bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2202: Identification of CDH1 mutations in patients with lobular carcinoma with and without a family history of gastric cancer
Authors: Valente Allyson L. , Ellsworth Rachel E. , Lubert Susan , Holliday Charvonne , Shriver Craig D.
Published: June 2012

Background: The E-cadherin (CDH1) gene is involved in cell adhesion and maintenance of tissue architecture. Loss of expression of CDH1 has been found in both invasive lobular carcinoma (ILCA) of the breast as well as hereditary diffuse gastric cancer (HDGC). While mutations in CDH1 have been found in patients with hereditary diffuse gastric cancer, less is known about the role of CDH1 mutations in patients with breast cancer, thus mutation status was determined in a breast cancer cohort with and without family histories of gastric cancer.

Methods: The Clinical Breast Care Project database was queried to identify all patients with ILCA with or without a family history of gastric or stomach cancer as well as those with non-lobular invasive cancer with a family history of gastric or stomach cancer. Genomic DNA was isolated from peripheral blood samples. DNA variants were detected for each exon using high-resolution melting technology and the underlying change identified by direct sequencing.

Results: Of the 72 patients with ILCA, five had a first degree relative with stomach cancer; 14 patients with non-lobular invasive cancer (12 IDCA, one mixed lobular and ductal, and one tubular) had first degree relatives with stomach or gastric cancer. Three mutations previously associated with gastric cancer were identified: A617T and V832M were identified in a patient with ILCA with no history of gastric cancer and in a patient with IDCA with a family history of gastric cancer, respectively, while one patient with ILCA and no family history harbored the splice site mutation 1137G-T. Three novel variants were also identified: G879S, in a patient with ILCA and no family history, P825Q in a patient with mixed lobular and ductal features with a family history of stomach cancer and L230F in a patient with IDCA and a family history of stomach cancer.

Conclusions: The frequency of CDH1 mutations in this patient population was low (∼7%) and were split between patients with ILCA and no family history of gastric cancer and those with non-lobular cancers and at least one family member with gastric or stomach cancer. Identification of known mutations in patients with non-lobular carcinomas suggests that while these specific CDH1 mutations are associated with increased risk of hereditary diffuse gastric cancer, they do not confer increased risk for lobular cancer. In those patients with ILCA, the low frequency of germline mutations in CDH1 suggests alternate methods, such as epigenetic modification or miRNA silencing, may be driving the suppression of expression of E-cadherin and the development of ILCA.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2202. doi:10.1158/1538-7445.AM2011-2202
Abstract 2204: Loss of heterozygosity in ameloblastic odontogenic tumors
Authors: Gomes Carolina C. , Galvão Calrice F. , Diniz Marina G. , De Marco Luiz , Macedo Paula S. , Vargas Pablo A. , Taylor Adalberto M. , Gomez Ricardo S.
Published: June 2012

The interrelationship of the odontogenic lesions consisting of odontogenic epithelium with odontogenic ectomesenchyme has not been established. To give further insight into the molecular pathogenesis of these odontogenic tumors, the present study investigated allelic loss of a panel of tumor suppressor genes. Six samples of ameloblastic fibroma (AF), 3 cases of ameloblastic fibro-odontoma (AFO) and 3 ameloblastic fibrosarcomas (AFS) were included in the study. We observed loss of heterozygosity (LOH) in 2/6 samples of AF in at least one of the analyzed markers and loss in all the three samples of AFO and AFS in more than one marker. Results are shown in table 1. For the markers AFM238WF2(CHRNB1) and P53, LOH was found all the three types of lesion studied. However, the markers D3S1293, D9S157 and D9S171 showed LOH only in AFS and none of the samples included in the study showed LOH of D11S1369 and D3S1029 markers. LOH was observed for the marker D9S162 in only one sample of AFO. Our data show that AFS have distinct patterns of LOH in tumor suppressor genes compared to their benign counterparts and allelic loss of some markers may be related to tumor behavior, as AFS is a malignant odontogenic tumor and the others are benign entities.

This study was supported by FAPEMIG and CAPES, Brazil.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2204. doi:10.1158/1538-7445.AM2011-2204
Abstract 2205: Inactivation of the Gastrokine 1 gene in gastric adenomas and carcinomas
Authors: Yoon Jung Hwan , Song Jae Hwi , Choi Yoo Jin , Park Won Sang
Published: June 2012

Gastrokine 1 (GKN1) plays an important role in the gastric mucosal defense mechanism and also acts as a functional gastric tumor suppressor. The aim of this study is to determine whether inactivation of the GKN1 gene is involved in the development and/or progression of the gastric cancers. GKN1 expression was examined in gastric adenomas and cancer by immunohistochemistry and Western blot analysis. We also analyzed mutation and epigenetic alteration, DNA copy number change and mRNA transcript of GKN1 in gastric adenomas and carcinomas. The effect of GKN1 on cell proliferation and death was further examined in wild-type GKN1 transfected AGS gastric cancer cells. In immunohistochemistry, reduced or loss of GKN1 expression was detected in 36 (90%) and 170 (89.5%) of 40 adenomas and 190 gastric cancers, respectively. Statistically, there was no significant relationship between altered expression of GKN1 protein and clinicopathologic parameters, including depth of invasion, location, and lymph node metastasis (Chi-Square test, P>0.05). In Western blot analysis, absence or reduced expression was found in 21 (84.0%) of 25 gastric carcinomas. No mutation was detected in gastric tumors, and hypermethylation of GKN1 gene was found in 2 gastric cancers. Interestingly, DNA copy number and mRNA transcript of GKN1 were significantly decreased in gastric cancers. In functional analysis, AGS gastric cancer cells transfected with GKN1 wild-type showed marked inhibition of cell proliferation and induction of cell death. These data suggest that inactivation of the GKN1 gene may play an important role in the development of sporadic gastric cancers, as an early event.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2205. doi:10.1158/1538-7445.AM2011-2205
Abstract 2232: Effectiveness evaluation of an in vitro nucleic acid amplification test for quantification of BCR-ABL fusion transcript variants in human whole blood
Authors: Stanoszek Lauren , Blomquist Thomas , Crawford Erin L. , Austermiller Bradly , Spitzer Casey , Willey Paige FS , Willey James C.
Published: June 2012

Quantitative measurement of BCR-ABL fusion gene transcripts in whole blood by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is a clinically validated method to monitor tyrosine kinase inhibitor drug response in patients diagnosed with chronic myelogenous leukemia (CML). Monitoring of BCR-ABL expression in blood of cML patients under treatment with a tyrosine kinase inhibitor of BCR-ABL is considered standard of care. A three log or greater reduction in BCR-ABL expression from baseline is considered a Maximum Molecular Response (MMR). A subsequent 0.5 log increase in BCR-ABL from MMR is considered a sign of treatment failure sufficient to trigger change in treatment. The challenge is to develop a test with sufficient analytical performance characteristics and quality control to reliably measure BCRABL in MMR samples in which there may be less than 10 molecules in 300 of RNA extracted from whole blood. A method validation protocol was developed using the International Committee on Harmonization (ICH) Q2(R1) international validation guidelines. BCRABL variants b2a2 and b3a2 as well as the housekeeping gene GUSB were measured in KCL22 cell line RNA and Stratagene Universal Human Reference RNA test articles in extreme linear dilution assay, robustness testing conditions, and in multiple laboratories. Quality controls were developed to enable loading as much whole blood RNA into the assay as possible without assay interference. These included a reverse transcription (RT) standardization mixture comprising External RNA Control Consortium (ERCC) reagents to control for RT interference, and controls for genomic DNA contamination, RNA integrity, and PCR interference. Results: This test for b2a2 and b3a2 variants of BCR-ABL meets ICHQ2(R1) guidelines for specificity, linearity, accuracy, LOD and LOQ, imprecision, robustness, and reproducibility. Each analyte comprised by the test was linear (R2 > 0.97) over more than 3 logs10, and demonstrated inter-experimental and inter-laboratory imprecision low enough to enable measurement of a 3-fold difference as significant (P <0.05) from a value as low as five molecules/assay. Experiments are underway to quantify the amount of whole blood RNA that can be loaded into RT reaction without interference.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2232. doi:10.1158/1538-7445.AM2011-2232
Abstract 2244: The M-SKIP Project: an international pooled-analysis on melanocortin-1 receptor (MC1 R) variants and skin carcinogenesis
Authors: Gandini Sara , di Pietro Alessandra , Maisonneuve Patrick , Autier Philippe , Fargnoli Maria Concetta , García-Borrón José C. , Han Jiali , Kanetsky Peter A. , Landi Maria Teresa , Little Julian , Newton-Bishop Julia , Sera Francesco , Raimondi Sara
Published: June 2012

Background: MC1R is one of the major genes involved in the determinism of skin pigmentation. The role of MC1R variants in skin carcinogenesis was previously investigated by a meta-analysis of 18 published studies. Although the results were informative, published data were not enough to deeply investigate the specific contribution of each MC1R variant on skin cancer development.

Aim: The M-SKIP Project aims to perform an international collaborative pooled-analysis on the role of MC1R variants in skin carcinogenesis, using individual patient data. The main endpoint is skin cancer. Secondary endpoints are phenotypic characteristics involved in skin carcinogenesis, e.g. freckling.

Methods and Patients: An Advisory Committee of investigators with great expertise in melanoma and genetic research was established. The authors of appropriate papers were invited to provide their original published and unpublished data.

Preliminary results: Up to February 2010, 27 out of 35 contacted investigators (77%) agreed to join the M-SKIP Project. Thanks to the positive response obtained up to now, we estimate to receive data on around 6,500 melanoma cases, 1,500 non-melanoma skin cancer cases and 13,000 controls. So far, we have already received data on 2,683 melanoma cases, 1,364 non-melanoma skin cancer cases and 4,340 controls from 15 investigators internationally. After quality controls, data are inserted in a central database, and pooled-analysis will be performed in order to evaluate: (1) the association between MC1R variants and skin cancer overall, and by skin cancer type; (2) the association between MC1R variants and phenotypic characteristics under study; (3) MC1R-phenotype interaction in skin cancer risk. In addition, stratified analysis will be performed to understand the independent and dependent role of MC1R variants in skin cancer development by phenotypic characteristics.

Conclusion: The pooled analysis approach applied in such a setting is particularly useful because it will help reaching firm conclusions even in sub-group and interaction analyses, controlling for important confounders.

This data-bank will represent a reference and guide for all investigators working in this research field. Collaboration within investigators would significantly improve the knowledge and accelerate our understanding of the role of MC1R variants in skin carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2244. doi:10.1158/1538-7445.AM2011-2244
Abstract 2254: Nuclear and cytoplasmic epidermal growth factor receptor and p16 expression as prognostic biomarkers in head and neck squamous cell carcinoma
Authors: Husain Hatim , Psyrri Amanda , Slebos Robbert , Ross Douglas T. , Burtness Barbara , Chung Christine H.
Published: June 2012

Introduction: Epidermal growth factor receptor (EGFR) and p16 staining have demonstrated to be strong prognostic factors in patients with head and neck squamous cell carcinoma (HNSCC). Patients with higher expression of EGFR in their tumors have been correlated with decreased survival and shorter time to recurrence. Patients with p16-positive (+) tumors [a surrogate marker of human papillomavirus (HPV) infection] have improved survival compared to p16-negative (−) tumors. We examined the interaction between the expression levels of total EGFR and p16 status, and further examined the association between nuclear EGFR and p16 status based on evidence that nuclear EGFR may have a distinct role in DNA damage repair and may function as a transcription factor.

Methods: A tissue microarray (TMA) containing 123 cores obtained from 101 HNSCC tumors was analyzed for EGFR expression (clone H11; DAKO, Carpinteria, CA) using an automated quantitative analysis (AQUA), and for p16 expression (clone E6H4; MTM Laboratories Inc., Westbrough, MA) using immunohistochemical (IHC) staining. Duplicate cores from a patient were averaged for analyses. The EGFR and p16 data were correlated with clinical data including demographics, primary subsite, disease stage, and survival.

Results: The total and nuclear EGFR data could be reliably obtained from 80 of 101 patients. The p16 data could be obtained from 87 of 101 patients [p16 (+), n=29; p16 (−), n=58]. The p16 staining was strongly associated with the oropharyngeal subsite (Chi2, p=<0.0001) and poorly differentiated histology (Cochran-Armitage Trend Test, p=0.01). The expression of total and nuclear EGFR was higher in p16 (−) tumors compared to p16 (+) tumors (Wilcoxon Rank Test, p=0.038 and p=0.014, respectively). Interestingly, the higher expression of non-nuclear EGFR (combined cytoplasmic and membranous) staining did not associate significantly with p16 (−) tumors. Statistically significant differences in recurrence-free survival or overall survival were not detectable based on the EGFR and p16 expression, likely due to the small sample size and heterogeneity in the patient population.

Conclusions: The total and nuclear EGFR expression levels were higher in p16 (−) tumors compared to p16 (+); this may relate to recurrence and poor survival. Additionally, localization of EGFR may be a significant factor in its association with clinical outcomes. Further studies are indicated to determine the mechanistic link between these two prognostic factors and define the significance of EGFR localization to nucleus upon activation of the pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2254. doi:10.1158/1538-7445.AM2011-2254
Abstract 2285: Stat3 modulates heregulin (HRG)-induced progesterone receptor (PR) transcriptional activation
Authors: Proietti Cecilia J. , Izzo Franco , Flaqué María Celeste Díaz , Vicario Rocío , Tkach Mercedes , Rivas Martín A. , Charreau Eduardo H. , Schillaci Roxana , Elizalde Patricia V.
Published: June 2012

The signal transducer and activator of transcription (Stat) family of proteins was found to be involved in crosstalks with both steroid hormones and type I receptor tyrosine kinases (RTKs) signaling pathways. We have previously demonstrated that HRG, a ligand of RTKs, transactivates PR both in C4HD cells from an experimental model of hormonal carcinogenesis in which MPA induced mammary adenocarcinomas in Balb/c mice, and in the human breast cancer cell line T47D (Mol Cell Biol. 2003 Feb;23(3):1095-111). We have also shown that HRG induces Stat3 tyrosine phosphorylation and transcriptional activation by a mechanism that requires PR signaling (Mol Cell Biol. 2009 Mar;29(5):1249-65).

In the present work we explored whether Stat3 acting as a coactivator, could modulate ligand-independent activation of PR by HRG. Assessment of the expression of the progestin-regulated gene bcl-X showed that HRG treatment of C4HD cells resulted in an increase in bcl-X protein levels. This effect was completely abolished when Stat3 expression was silenced using siRNAs. HRG treatment of cells transfected with a luciferase reporter plasmid under the control of the murine bcl-X promoter which contains two progesterone response elements (PREs), resulted in an increase in luciferase activity. HRG had no effect on PR transcriptional activation when cells were transfected with a mutant vector containing a deletion spanning both PREs in bcl-X promoter. We assessed the specific association of Stat3 and PR to the PRE region of bcl-X promoter in the context of living cells by performing Chromatin Immunoprecipitation (chIP) Assays. We found that HRG treatment of primary cultures of C4HD cells induced PR and Stat3 recruitment to the bcl-X promoter. HRG also induced PR and Stat3 occupancy of the stably integrated MMTV promoter in T47D-Cat0 breast cancer cells. By performing sequential chIP assays we showed that HRG induced simultaneous PR and Stat3 occupancy of the bcl-X promoter region. We explored Stat3 role in the non-classical mechanism of action of PR, where PR is recruited to p21 promoter indirectly through interaction with Sp1 transcription factor. Interestingly, when cells were treated with HRG, we detected Stat3 binding to the Sp1 binding sites of the PR-regulated p21 promoter, together with Sp1 and PR. These results provide the first evidence that Stat3 modulates ligand-independent activation of PR by HRG in breast cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2285. doi:10.1158/1538-7445.AM2011-2285
Abstract 2302: Detection of HMTV env sequences in hormonally dependent tissues
Authors: Pogo Beatriz GT , Moran Heberth , Marin Tania , Lee Anna , Deligdisch Liane , Ancaro Kathleen , Anderton Douglas , Melana Stella , Holland James F.
Published: June 2012

Breast cancers discovered during pregnancy or lactation (gestational) are known to be very aggressive and of poor prognosis. We have found a Mouse Mammary Tumor Virus (MMTV) like virus in human breast cancer, but not in adjacent normal tissue, indicating it exogenous origin. This virus, Human Mammary Tumor Virus (HMTV) recognized by its unique env sequence is present in 62% of gestational breast cancer, compared with 38% of sporadic breast cancers (Wang et al, 2003. Medical Oncology 20:233-236). The aggressive phenotype of gestational breast cancer was suggested to be due to the effect of hormones that stimulate cellular growth. HMTV- LTR contained sequences which are responsive to several hormones (glucocorticoid, estrogen, prolactin, and progesterone) and a transcription factor (TEF-1 family) (Wang et al, 2003. Medical Oncology 20:233-236).

To find if HMTV is present in other tissues that respond to hormones, we have investigated the presence of HMTV sequences in two of these tissues. Endometrium is a classical example of such tissue. Endometrial cancers and normal endometrium were analyzed for HMTV env sequences. The results indicated that HMTV env sequences were present in 3.4% of the normal tissues and 37% of the tumors analyzed. Recently, it was reported that MMTV-like sequence was found in 10% of endometrial cancers (Johal et al. 2010, J. Med. Virology; 82: 1044-50).

During lactation, the mammary gland experiences major metabolic changes due to hormonal stimulation. We have examined 88 normal milks and 63 milks from women, who have undergone biopsies. Whereas in the cells of normal milks 8% showed env sequences cells in the milk of the biopsied group showed that 23.8% contain HMTV sequences.

These results confirmed electron microscope observations that a betaretrovirus was present in milk from healthy women and those with breast cancer (Moore et al. 1971, Nature 229: 611-615).

Taken together, the results presented here suggest that tissues and tumors stimulated by hormones showed high prevalence of HMTV sequences.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2302. doi:10.1158/1538-7445.AM2011-2302
Abstract 2305: Role of ST3Gal1 sialyltransferase in breast cancer cells
Authors: Fan Tan-chi , Lin Wen-der , Chang Chih-hao , Chang Lan-yi , Chen Zhin-mei , Khoo Kay-hooi , Yu Alice
Published: June 2012

Alteration of glycan structures on glycoproteins or glycolipids is a common phenomenon observed in malignancy. The premature sialylation of core carbohydrate structure leading to shorter truncated glycans has been well documented. Mucin O-glycans from normal breast are mostly composed of elongated core 2 structures. It has been shown that MUC1 O-glycans from breast cancer cell lines are often truncated and mainly consist of core 1-based structures from mass spectrometry sequencing. It may be due to down-regulated C2GnT (core 2 enzymes) or increased sialyltransferase activities which add sialic acid in an α2,3 linkage to Galβ1-3GalNAc, producing sialyl3 T antigen, which is no longer recognized by C2GnT as a substrate donor. Six different α2,3-sialyltransferases catalyze the transfer of sialic acids to Gal. Among which, ST3Gal1 and ST3Gal2 are capable of producing sialyl3 T antigens; however, only the expression of ST3Gal1 mRNA is elevated in primary breast carcinoma cells compared to normal or benign breast tissues. In ductal carcinomas, the expression of ST3Gal1 appeared to correlate with histology grade. However, not much is known about the role of ST3Gal1 in breast cancers. We investigated the role of ST3Gal1-mediated sialylation in the function of proteins and further characterize the molecular mechanism underlying ST3Gal1-mediated cancer metastasis.

To confirm the relationship of ST3Gal1 expression in breast cancers, we performed qPCR analysis of sialyltransferase gene expression in 93 breast cancer patients. ST3Gal1 expression was statistically higher in ER-, PR-, and HER2-negative category (p<0.037). Using MALDI-MS analysis, the glycome profiles showed that ST3Gal1 silencing in MDA-MB-231 breast cancer cells resulted in reduced α2,3-linked sialic acid-containing O-glycosylation including sialyl3T antigen, without significant differences in N-glycosylation. We observed that ST3Gal1 silenced MDA-MB-231 cells have the tendency to aggregate compared to control cells. To test the hypothesis that decrease of sialylation in beast cancer cells will affect the metastatic behavior, cell migration and invasion were assayed. Indeed, Transwell migration of ST3Gal1 silenced MDA-MB-231 significantly decreased to ∼30% of control cells. More strikingly, the Transwell invasion of ST3Gal1 silenced cells were down-regulated to less than 10% of control cell. Furthermore, the enzyme activities of MMP9 and m-calpain which are known to regulate the invasion ability of cancer cells were both down-regulated upon silencing of ST3Gal1. In conclusion, these data strongly suggest that cell surface α2,3-sialylation contributes to breast cancer cells invasiveness.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2305. doi:10.1158/1538-7445.AM2011-2305
Abstract 2316: Therapeutic targeting of novel human stem cell and cancer associated glycans
Authors: Saunders Mary R. , An Hyun Joo , Olivos David J. , Maeda Yoshiko , Lim Ryan G. , Lebrilla Carlito B. , Lam Kit S.
Published: June 2012

Host and tumor surface glycans are important in regulating pivotal pathophysiological events during the development of cancer and tumor progression. Changes in glycosylation allow tumorigenic cells to seize control over developmental processes which allow cells to proliferate, invade, promote angiogenesis, and metastasize. Malignant transformation is accompanied by the expression of oncofetal antigens which are expressed on embryonic cells, tumor cells, and a few adult cells. Glycans common to human embryonic stem cells and cancer cells can be targeted using combinatorial chemistry. Several peptidomimetic ligands have been discovered using One Bead One Compound (OBOC) and One Bead Two Compound (OB2C) methods. These cell adhering ligands possess the potential to target tumorigenic cells and mediate intracellular signaling events by binding to high mannose glycans expressed on the surface of Jurkat, TK6, Her2 expressing MCF7, chemoresistant MCF7, HeLa, and SiHa cell lines. Three lead linear peptides with alternating natural and unnatural amino acids have been investigated for their in vivo targeting potential in the lymphoid leukemia, breast, and cervical cancer mouse xenograph models. In vitro and in vivo stem and cancer cell experiments, which demonstrate genetically altered glycan function, are currently being investigated for their anticancer potential. These novel agents may be used alone or in combination with chemoradiation and anti-angiogenesis strategies for treating cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2316. doi:10.1158/1538-7445.AM2011-2316
Abstract 2328: 15N-backbone and side-chain dynamics of the mutant calcium-binding protein S100B D63N
Authors: Liriano Melissa , Varney Kristen M. , Ishima Rieko , Weber David J.
Published: June 2012

Similar to calmodulin and other EF-hand-containing proteins, S100 proteins have no inherent enzymatic activity and perform their biological functions via calcium-dependent protein-protein interactions. Elevated protein levels of S100B have been linked to various types of cancers, including malignant melanomas, glioblastomas, and anaplastic astrocytomas. In the case of malignant melanomas, S100B contributes to cell proliferation by binding to p53 and inhibiting its tumor suppressing properties. However, one property of S100 proteins, including S100B, is that it only binds to calcium tightly when its target protein is bound, a feature that drug design studies for restoring p53 in melanoma must mimic. Therefore, the purpose of this study is to determine the mechanism for how target protein binding to S100B causes an increase in calcium-binding affinity. Previous studies suggest that the increase in calcium affinity in the presence of target is not the result of a structural modification. As an alternative explanation, the mechanism behind this event may involve a reduction in the ligand dynamics coordinating calcium in the canonical EF-hand when target peptide is present. To test this hypothesis with S100B, an asparagine mutant was made in order to have an 15N-probe in the tight-binding EF-hand of S100B for NMR side-chain dynamics with and without target peptide. 15N backbone dynamic experiments (T1, T2, and hetNOE) were collected for the S100B D63N mutant (+/- TRTK-12) at two magnetic fields to determine the effects of peptide binding at multiple time scales. Relaxation dispersion experiments were also collected for backbone and side-chain amides of S100B in the presence and absence of TRTK-12. For the backbone dynamic experiments, motion was detected in the hinge region and C-terminus of helix-4 at multiple timescales in the absence of TRTK-12 (ns-sec timescale). A similar profile was observed for those backbone residues undergoing motion in the chemical exchange time scale seen with the relaxation dispersion data. However, when a molecular target was introduced, the majority of these backbone amides that make up the hydrophobic pocket displayed a general decrease in dynamics. Side chain dynamics in the EF-hand was appreciated in the absence of TRTK-12 peptide, however when the target was present, no chemical exchange was seen with the 15N-D63N probe in the EF-hand. These findings imply that In the presence of target peptide, S100 proteins undergo a decrease in protein dynamics that translates from the site of target binding to the EF-hand.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2328. doi:10.1158/1538-7445.AM2011-2328
Abstract 2330: Role of RET finger protein in the integrin beta1 expression in cancer cells and its significance in cancer patients
Authors: Kato Takuya , Enomoto Atsushi , Asai Naoya , Murakumo Yoshiki , Takahashi Masahide
Published: June 2012

Integrins play central roles in the biology by regulating cell adhesion to the extracellular matrix and cell migration, growth, differentiation and apoptosis. Although contribution of integrins on cancer development has been well studied, the mechanism of the regulation of integrins expression remains elusive. Here we report that RET finger protein (RFP) regulates integrin beta1 expression in cancer cells. RNAi-mediated depletion of RFP resulted in decrease of integrin beta1 expression and impaired cell migration/invasion. RNA immunoprecipitation assay revealed that RFP interacts with integrin beta1 mRNA, and RFP knockdown caused destabilization of the mRNA. Furthermore, we examined the expression of RFP in human colon cancer and endometrial cancer, and assessed its clinical significance. High levels of RFP expression correlated with poor clinical outcome in colon cancer and endometrial cancer patients. These findings suggest that RFP regulates the cancer cell migration/invasion through the upregulation of integrin beta1 expression by stabilization of its mRNA, and expression of RFP can be a prognostic marker for an unfavorable clinical outcome in patients with colon cancer and endometrial cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2330. doi:10.1158/1538-7445.AM2011-2330
Abstract 2336: SFRP4 expression as a marker of invasion in gastric cancer
Authors: Busuttil Rita A. , George Joshy , Scott Kylie , Tan Patrick , Lade Stephen , Bowtell David D. , Boussioutas Alex
Published: June 2012

Gastric cancer (GC) is the fourth most common cancer in the world with approximately 1 million patients being diagnosed annually and is the second highest cause of cancer related deaths worldwide resulting in 700,000 deaths per year. The advanced stage at diagnosis and limited curative treatments for this stage of disease explains the mortality rate for GC. Early and late GC can usually be differentiated by their T-stage, which is a measure of invasion of the tumour through the gastric wall. T1 tumours exhibit minimal invasion whilst T4 tumours have invaded into adjacent structures.

Through our genomic research we have identified SFRP4 as a gene that is consistently over-expressed in gastric cancer when compared to normal gastric tissue, particularly advanced stage cancer. Its gene expression levels are directly proportional to the degree of invasion suggesting a potential role in this process. Analysis of the clinical data in conjunction with the genomics data shows that elevated levels of SFRP4 have also been correlated with poor prognosis in patients with GC independent of their T-stage.

We have used shRNA knockdown of SFRP4 in a gastric cell line to validate these findings in vitro. The invasive phenotype is able to be restored in these cells with the addition of recombinant SFRP4 protein. Evidence from this study suggests that SFRP4 plays a role in invasion which is a fundamental property of all cancers. In vivo studies are currently underway to determine whether the functional effects of SFRP4 observed in vitro in gastric cancer lines can be recapitulated in vivo.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2336. doi:10.1158/1538-7445.AM2011-2336
Abstract 2345: Identification of epithelial stromal interaction 1 as a novel target of KLF8
Authors: Li Tianshu , Lu Heng , Wang Xianhui , Yu Lin , Zhao Jihe
Published: June 2012

Epithelial stromal interaction 1 (EPSTI1) is a recently identified novel gene with two splicing variants that is highly upregulated in breast cancer cells when co-cultured with the stromal fibroblasts and in invasive breast carcinomas as compared with normal breast biopsies. The regulation and function of EPSTI1 are largely unknown. Interestingly, our microarray studies showed that EPSTI1 is one of the most upregulated genes in MCF-10A cells expressing ectopic Kruppel-like factor 8 (KLF8), a transcriptional factor is highly over-expressed in breast cancer and critical for cell cycle progression, oncogenic transformation, epithelial to mesenchymal transition (EMT) and metastatic progression of breast cancer. Real time qRT-PCR and western blotting demonstrated that EPSTI1 mRNA and protein levels are upregulated in a panel of invasive breast cancer cell lines that are known for aberrant overexpression of KLF8 as compared to noninvasive cell lines. Promoter reporter analysis indicated that KLF8 may activate EPSTI1 gene promoter. These results identified EPSTI1 as a novel target of KLF8 and suggest that EPSTI1 may play an important role for KLF8 signaling in the regulation of breast cancer invasion and metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2345. doi:10.1158/1538-7445.AM2011-2345
Abstract 2357: The TROY/TNFRSF19 receptor is overexpressed in human glioblastoma multiforme and regulates glioma cell survival via the NFkB pathway activation
Authors: Tuncali Serdar M.
Published: June 2012

Glial tumors progress to malignant grades by heightened proliferation and aggressive invasion of the surrounding normal brain. Understanding the genetic and biochemical processes that foster these behaviors is likely to reveal specific and effective targets for therapeutic intervention. We have identified TROY / TNFRSF19 as a novel transmembrane receptor that is overexpressed in glioblastoma multiforme. TROY is an orphan member of the tumor necrosis factor receptor (TNFR) superfamily that lacks a cytoplasmic death domain. Our previous data demonstrates that the level of TROY mRNA expression directly correlates with increasing glial tumor grade and inversely with overall patient survival. In addition, TROY overexpression in glioma cells activates Rac1 signaling in a Pyk2-dependent manner to drive invasion and migration. Here, we show that TROY over-expression enhances glioma cell survival against chemotherapy-induced cell death. TROY expressing glioma cells also displayed an increased resistance to temozolomide (TMZ) treatment. Conversely, siRNA mediated-depletion of TROY expression enhanced TMZ-induced cell death. To investigate the mechanism of TROY-induced cell survival, we surveyed a panel of known cancer signaling pathways by immunoblot analysis of cell lysates from untransfected glioma cells and lysates from glioma cells overexpressing TROY. Of the signaling pathways surveyed, we observed an increased activation of the Akt and NFκB pathways in TROY overexpressing cells relative to the untransfected cells. Inhibition of the NFκB pathway by expression of the super-repressor IκBα mutant abrogated TROY-induced cell survival to TMZ treatment. Similarly, inhibition of the Akt pathway by treatment of cells with the pharmacological inhibitor, LY294002, increased TMZ-induced cell death in TROY expressing cells. Together, these data indicate that TROY-stimulated glioma cell migration and invasion increases resistance to chemotherapy-induced cell death in glioma. Understanding the function of TROY in glioma biology may lead to the development of effective therapies against invasive gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2357. doi:10.1158/1538-7445.AM2011-2357
Abstract 2358: Molecular chaperones as a common set of proteins that regulate the invasion phenotype of head and neck cancer
Authors: Chiu Ching-Chi , Cheng Ann-Joy
Published: June 2012

Purpose: The goal of this study was to establish a common set of molecules that regulate cell invasion in head and neck cancer (HNC). Experimental design: Five invasive sublines derived from HNC cell lines were established using the Matrigel selection method. Proteomic technology, MetaCore™ algorithm, and RT-PCR methods were used to search for molecules that contribute to the invasion phenotype. Cellular functional analyses and clinical association studies were applied to examine the significance of the molecules. Results: Fifty-two proteins were identified in more than two of the four independent proteomic experiments, including 10 (19%) molecular chaperones. Seven chaperones were confirmed to be differentially expressed in five sublines: Hsp90α, Hsp90β, Hsp90-B1/Gp96, Hsp70-A5/Grp78, HYOU1 up-regulation, while Hsp60 and GANAB down-regulation. Four molecules with the highest differential expressions (> 4-fold) were further investigated. In all cell lines, knockdown of Hsp60 or GANAB and silencing of Gp96 or Grp78 considerably enhanced or reduced cell migration and invasion, respectively. Clinical association studies consistently revealed that low levels of Hsp60 or GANAB and high levels of Gp96 or Grp78 are significantly associated with advanced cancer (P<0.001 to P = 0.047, respectively, for the four molecules) and poor survival (P<0.001 to P=0.025, respectively, for the four molecules). Conclusion: In conclusion, our study defined molecular chaperones as a common set of proteins that regulate the invasion phenotype of head and neck cancer. Loss of the tumor suppression function of Hsp60 or GANAB and acquisition of the oncogenic function of Gp96 or Grp78 contribute to aggressive cancers. These molecules may serve as prognostic markers and targets for cancer drug development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2358. doi:10.1158/1538-7445.AM2011-2358
Abstract 2360: Dimethyl sulfoxide induces upregulation of tumor suppressor HLJ1 through activator protein-1 activation in human lung adenocarcinoma CL1-5 cells
Authors: Wang Chi-Chung , Lin Ai-Me , Lai Yi-Hua , Lin Sheng-Yi , Yang Pan-Chyr , Chen Jeremy J. W.
Published: June 2012

Dimethyl sulfoxide (DMSO) is an amphipathic molecule and has a diversity of antitumor activities. Previous studies demonstrated that DMSO could modulate AP-1 activity and lead to cell cycle arrest at G1 phase. HLJ1 is a newly identified tumor and invasion suppressor that inhibits tumorigenesis and cancer metastasis. Its transcriptional activity is also regulated by transcription factor AP-1. However, the effect of DMSO on HLJ1 is still unknown. In this study, low-HLJ1 expressing highly invasive CL1-5 lung adenocarcinoma cells were subjected to DMSO treatment. We found that DMSO can significantly inhibit cancer cell invasion and migration capabilities through up-regulation of HLJ1 in a concentration-dependent manner. In addition, knockdown of HLJ1 expression by siRNA was able to block the effects of DMSO-induction. The HLJ1 promoter and enhancer reporter assay revealed that DMSO transcriptionally upregulates HLJ1 expression through an AP-1 site within the HLJ1 enhancer. Furthermore, AP-1 subfamily members, JunD and JunB, were significantly upregulated by DMSO in a concentration-dependent manner. In conclusion, our results suggest that DMSO could be an important stimulator of the tumor suppressor protein, HLJ1, through AP-1 activation in highly invasive lung adenocarcinoma CL1-5 cells. Our findings provide some important information for further investigations to develop new drugs such as DMSO-derived analogues which may inhibit cancer invasion and progression by targeting HLJ1.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2360. doi:10.1158/1538-7445.AM2011-2360
Abstract 2379: Cyclin D3 is important for ErbB2-induced mammary cancer initiation and progression in the absence of Cyclin D1
Authors: Zhang Qian , Sakamoto Kazuhito , Triplett Aleata A. , Lin Wan-chi , Creamer Bradley A. , Wagner Kay-Uwe
Published: June 2012

The cell cycle regulatory protein Cyclin D1 has been reported to be essential for both normal mammary gland development during pregnancy and the malignant transformation of the mammary epithelium in response to receptor tyrosine kinase (RTK) signaling. We have recently reported that the prolactin receptor-associated kinase Jak2 controls the transcriptional activation and, more importantly, the nuclear accumulation of Cyclin D1 in normal mammary epithelial cells via expression and activation of Akt1. We also demonstrated that the functional ablation of Jak2 and Stat5 in ErbB2-expressing mammary epithelial cells protects against the onset of mammary tumorigenesis. Jak2, however, is not required for the survival and proliferation of neoplastic cells. The constitutive activation of ErbB2 signaling, which is an initial event in the formation of mammary cancer in this model, was able to override the functional role of Jak2 in regulating the expression of Akt1 and Cyclin D1. To further define the role of the downstream effector Cyclin D1 during cancer initiation and progression in response to Jak2 and RTK signaling, we generated an ErbB2-associated mammary cancer model that allows a functional ablation of Cyclin D1 either prior to or following neoplastic transformation. Surprisingly, and in sharp contrast to previous reports, nearly all Cyclin D1-deficient females expressing wildtype ErbB2 developed mammary tumors. While Cyclin D1 deficiency extended the tumor-free survival, this particular cell cycle regulatory protein was clearly not required for tumor maintenance and progression. Growth and proliferation of the vast majority of Cyclin D1-deficient primary mammary cancers was accompanied by an upregulation of Cyclin D3. Since Cyclin D2 expression remained very low, knocking down the last remaining D-type cyclin (i.e. Cyclin D3) in tumor-derived primary cancer cells using three different shRNA constructs was sufficient to attenuate the proliferation rates. Interestingly, the ablation of the pool of all three D-type cyclins did not result in an upregulation of Cyclin E. Collectively, the results of this study show that Cyclin D1 is dispensable for neoplastic transformation of mammary epithelial cells and it might therefore not serve as a sole therapeutic target to treat ErbB2-associated breast cancers as previously suggested. In addition, these observations indicate that Cyclin D1 is most likely not the only downstream effector of Jak2 signaling that mediates the protective effects of a functional ablation of Jak2 in ErbB2-induced mammary carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2379. doi:10.1158/1538-7445.AM2011-2379
Abstract 2386: Overexpression of 14-3-3ζ in mouse mammary gland promotes mammary tumor development and progression by enhancing multiple tumor-associated properties
Authors: Li Shau-Hsuan , Wyszomierski Shannon L. , Li Ping , Wang Qingfei , Rehman Sumaiyah K. , Xiong Yan , Yang Jun , Guo Hua , Zhang Siyuan , Fang Dexing , Medina Daniel , Muller William J. , Yu Dihua
Published: June 2012

The 14-3-3 proteins are a family of evolutionarily conserved proteins ubiquitously expressed in eukaryotic organisms. 14-3-3 proteins interact with and regulate many different target proteins including oncoproteins and tumor suppressors. Previously, we found that 14-3-3ζ was overexpressed in >40% of human breast cancers and associated with poor clinical outcome of patients. We also showed that 14-3-3ζ contributed to the transformation of MCF10A non-transformed human mammary epithelial cells and multiple breast cancer cells in vitro. Here, to assess the oncogenic capacity of 14-3-3ζ in vivo, we generated transgenic mice with HA-tagged 114-3-3ζ overexpressed in the mammary epithelium, driven by either the whey acidic protein promoter (WAP-HA-14-3-3ζ) or the MMTV- LTR promoter (MMTV-HA-14-3-3ζ).

Multiparous WAP-HA-14-3-3ζ transgenic mice developed hyperplastic lesions and showed an increased susceptibility to the induction of mammary tumors by the carcinogen, DMBA. Remarkably, MMTV-HA-14-3-3ζ mice developed mammary tumors with a median tumor latency of 662 days without DMBA treatment. WAP-HA-14-3-3ζ and MMTV-HA-14-3-3ζ mice crossed with MMTV-neu transgenic mice (WAP-ζ.neu and and MMTV-ζ.neu) exhibited accelerated neu-mediated mammary tumorigenesis and metastasis. Compared to the mammary tumors of MMTV-neu mice, 14-3-3ζ overexpression in bitransgenic mice tumors further increased cell proliferation, reduced apoptosis and enhanced angiogenesis. These data indicate that 14-3-3ζ overexpression in the mouse mammary gland promotes the development and progression of mammary tumors and that 14-3-3ζ, in cooperation with other oncogenic events, plays a causal role in mammary tumorigenesis and metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2386. doi:10.1158/1538-7445.AM2011-2386