basigin-2 (EMMPRIN, CD147) is widely expressed on the surface of numerous tumor types and is associated with poor prognosis. Basigin is an integral membrane glycoprotein also expressed on numerous normal cell types. Several functions have been described for basigin-2 within both normal and malignant tissues. The best characterized is its ability to induce the expression of matrix metalloproteinases in stromal cells. As such, it is suggested to play a role in tumor metastasis. Because of its association with poor prognosis and possible role in metastasis, we examined the ability of basigin-2 to be targeted on the cell surface and internalized as a possible gateway for entry into tumor cells, or for down-regulation for inhibition of metastasis. Belton et al. have documented internalization of basigin-2 by a recombinant soluble protein corresponding to the extracellular domain (ECD) of basigin-2 (J Biol Chem, 2008, 283:17805).

We have extended this investigation to examine antibody-mediated endocytosis. A monoclonal antibody to the basigin ECD (also reported in Belton et al.) has been used in an internalization assay using biotinylated antibody mixed with streptavidinylated-saporin (the ribosome-inactivating protein). This complex will eliminate cells only if there is antibody-mediated endocytosis, since saporin has no mechanism of cell entry. HeLa cells, which are known to express basigin-2, were challenged and a potent cytotoxicity was observed: the ED50 of the biotinylated antibody-streptavidin-saporin complex is 370 pM. This is evidence that the antibody internalizes in HeLa cells. Similar data were observed with ovarian carcinoma cells (OVCAR3, SKOV3). Non-tumor cells with low expression have low sensitivity to the complex and there was a correlation between basigin-2 expression on the cell surface and cytotoxicity.

We conclude that basigin-2 internalizes in response to antibody binding. Further work is necessary to determine if this can be used for introduction of molecules into basigin-2-expressing cells or if down-regulation would have an effect on metastasis promoted by this poor prognosis molecule.

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 2244.
Abstract 2250: Aberrant leukocyte infiltration as a trigger for tumor metastasis
Authors: Hsiao Yi-Hsuan , Man Yan-Gao
Published: February 2011

Our previous studies have led to a novel hypothesis that aberrant leukocyte infiltration is a trigger for tumor invasion. Our hypothesis and supporting data have been published in multiple peer-reviewed journals. Our current studies attempted to further expand our previous observations, and to test a novel hypothesis that aberrant leukocyte infiltration may also represent a trigger for tumor metastasis. Tissue sections from human breast, prostate, cervical, and lung tumors were analyzed with multidisciplinary approaches with a panel of tumor metastasis-related biomarkers. Our study revealed that pre-invasive tumors surrounded by or immediately adjacent to leukocyte aggregates displayed a number of unique features: [1] significantly reduced expression of cell surface adhesion molecules, [2] significantly elevated blood vessels and lymphatic ducts at the intersection between the tumor core and leukocyte aggregates, [3] the presence of isolated individual tumor cells or cell clusters within the leukocyte aggregates, [4] physical attachment or fusion of tumor cell membrane and condensed chromosomes of mitotic tumor cells with leukocytes, and [5] the presence of co-joined tumor cells and leukocytes within vascular structures or lymphatic ducts. Together, our findings suggest that leukocyte infiltration may trigger tumor metastasis through three correlated mechanisms: (1) to physically disrupt the inter-cellular junctions and cell surface adhesion molecules, [2] to interfere with the mitotic process or physically drag tumor cells to different sites during their migration, and [3] to stimulate growth of vascular structures that facilitate tumor metastasis.

(Supported in part by grants DAMD17-01-1-0129, DAMD17-01-1-0130, PC051308 from Congressionally Directed Medical Research Programs, grant BCTR0706983 from The Susan G. Komen Breast Cancer Foundation, and grant 2008-02 from The US Military Cancer Institute and Henry M. Jackson Foundation to Dr. Yan-gao Man, and by grant 2006CB910505 from The Ministry of Chinese Science and Technology Department to Drs. Xichen Zhang and Yan-gao Man).

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 2250.
Abstract 2253: Proteases and cytokines in the transition from preinvasive DCIS to invasive breast carcinoma
Authors: Sameni Mansoureh , Anbalagan Arulselvi , Sloane Bonnie F.
Published: February 2011

The transition from preinvasive ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) involves alterations in proteolytic pathways of both epithelial and stromal cells. Infiltration of macrophages, angiogenesis and secretion of chemotactic molecules by DCIS and surrounding stromal cells also facilitate this transition. We have developed a 3D coculture model, MAME (mammary architecture and microenvironment engineering), to study the role of proteases and cytokines in the progression from pre-invasive DCIS to IDC. MAME-A recapitulates the architecture of normal human breast tissue with the epithelial and stromal components growing in overlays of basement membrane and stromal collagen, respectively. In MAME-B, the stromal collagen is eliminated.

The cells used in our studies were human MCF10.DCIS cells, WS-12Ti human breast tumor-associated fibroblasts, U937 human monocytes and human umbilical vein endothelial cells (HUVEC). We analyzed proteolysis through live cell imaging of the degradation of quenched fluorescent matrix proteins [DQ-collagen I™ mixed with collagen I and DQ-collagen IV™ mixed with reconstituted basement membrane (rBM)].

DCIS cells in MAME-A cocultures with fibroblasts exhibited invasiveness at 24 days, yet at only 16 days in MAME-B cocultures. Expression and secretion of cytokines, especially IL-6, were elevated in these cocultures. The invasiveness of the DCIS cells in the cocultures with fibroblasts could be reduced by addition of neutralizing antibody to IL-6. Addition of either monocytes or endothelial cells reduced the time at which invasiveness occurred to as early as 4 days in MAME-B cocultures. Addition of monocytes increased secretion of monocyte chemoattractant protein-1 (MCP-1), IL-8 and GRO. The combination of a neutralizing antibody to hMCP-1 and a receptor-blocking antibody to hCXCR2 (a receptor for IL-8 and GRO) reduced invasiveness more effectively than did either of the antibodies used alone. When HUVEC were grown in MAME-B cocultures with DCIS or DCIS and fibroblasts, the HUVEC grew into a network of tubules with distinct lumens over a period of 4 days. In 3D and 4D reconstructions of these same cocultures, DCIS cells could be seen to migrate toward the HUVEC tubules and invade into their lumens. These phenomena occurred in parallel with increases in proteolysis and in expression and secretion of the cytokines MCP-1, IL-8, RANTES and GRO.

The MAME models provide a system for the study of proteolytic and cytokine pathways and the interactions between these pathways that are associated with the transition from DCIS to invasive ductal carcinoma. This model could be used as an in vitro tool for screening drugs that target these pathways.

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 2253.
Abstract 226: Aspirin alone or in combination with doxorubicin induced efficient apoptosis in human hepatocellular carcinoma cells
Authors: Hossain Mohammad Akbar , Kim Min Young , Kim Dong Hwan , Kim Jin-Ah , Kang Yong Jung , Moon Jeon Ok , Kim Gi Young , Choi Yung Hyun , Kim Nam Deuk
Published: February 2011

Hepatocellular carcinoma (HCC), one of the most common cancers worldwide, is refractory to most anticancer drugs. Doxorubicin offers a survival benefit to patients with intermediate hepatocellular carcinoma. However, the favorable therapeutic response to doxorubicin is often associated with severe toxicity. The present research was aimed at developing a strategy of increasing doxorubicin sensitivity so that lower doses may be used without compromising efficacy. Recently, aspirin (ASA) and some other nonsteroidal anti-inflammatory drugs (NSAIDs) have drawn much attention for their protective effects against colon cancer and cardiovascular disease. Several studies also found that regular ASA use is significantly associated with reduced risk of lung cancer. We investigated the effects of ASA on HepG2 human hepatocellular carcinoma cell line. Growth inhibition effect of aspirin was measured by MTT assay. Flow cytometry analysis revealed that aspirin can induce G2/M phase arrest which we observed by the level of cell cycle regulatory proteins and DNA ladder formation indicates apoptosis dose dependently. We found that ASA treatment followed both extrinsic and intrinsic pathways for apoptosis induction. In addition to this, when administered in combination, doxorubicin (DOX) and ASA produced synergistic effect in growth inhibition in human hepatocellular carcinoma cells and induced apoptosis in caspase-dependent pathway. Therefore, clinically achievable concentrations of DOX and ASA used in combination may produce a strong anticancer synergy that warrants investigation as a therapeutic strategy to enhance the efficacy of doxorubicin for treating HCC. [This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korea government (MOST) (R01-2006-000-11117-0)].

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 226.
Abstract 2280: Endothelin axis in bladder cancer metastasis
Authors: Said Neveen A. , Theodorescu Dan
Published: February 2011

In bladder cancer, recurrence after surgical intervention for muscle invasive disease is problematic as nearly half of patients harbor occult distant metastases, mainly in the lungs, with poor 5-year survival rate. Our prior work has indicated that RhoGDI2 reduces tumor Endothelin 1 (ET-1) secretion and this may contribute to its metastasis suppressor function, however how the endothelin axis acts to promote lung metastasis is unclear. We have found from gene expression profiling of human bladder tumors that high ET-1 expression was correlated with poor survival. Here we also identify ET-1 as a novel mediator of bidirectional cross talk between tumor cells and inflammatory macrophages. Tumor-derived endothelin, through its cognate endothelin receptor A (ETAR) exerts autocrine and paracrine effects on tumor cells and macrophages, significantly inducing its own production, by both cell types, concomitant with triggering an early inflammatory response in the lungs, which progressively increases till development of lung metastases. Abrogation of ET-1/ETAR axis by knock down tumor cell ET-1 by short hairpin RNA (shRNA) or systemic administration of ETAR blocker (ZD4054) significantly suppressed the development and the progression of this lung inflammatory response, and decreased the development of microscopic and macroscopic lung metastases, together with improvement of animal survival. In vitro, genetic manipulation of ET-1, ETAR and ETBR in bladder cancer cell lines co-cultured with macrophages revealed that ET-1 augments the inflammatory response through activation of transcriptional factors NFκB and AP-1, key orchestrators of inflammation, in both cancer cells and macrophages. The reciprocal activation of both transcriptional factors was mediated by ET-1/ETAR axis in cancer cells, whereas in macrophages it was mediated by ET-1/ETAR and ETBR axis. These findings highlight a molecular mechanism that can be pharmacologically manipulated to enhance the efficacy of current adjuvant approaches to muscle invasive bladder cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2280.
Abstract 2283: The role of insulin-like growth factor-1 and insulin like growth factor-1 receptor in obesity and oesophageal cancer
Authors: Doyle Suzanne L. , Donohoe Claire , Lysaght Joanne , Lithander Fiona , Pidgeon Graham , Reynolds John V.
Published: February 2011

Introduction: The insulin-like growth factor pathway, which is involved in cell proliferation, differentiation and protection against apoptosis, has been implicated in esophageal cancer progression. This study examined the role of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-1 receptor (IGF-1R) as a potential mechanism linking obesity with esophageal cancer.

Methods: IGF-1R expression in esophageal squamous cell carcinoma (KYSE, OE21) and adenocarcinoma (OE19, OE33) cell lines was measured by Western analysis and the effect of IGF-1 (0.1 − 1000 ng/ml) on survival of cell lines in relation to their expression of IGF-1R was assessed by MTT assay. Circulating levels of IGF-I were measured in 50 human serum samples by ELISA and immunohistochemistry determined IGF-1R expression in human tumor sections. Results were related to tumor type, BMI and survival.

Results: Higher IGF-1R expression was observed in squamous cell carcinoma cell lines, however only adenocarcinoma cell lines showed significantly increased cell survival in response to IGF-1 (p <0.001). Significantly higher circulating levels of IGF-1 were observed in patients with adenocarcinoma (p = 0.02). In resected esophageal tumor sections, no statistical difference between intensity of IGF-1R expression and BMI or survival was observed, however reduced survival was observed in patients with moderate-strong compared to weak-negative IGF-1R expression. Higher intensity IGF-1R staining was observed in the ‘immune cell rich’ leading edge of adenocarcinomas.

Conclusion: This study suggests an important role for IGF-1 in esophageal adenocarcinoma cell survival. Novel findings relating to high IGF-1R expression in the leading edge of esophageal adenocarcinoma and possible associations with obesity are also presented.

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 2283.
Abstract 2292: Loss of hCLCA2 promotes EMT in breast cancer
Authors: Elble Randolph C. , Walia Vijay
Published: February 2011

Epithelial cells acquire mesenchymal characteristics by undergoing phenotypic and genotypic changes during cancer progression. An early step in the epithelial to mesenchymal transition (EMT) is the disruption of intercellular connections due to loss of epithelial cadherins. We find that expression of tumor suppressor hCLCA2 is strongly associated with epithelial differentiation and that induction of EMT by mesenchymal transcription factors represses its expression. Moreover, we report here that knockdown of hCLCA2 by RNA interference results in disruption of cell-cell junctions by downregulating E-cadherin. This also imparts invasiveness and anoikis-resistance to epithelial cells but is insufficient to induce full EMT. However, activation of Ras oncogene in combination with hCLCA2 knockdown is sufficient to induce full EMT in vitro. These findings indicate that, like E-cadherin, hCLCA2 is required for epithelial differentiation and suggest that its loss during tumor progression contributes to metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2292.
Abstract 2296: Sorafenib inhibits hepatocyte growth factor-induced epithelial mesenchymal transition in hepatocellular carcinoma
Authors: tomoyuki nagai , Arao Tokuzo , Furuta Kazuyuki , Sakai Kazuko , Kudo Kanae , Kaneda Hiroyasu , Tamura Daisuke , Aomatsu Keiichi , Fujita Yoshihiko , Matsumoto Kazuko , Saijo Nagahiro , Kudo Masatoshi , Nishio Kazuto
Published: February 2011

Sorafenib is a multikinase inhibitor with activity against Raf kinase and several receptor tyrosine kinases, including vascular endothelial growth factor receptor2, platelet-derived growth factor receptor. Epithelial mesenchymal transition (EMT) describes a process wherein static epithelial cells lose cell-cell contacts, acquire mesenchymal features and manifest a migratory phenotype, mediating malignant phenotypes for cancer cells. It remains unclear for involvement of Sorafenib against EMT, especially in hepatocellular carcinoma (HCC).

Hepatocyte growth factor (HGF) activated HGF/MET signaling pathway in HepG2 and Huh7 cells and resulted in cell scattering and spindle-shaped morphology, changes that are characteristic of EMT. SNAI1 is a key transcription factor known to be involved in EMT. HGF up-regulated SNAI1 and N-cadherin expression, and down-regulated E-cadherin in a dose- and time-dependent manner. Knockdown of SNAI1 by siRNA almost completely inhibited these morphological changes, indicating that SNAI1 was required for HGF-induced EMT in HCC cell lines. Similarly, Sorafenib and the MEK inhibitor U0126 markedly inhibited the HGF-induced SNAI1 expression and these morphological changes at 10 μM of concentration, whereas PI3 kinase inhibitor Wortmannin did not.

In conclusion, Sorafenib exerts a potent inhibitory activity against EMT via ERK/SNAI1 in HCC. This activity may provide an additional clinical benefit in patients with HCC.

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 2296.
Abstract 2312: High ΔN/TA TP63 isoforms ratio is liked to the maintenance of an epithelial phenotype in Squamous cell lung cancer
Authors: Pallier Karine , Desroches Marine , Leschi Cristina , Khattabi Leila El , Cazes Aurelie , Danel Claire , Riquet Marc , Laurent-Puig Pierre , Blons Helene
Published: February 2011

Squamous cell carcinoma of the lung (SCC) is a tumor that arises from bronchial epithelium. The transcription factor TP63, member of the TP53 family is crucial for epithelial development and different isoforms can be reactivated during carcinogenesis. Its role remains controversial and its relation with acquired invasive phenotype and epithelial to mesenchymal transition has not been fully explored in lung cancer. Here we report the phenotypic characterization of a series of 41 lung SCC, collected according to French Laws (CPP Ile de France II, 2008-A01074-51) and for which frozen tumor tissues were available for RNA extraction.

Epithelial and mesenchymal markers: CDH1 (E-cadherin), JUP (Plakoglobin), VIM (Vimentin), CDH2 (N-cadherin) and TP63 isoforms (ΔNTP63 and TATP63) as well as TWIST1 and SNAIL reported to be linked to EMT and to interact with TP63 were analyzed at RNA level. Moreover, TP63 amplification was determined by quantitative PCR. We used real RTPCR to estimated the expression of each marker, a pool of non tumor lung tissue RNA (6 individuals) was used as reference as well as 3 reference genes (RPL13, YAP1, POLR2A) to calculate the ΔΔCt. CDH1, JUP, VIM, CDH2, ΔN and the TA TP63 isoforms were analyzed using SYBR Green technology; TWIST and SNAIL were analyzed using TaqMan probes with similar references.

In this series, epithelial markers were associated together (p=0.001) as well as mesenchymal markers (p=0.0003). We confirmed that the TP63 locus at 3q28 was commonly amplified in lung SCC but this copy number change was not related to high ΔN/TA ratio (p=0.23). High ΔN/TA ratio was linked to high CDH1 expression (p=0.04), high JUP expression (p=0.01) or both (p=0.02) but not to high VIM or CDH2. At the opposite, TWIST1 expression was related to high VIM (p=0.02) and high CDH2 (p=0.008) or both (p=0.01). SNAIL was not expressed in this tumor series. TP53 mutations were not related to TP63 amplification nor to high ΔN/TA ratio and both event seem to be independent in this series.

To conclude, the expression of epithelial and mesenchymal markers did not match to the high, moderate or low differentiation status determined by the pathologist and allowed the clustering of different tumor groups. In this tumor series, high ΔN/TA ratio was linked to the maintenance of epithelial characteristics whereas low ΔN/TA ratio and TWIST expression was related to EMT. Clinical data analysis is ongoing to study the impact of these associations on patient outcome.

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 2312.
Abstract 2314: Epithelial-mesenchymal transition derived cells exhibit multi-lineage differentiation potential similar to mesenchymal stem cells
Authors: Battula Venkata Lokesh , Evans Kurt , Hollier Brett G. , Shi Yuexi , Marini Frank C. , Ayyanan Ayyakkannu , Brisken Cathrin , Guerra Rudy , Andreeff Michael , Mani Sendurai A.
Published: February 2011

The epithelial-to-mesenchymal transition (EMT) is an embryonic process that becomes latent in most normal adult tissues. Recently, we have shown that induction of EMT endows stem cell traits to breast epithelial cells. Mesenchymal stem cells (MSC) have the capacity to self-renew and differentiate into multiple tissue lineages. We hypothesized that the activation of EMT by ectopic expression of Twist, Snail or TGF-β in immortalized human mammary epithelial cells (HMEC) will result in the generation of cells with a phenotype and functionality similar to MSC. We found that the EMT-derived cells not only showed similar morphology but also displayed the typical MSC phenotype i.e. CD44+, CD24− and CD45−. Alternatively, MSC expressed EMT inducing genes such as Twist, Snail and FOXC2. Interestingly, CD140b (PDGFR-β), a marker for naive MSC, was exclusively expressed in EMT-derived cells compared to their epithelial counterparts. Moreover, functional analysis revealed that EMT-derived but not the control cells differentiate into Alizarin Red S-positive mature osteoblasts, Oil Red O-positive adipocytes and Alcian Blue-positive chondrocytes similar to MSC. We also observed that EMT-derived but not control cells invade and migrate towards MDA-MB-231 tumor cells in-vitro similar to MSC, displaying the characteristic tropism of MSC for tumor cells as previously reported by us. In-vivo wound homing assays in nude mice revealed that the EMT-derived cells home to wound sites similar to MSC. In conclusion, we demonstrated that the EMT-derived cells are similar to MSC in gene-expression, multi-lineage differentiation, migration towards tumor cells and their ability to home to wounds. These results also suggest that EMT-derived MSC are active participants in cancer growth and 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 2314.
Abstract 2316: Epithelial to mesenchymal transition: Interplay between E-Cadherin and HGF/SF-Met signaling
Authors: Gao Chong-Feng , Haak Elizabeth A. , Su Yanli , Hudson Eric , Dykema Karl , Vande Woude George F. , Vande Woude George F.
Published: February 2011

During malignant progression, primary tumor cells switch from proliferative to invasive phenotypes and switch again at low frequency to form proliferating metastases. The acquisition of the invasive phenotype through epithelial-mesenchymal-transition (E-M-T) is a critical step in tumor progression, as is the mesenchymal-epithelial-transition (M-E-T) reverting to a proliferative colony or metastases. To study the mechanism(s) underlying phenotypic switching, we isolated HGF/SF-inducible invasive mesenchymal (M) and non-invasive epithelial (E) subclones from prostate and ovarian carcinoma cell lines based on three dimensional branching morphogenesis assays. We show that invasive M subclones can be generated from originally homogeneous non-invasive E subclones and phenotype switching occurs with loss of E-Cadherin and concomitant gain of Met signaling. By contrast, tumor xenograph formed by invasive M subclones partially revert to epithelial phenotype with gain of E-Cadherin and loss of HGF/SF-Met signaling, suggesting an interplay between E-Cadherin and HGF/SF-Met signaling in regulating reversible phenotypic switching. We have further studied the mechanisms regulating phenotypic switching. Our results explicate how intrinsic alteration in tumor cell properties (E-M-T) and extrinsic signals from microenvironment (HGF/SF) interact in regulating tumor phenotypic switching.

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 2316.
Abstract 2320: N-cadherin down-regulation has antitumor effects in esophageal cancer
Authors: Nishitani Seishi , Noma Kazuhiro , ohara Toshiaki , tanabe shunsuke , fujiwara yasuhiro , Motoki Takayuki , sakurama Kazuhumi , Takaoka Munenori , Shirakawa Yasuhiro , Yamatuji Tomoki , Matuoka Junji , Naomoto Yoshio , Haisa Minoru
Published: February 2011

Tumor microenviroment plays a critical role for cancer and their malignancy. Dynamic changes in tumor microenvironment have been observed in many tumors including esophageal cancer. And during the drastic change, some of tumor cells show EMT (epithelial mesenchymal transition) which means that epithelial cells lose their epithelial feature and acquire mesenchymal properties, as invasiveness and also communication with other kinds of mesenchymal cells. During those change, one of the marker of epithelial feature, E-cadherin is switch to N-cadherin, it has been known as “cadherin switch”. And it is thought that those changes have relations with cancer malignancy.

Previously we reported that esophageal cancer cells secrete TGF-b1 and activate mesenchymal fibroblasts, and those activated fibroblasts guided more endothelial cells into tumor and induced angiogenesis. In the other hand, we hypothesis cell-cell direct communication like cell-cell contact is also important role for cancer progression.

So in this report, firstly we analyze the expression of cadherins in surgical specimens in immunohistochemistry and determined expression patterns of N-cadherin in esophageal squamous cell carcinoma and Barrett's adenocarinoma. Some of those cancers showed N-cadherin expression and specifically expressed in their invasive front. Secondary we analyzed cancer cell lines as well, TE-1, TE-4, TE-10, OE19 and OE33. Those cell lines are squamous cell carcinoma cell lines and Barrett's adenocarcinoma cell lines, and those showed expression of E-cadherin and some of them showed expression of N-cadherin. And we used those cell lines and explored the behavioral differentiations of each cell lines.

Here, we report and show analysis of E- or N- cadherin expression in surgical specimens and cancer cell lines. And also we could show the potential of regulation of cadherins in esophageal cancer as a new therapeutic target, which is well recognized as one of the most aggressive malignancies in the world.

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 2320.
Abstract 2340: Lactic acidosis triggers starvation response with distinct metabolic profiles by the induction of TXNIP through MondoA
Authors: Chen Julia, Ling-Yu , Merl Daniel , Peterson Chris , Wu Jianli , Liu Patrick Yantyng , West Mike , Ayer Don , Chi Jen-Tsan
Published: February 2011

Although lactic acidosis and other tumor microenvironmental stresses are prominent features in solid tumors, how they influence the phenotypes of cancer cells are not well understood. We compared the transcriptional response of breast cancer cells under three distinct tumor microenvironmental stresses – lactic acidosis, glucose deprivation and hypoxia. We found that lactic acidosis induced gene expression highly similar to glucose deprivation and triggered features of starvation response. In spite of their similar transcriptional response, lactic acidosis had opposite effects on the glucose uptake to glucose deprivation. This discrepancy guided the supervised analysis of expression data to identify TXNIP as the potential regulator for such divergent metabolic effects. TXNIP and its paralogue ARRDC4 were both induced under lactic acidosis and repressed with glucose deprivation. Their expression was also highly correlated with predicted lactic acidosis pathway activities and associated with favorable clinical outcomes in human cancers. These findings are due to the ability of TXNIP to inhibit the tumor glycolytic phenotypes and redirect energy generation toward aerobic respiration. The induction of TXNIP under lactic acidosis is caused by the activation of the glucose-sensing helix-loop-helix transcriptional complex MondoA: Mlx. Lactic acidosis activates the MondoA-TXNIP pathways and contributes to its “anti-Warburg” metabolic effects and anti-tumor properties of cancer cells. These findings illustrate the value of such integrative analysis of transcriptome and metabolic response under various tumor microenvironmental stresses and may lead to important insights on how they influence the phenotypic and metabolic adaptations in human 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 2340.
Abstract 235: Desmocollin 2 (DSC2), identified by CAST method, is associated with intestinal phenotype of gastric cancer
Authors: Anami Katsuhiro , Oue Naohide , Sakamoto Naoya , Hayashi Tetsutaro , Ohara Shinya , Sentani Kazuhiro , Noguchi Tsuyoshi , Yasui Wataru
Published: February 2011

Gastric cancer (GC) is one of the most common human cancers. Identification of novel genes encoding secreted or transmembrane proteins expressed specifically in GC is important for the development of ideal biomarkers for cancer diagnosis and therapeutics. In the present study, we tried to identify genes encoding cell surface and secretory proteins in GC using Escherichia coli ampicillin secretion trap (CAST) method. CAST libraries were generated from two GC cell lines, MKN-1 and MKN-28. Ampicillin-resistant 1,440 colonies from each library were sequenced and these sequences were compared to the public databases using BLAST (NCBI). Many genes encoding secreted and transmembrane proteins including some known genes were identified. Quantitative RT-PCR was performed to confirm the overexpression of the candidate genes in GC, and the expressions of several genes are narrowly restricted in GC. Among these genes, we focused on the desmosomal cadherin desmocollin 2 (DSC2). DSC2 is ubiquitously expressed in several normal tissues where desmosomes exist. In cancers, the reduction of DSC2 in sporadic colorectal cancer is reported as desmocollin switching. It is also known that the homeodomain transcription factors Cdx1 and Cdx2 regulate DSC2 expression in colon cancer cells. Quantitative RT-PCR analyses with 14 kinds of normal tissues and 9 GC cases showed that DSC2 is highly expressed in GCs. In normal tissues, the expression was seen in the heart and pancreas at the lower levels, but not in the stomach. Overexpression of DSC2 was observed in 13 (31%) of 41 GC cases. Immunohistochemical analyses revealed that DSC2 was preferentially expressed in the cases with mucosal or submucosal invasion. DSC2 expression was significantly associated with the well differentiated cases. There were no correlations between DSC2 expression and lymph node metastasis, pathological stage, or overall survival. Additionally, DSC2 was significantly associated in GCs positive for MUC2 and CDX2. Cell invasion assay and MTT assay revealed that knockdown for DSC2 of GC cell lines by siRNA did not inhibit either cell growth or invasion ability. These findings suggest that DSC2 is involved in GC with intestinal differentiation. DSC2 is a good biomarker of intestinal type GC.

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 235.