bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1692: Expression of argininosuccinate synthetase protein and correlation with response to the novel therapeutic pegylated arginine deiminase (ADI-PEG20) in osteosarcoma
Authors: Sowers Rebecca , Bomalaski John , Gorlick Richard
Published: August 2012

The amino acid arginine is involved in protein synthesis and tumor metabolism and is essential for the growth of human cancer cells. Pegylated arginine deiminase (ADI-PEG20) is a novel therapy that lowers extracellular arginine levels and has shown evidence of clinical response and low toxicity in patients with tumors lacking argininosuccinate synthetase (ASS1) protein expression, including melanoma and hepatocellular carcinoma. Recently it has been shown that ASS1 expression was deficient in 63% (39/62) of Japanese osteosarcoma patient samples, and reduced ASS1 was associated with pulmonary metastases. In addition, they showed that ASS1 deficient, but not ASS1 expressing, osteosarcoma cells lines did not grow in arginine-deficient media, and recommended evaluation of arginine depletion with ADI-PEG20 in osteosarcoma. Because ADI-PEG20 efficacy appears to correlate with ASS1 protein expression, a cohort of osteosarcoma tissue sections (n=171) was stained immunohistochemically to determine the presence or absence of ASS1 protein. Sixty-four percent (n=109) indicated negligible or no staining for ASS1 protein suggesting further that osteosarcoma might be a candidate for ADI-PEG20 sensitivity. Western blots were performed on patient-derived osteosarcoma cell lines (n=18) to determine levels of ASS1 protein expression prior to treatment with ADI-PEG20. Of the 18 cell lines studied, four cell lines had noticeably diminished ASS1 protein expression. The remaining 13 cell lines exhibited varying amounts of ASS1 protein expression with five of these cell lines appearing strongly positive indicating possible overexpression of ASS1 protein. Cytotoxicity assays were performed on a subset of cell lines representing both elevated and decreased ASS1 protein expression to determine the in vitro effectiveness of ADI-PEG20 in patient-derived osteosarcoma cell lines and its correlation with ASS1 protein expression. Some of the ASS1 deficient cell lines showed decreased cell viability in response to arginine depletion with ADI-PEG 20. These findings suggest that arginine depletion with ADI-PEG 20 may be a potential targeted treatment for osteosarcoma patients deficient in ASS1 expression. In addition, further preclinical studies are ongoing, including xenograft models, to understand better the mechanism of this agent in osteosarcoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1692. doi:1538-7445.AM2012-1692
Abstract 1694: Imaging cell cycle progression in gastric cancer lymph node metastasis indicates large fraction of proliferating cancer cells
Authors: Suetsugu Atsushi , Hassanein Mohamed , Moriwaki Hisataka , Yano Shuya , Fujiwara Toshiyoshi , Bouvet Michael , Saji Shigetoyo , Hoffman Robert M.
Published: August 2012

We report here a mouse model of lymph node metastasis of stomach cancer and imaging cell cycle progression in the metastasis. FUCCI (fluorescent ubiquitination-based cell cycle indicator)-expressing human MKN45 gastric cancer cells were used to establish the lymph node metastasis nude mouse model. In this model, cell-cycle dynamics of the metastatic cells could be imaged in vivo. FUCCI cells are labeled orange in the G1 phase nuclei (monomeric Kusabira-Orange2: mKO2) and labeled green in the S/G2/M phases (monomeric Azami-Green1: mAG1). FUCCI MKN45 were injected into the mouse stomach mucosa from the serosal side. Lymph node metastasis to the para-aortic or hepatic portal region, left armpit, and brachial lymph nodes were observed. Green-yellow (G2/S) cells were predominant compared to red (G1) fluorescent FUCCI cells as observed in the lymph-node metastases using an FV1000 confocal microscope, indicating that the lymph-node metastatic cells were actively cycling. The origin of lymph node or vessel metastases was confirmed by H.E. staining. Mouse FUCCI gastric metastasis models will be used to visualize cell cycle dynamics in metastasis in lymphoid as well as hematogenous metastasis in order to optimize drug treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1694. doi:1538-7445.AM2012-1694
Abstract 1697: Gadd45A expression is high in luminal A and her2 human breast cancers, but frequently absent in triple negative breast cancers
Authors: Tront Jennifer S. , Willis Alliric , Huang Yajue , Smith Geoffery , Powers Benjamin , Hoffman Barbara , Liebermann Dan
Published: August 2012

Gadd45a is a stress sensor, playing an important role in tumorigenesis. Generation and side by side analysis of breast cancer prone MMTV-myc and MMTV-ras mice highlight a unique role for Gadd45a as either a suppressor or promoter of breast cancer development, employing distinct signaling pathways in response to distinct oncogenic stress stimuli. Thus, it appears that gadd45a can function to either promote or suppress breast tumor development in mice via engagement of different signaling pathways depending on the molecular nature of the activated oncogene. Extending the work to human breast cancer has provided initial, novel data, showing that Gadd45a, which is not expressed in normal breast tissue, is expressed at high levels in less aggressive breast cancers and is low or absent in more aggressive subtypes. Notably, Gadd45a is expressed at very high levels in luminal A and her2 positive breast cancers, but frequently is absent in triple negative breast cancers. Experiments are currently underway to elucidate the functional role of Gadd45a in human breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1697. doi:1538-7445.AM2012-1697
Abstract 170: Thymoquinone-mediated suppression of NF- αB activity causes inhibition of cell viability and induces apoptosis in activated B-cell sub-type of diffuse large B-cell lymphoma
Authors: Hussain Azhar R. , Ahmed Saeeda O. , Ahmed Maqbool , Khan Omar S. , Uddin Shahab , Al-Kuraya Khawla S.
Published: August 2012

Diffuse Large B-cell Lymphoma (DLBCL) is the most common lymphoid malignancy and constitutes approximately 40% of all cases of non-Hodgkin's lymphoma (NHL). Despite improvement in treatment protocols, a large number of DLBCL cases remain refractory to treatment. DLBCL can be further classified as three distinct subtypes: germinal center B cell-like (GCB)-, activated B cell-like (ABC)-, and primary mediastinal B-cell lymphoma (PMBL). ABC DLBCL has a worse survival after upfront chemotherapy and is characterized by constitutive activation of the anti-apoptotic nuclear factor-kappa B (NF-κB) pathway. Yet the implication of NF-κB inhibition in ABC DLBCL need to be further elucidated. We have previously shown that Thymoquinone (TQ), a natural compound isolated from Nigella sativa, induces growth inhibition and induces apoptosis in several NHL cell lines. However its role in inhibition of growth and apoptosis has not been fully elucidated in ABC sub-type of DLBCL cell lines. We found that TQ treatment inhibits cell viability and induces apoptosis in a dose dependent manner in various ABC cell lines. Further more, we found that TQ treatment causes inactivation of IκBα and consequently inhibits NF-κB activity by decreasing the expression of p65 subunit of NF-κB in the nuclear compartment of ABC cell lines. In-activation of NF-κB survival pathway leads to induction of apoptosis via the mitochondrial apoptotic pathway initiated by conformational changes in the Bax protein leading to changes in the mitochondrial membrane potential and release of cytochrome c in the cytosole. Once cytochrome c is released, it leads to activation and cleavage of caspases-9 and -3 and cleavage of PARP. This leads to caspase dependent apoptosis in ABC cell lines. In addition, we also found that TQ treatment causes up-regulation of death receptor 5 (DR5), an important and essential receptor for tumor necrosis factor related apoptosis inducing ligand (TRAIL). However, up-regulation of DR5 does not play a role in TQ-induced apoptosis in ABC cell lines. Interestingly, combination of sub-toxic doses of TQ and TRAIL induces efficient significant apoptosis in ABC cell lines. These data show that TQ can be used alone or in combination with TRAIL to induce apoptosis in these aggressive sub-types of DLBCL and may play an important role in the management of DLBCL in the future.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 170. doi:1538-7445.AM2012-170
Abstract 1701: Clinical significance of serum and tissue Dickkopf-1 in patients with gastric cancer
Authors: Park Kyoung Un , Park Do Joong , Kim Hyung Ho , Kim Woo Ho , Lee Hye Seung
Published: August 2012

Background: Dickkopf-1 (DKK-1) is a secreted protein known as a negative regulator of the Wnt signaling pathway and involved in embryonic development and tumorigenesis. Clinical significance of serum DKK-1 and tissue DKK-1 expression in gastric cancer has not been clarified. Methods: Serum concentrations of DKK-1 were assessed by enzyme-linked immunosorbent assay in 153 gastric cancer patients and 33 healthy controls. We also obtained serum CEA (ng/mL) and CA19-9 (U/mL) values by chemiluminescence immunoassay. For tissue DKK-1, we performed immunohistochemical staining for DKK-1 in 144 cancer specimens of 153 patients and in additional 265 consecutive gastric cancer specimens. Results: Serum DKK-1 concentrations were significantly higher in gastric cancer patients (56.27 pg/mL) than in healthy controls (30.50 pg/mL, p=0.018). With 36.3150 pg/mL of cut-off value, sensitivity and specificity for gastric cancer diagnosis were 80.4% and 78.8%, respectively. Gastric cancer patients with a serum DKK-1 of 60.0 pg/mL or higher had a significantly diminished survival compared with patients whose serum DKK-1 were lower (p=0.033). By immunohistochemistry, DKK-1 overexpression in cancer specimens was significantly correlated with increased serum DKK-1 concentrations (correlation coefficient, 0.395; p<0.001). The cancer patients with DKK-1 overexpression had a significantly worse prognosis than the patients without DKK-1 expression in this cohort (p=0.004) and additional consecutive cases (p=0.030). By multivariate Cox regression model, DKK-1 expression status predicted patient prognosis independently of pTNM stage and histologic differentiation in consecutive cases (p=0.018). Conclusion: Increased serum DKK-1 concentrations and overexpression of DKK-1 in gastric cancer tissue may contribute to diagnosing gastric cancer and predicting cancer patient outcome.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1701. doi:1538-7445.AM2012-1701
Abstract 1702: A novel diagnostic approach using genome-wide single nucleotide polymorphism arrays for ovarian metastases in endometrial cancers
Authors: Ikeda Yuji , Oda Katsutoshi , Nakagawa Shunsuke , Hosokawa Satsuki Murayama , Shoji Keiko , Yamamoto Shogo , Ishikawa Shumpei , Uehara Yuriko , Koso Takahiro , Miyasaka Aki , Kashiyama Tomoko , Takazawa Yutaka , Maeda Daichi , Wada Osamu Hiraike , Kawana Kei , Fukayama Masashi , Aburatani Hiroyuki , Yano Tetsu , Kozuma Shiro , Taketani Yuji
Published: August 2012

[Introduction] Synchronous cancers involving ovaries as well as uterine corpus are well-known events in gynecologic malignancies. These tumors may be independently derived, non-metastatic tumors (Dual Primary tumors; DP) or a tumor from one organ with metastasis to the other (Single Primary tumor with Metastasis; SPM). In the majority of these synchronous endometrial and ovarian cancers, both tumors are histologically endometrioid adenocarcinomas, which may cause a diagnostic difficulty to distinguish DP and SPM. By genome-wide genotyping, single nucleotide polymorphism (SNP) arrays, we can obtain chromosomal copy number alterations (CNA) throughout the genome in a single assay. We hypothesized that the copy number profiles by SNP arrays might be highly informative for genetic diagnosis in the synchronous endometrioid adenocarcinomas. [Material and Methods] We genetically diagnosed ten tumors from five patients with synchronous endometrial and ovarian carcinomas, using 250K SNP typing arrays and mutational analysis of PIK3CA, PTEN, K-Ras and CTNNB1 (beta-catenin). We evaluated whether conventional pathological diagnosis is compatible with the genetic diagnosis. [Results] Three of the five patients show identical copy number alterations (CNA), including type, loci and degree of each alteration, between the endometrial and the ovarian carcinomas. The other two show CNA only in either endometrial or ovarian carcinoma. All the five tumors possess one or more genetic mutations in the genes examined. One patient showed mutations both in PIK3CA and PTEN at discordant sites between endometrial and ovarian carcinomas, whereas the other four showed concordant mutations. Together, four of the five were genetically diagnosed as SPM and the remaining one was as DP. The pathological diagnosis was not in agreement with the molecular diagnosis in four of the five cases. [Conclusion] Our data suggest that most of the copy number alterations might occur before metastases and that genome-wide genotyping may represent a useful approach to distinguish between SPM and DP in synchronous endometrial and ovarian carcinomas. As chromosomal instability is commonly observed in various types of tumors, SNP array genotyping might be applicable to synchronous tumors in other organs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1702. doi:1538-7445.AM2012-1702
Abstract 1709: The effect of a XIAP inhibitor, embelin, on apoptosis of thyroid cancer cell lines
Authors: Han Ji Min , Kim Won Bae , Yim Ji Hye , Kim Won Gu , Kim Tae Young , Choi Hyun-Jeung , Kim Eui Young , Shong Young Kee
Published: August 2012

X-linked inhibitor of apoptosis protein (XIAP) is a member of the inhibitor of apoptosis (IAP) protein family that selectively blocks caspases-3, -7, -9 and inhibits cell death. Embelin, a XIAP inhibitor, is known to exhibit anti-inflammatory and apoptotic activities. Recent reports suggest that embelin interferes the signal transducer and activator of transcription 3 (STAT3) pathway and nuclear factor-kB (NF-kB) signal pathway. In this study, we investigated the anti-cancer efficacy of embelin by measuring its effects on apoptosis of thyroid cancer cell lines. We also explored the mechanism underlying these effects. We found that embelin induced the apoptosis of human thyroid cancer cell lines, such as FTC-133, CAL-62, KTC-1, and 8505C. The effect was stronger in BRAF V600E-mutant thyroid cancer cell lines (KTC-1 and 8505C) than wild-type cell lines (FTC-133 and CAL-62). The effect of embelin on cell apoptosis was associated with increased phosphorylation of p38. SB203580 inhibited the embelin mediated induction of cleaved caspase-3 activity in V600E mutant thyroid cancer cell lines. Interestingly, embelin induced apoptosis in BRAF wild type cancer cell lines, even after treatment of SB203580. These results indicated that another signaling pathway, not a p38 pathway, might be existed for embelin induced apoptosis in BRAF wild type thyroid cancer cell. In conclusion, embelin-induced XIAP suppression resulted in an increase of apoptosis via phosphorylation of p38 in BRAF V600E-mutant thyroid cancer cell lines. Regulation of XIAP activity may be potentially useful as a treatment of thyroid cancer, especially in BRAF V600E-mutant thyroid cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1709. doi:1538-7445.AM2012-1709
Abstract 171: EGCG enhances the therapeutic potential of gemcitabine and CP690550 (Tasocitinib) by inhibiting STAT3 signaling pathway in human pancreatic cancer cells
Authors: Tang Su-Ni , Fu Junsheng , Shankar Sharmila , Srivastava Rakesh K.
Published: August 2012

Signal Transducer and Activator of Transcription 3 (STAT3) is an oncogene, which promotes cell survival, proliferation, motility and progression in cancer cells. Targeting STAT3 signaling may lead to the development of novel therapeutic approaches for human cancers. Here, we examined the effects of epigallocathechin gallate (EGCG) on STAT3 signaling in pancreatic cancer cells, and assessed the therapeutic potential of EGCG with gemcitabine or JAK3 inhibitor CP690550 (Tasocitinib) for the treatment and/or prevention of pancreatic cancer. Cell viability and apoptosis were measured by XTT assay and TUNEL staining, respectively. Gene and protein expressions were measured by qRT-PCR and Western blot analysis, respectively. The results revealed that EGCG inhibited the expression of phospho and total JAK3 and STAT3, STAT3 transcription and activation, and the expression of STAT3-regulated genes, resulting in the inhibition of cell motility, migration and invasion, and the induction of caspase-3 and PARP cleavage. The inhibition of STAT3 enhanced the inhibitory effects of EGCG on cell motility and viability. Additionally, gemcitabine and CP690550 alone inhibited STAT3 target genes and synergized with EGCG to inhibit cell viability and induce apoptosis in pancreatic cancer cells. Overall, these results suggest that EGCG suppresses the growth, invasion and migration of pancreatic cancer cells, and induces apoptosis by interfering with the STAT3 signaling pathway. Moreover, it can be combined with other chemotherapeutic agents like gemcitabine and CP690550 for the treatment and/or prevention of pancreatic cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 171. doi:1538-7445.AM2012-171
Abstract 1716: FGFR1 gene amplification predicts the antitumor response of FGFR inhibitor LY2874455 in human lung cancer cells
Authors: Bi Chen , Zhao Genshi , Baker Jessica A. , Starling James J. , Wu Wenjuan
Published: August 2012

Lung cancer is the leading cause of cancer-related death for both men and women in the world. FGFR receptor tyrosine kinase signaling pathway has been implicated in tumor cell growth, survival, angiogenesis, and metastasis. Activation of the FGFR pathway via FGFR over-expression, and chromosomal amplification and translocation as well as genetic mutation contributes to the tumor progression and the resistance to anti-EGFR and -VEGF treatment in lung cancer. Targeting FGFR pathway is therefore an attractive therapeutic strategy for lung cancer. We have discovered and developed a novel, ATP competitive and orally active small molecule inhibitor of FGFR, LY2874455, currently in clinic development. LY2874455 is a potent inhibitor against four FGFRs, and demonstrates a broad-spectrum of anti-tumor activity in a number of tumor xenograft models. In the current study, we evaluated the anti-tumor activity of LY2874455 in lung tumor cells with or without FGFR1 gene amplification. FGFR1 is known to be amplified in human lung cancer cells NCI-H1581, NCI-H520 and DMS-114, but not in NCI-H460, A549 and NCI-H1299. The mRNA expression levels of FGFR1 are shown to be higher in the FGFR1 amplified cells than those in the FGFR1 non-amplified cells. Consistent with this, the FGFR1 amplified cells exhibited the much more robust b-FGF-induced phosphorylation of FRS2 and ERK, which are also more sensitive to the inhibition by LY2874455 than that in the FGFR1 non-amplified cells. Furthermore, the FGFR1 amplified cells exhibit the better responses to the growth inhibition and apoptosis induction by LY2874455 than those in the FGFR1 non-amplified cells. Therefore, these findings suggest that FGFR1 plays an important role in lung cancer cell growth and survival, and that LY2874455 may provide a promising treatment option for some lung cancer patients whose tumor harbors FGFR1 gene amplification. The in vivo anti-tumor response of LY2874455 is being further investigated in the tumor xenograft models derived from these FGFR1 amplified or non-amplified human lung cancer cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1716. doi:1538-7445.AM2012-1716
Abstract 172: Utility of retinoid treatment in the reversion of the malignant phenotype of mammary tumors with alterations in the expression of protein kinase C (PKC) isoforms
Authors: Bessone Maria I. Diaz , Berardi Damian E. , Ciriglaino Stefano , Campodonico Paola B. , de Kier Joffe Elisa Bal , Todaro Laura B. , Urtreger Alejandro J.
Published: August 2012

Some of the most promising signaling pathways for breast cancer treatment include the PKC family, involved in proliferation and apoptosis, and the retinoid system mainly involved in differentiation. In this work we have developed a murine model of mammary adenocarcinoma-derived cells (LM3) that overexpress α or ≤ PKC isoforms in order to analyze whether elevated levels of these kinases alter cell sensitivity to retinoid treatment (ATRA). LM3 cells are poorly responsive to ATRA. The overexpression of PKCα induced alterations in LM3 in vitro behavior that could be associated with tumor progression. In this sense, PKCα increased proliferation and motility (wound coverage: 69.3±8.3% vs 41.2±5.1% in control cells p<0.05) of the transfected LM3 cells. Interestingly, at the same time PKCα sensitized LM3 cells to ATRA treatment as evidenced by a significant reduction in both parameters. Upon othotopic inoculation into syngeneic mice LM3-PKCα cells formed tumors with higher growth rate and metastatic potential than LM3-vector cells. In vivo treatment with an ATRA pellet reduced both the local tumor growth and the number of spontaneous lung metastases (Md [Range]: 9 [0-27] vs 55 [20-75] for LM3-PKCα treated or not respectively p<0.05). On the contrary PKC ≤ overexpression did not affect LM3 cells behavior either in vitro or in vivo. LM3 cells response to ATRA was not modulated by PKC∈ either. However, through a RARE-luciferase gene reporter assay, we could determine that the overexpression of PKC∈ increased the activity of the retinoic acid receptors. Our results show that PKCα overexpression induced a more aggressive phenotype but also conferred ATRA sensitivity to mammary tumor cells. On the other side high PKC∈ levels increased the activity of retinoid receptors but did not modulate biological cell responses to retinoids. We believe that the use of retinoids may be beneficial in patients with aggressive breast tumors associated with high PKCα expression levels.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 172. doi:1538-7445.AM2012-172
Abstract 1722: Prognostic markers in early stage non small-cell lung cancer (NSCLC)
Authors: Romkes Marjorie , Nukui Tomoko , Buch Shama , Pancoska Petr , Herazo-Maya Jose , Kaminski Naftali , Dacic Sanja , Weissfeld Joel L. , Bigbee William L. , Shapiro Steven D. , Siegfried Jill M.
Published: August 2012

Conventional clinical and pathologic risk factors in stage I non small-cell lung cancer (NSCLC) provide limited prognostic information. Novel prognostic biomarkers are needed to identify patients with highest recurrence risk who will receive the greatest absolute risk reduction from adjuvant chemotherapy/radiation. Transcriptome, epigenome, and global genetic variation analyses of resected NSCLC frozen tumor samples (n=81; 29 stage 1A/52 stage 1B) were performed in order to develop an integrated prognostic signature. Only tumors that were resected within 3 months of the first biopsy-proven diagnosis were used to reduce lead time bias in timing of removal of the primary tumor. Here, the association of gene expression with disease free survival (DSF) is described. Whole genome expression data were quantile-normalized using the Illumina GenomeStudio 2011.1 software. Log-transformed data were then processed in Biometric Research Branch (BRB)-Array Tools. Prognostic models were developed with the top ranking genes using the supervised principal component survival algorithm (PCA). Risk group membership was then assigned based on this multivariate model using leave-one-out cross validation and class prediction modeling. Both models adjusted for age, gender and tumor histology. For stage 1A cases, a 113 gene signature selected by fitting a Cox proportional hazard model (Cox) was able to discriminate between poor and good DFS (α=0.001). Upon further analysis using a class prediction algorithm support vector machine (SVM), we identified an 18 gene classifier that predicted outcome SN=0.952, SP=0.571). A 10-fold cross validation ROC curve reported a value of 0.748. For stage 1B cases, the Cox model identified 141 genes (α=0.01), but the SVM prediction algorithm showed reduced SN/SP (0.75/0.25) for a 12 gene classifier, indicative of possible increased heterogeneity among the 1B cases. Nine genes overlapped between the PCA and class prediction model gene signatures. These nine are members of common lung tumorigenesis pathways, i.e. G-protein signaling regulation (KRAS/BRAF/RASSFI); tyrosine protein kinase signaling (EGFR/ERBB2); nucleotide excision repair (ERRC5 and ERCC6) and genes implicated in cell differentiation regulated by miR200B (HGD) or miR338 (TXNRD1), miRNAs independently found to be associated with prognosis in our cohort. These initial results demonstrate that a gene expression profile can distinguish stage I NCSLC tumors and predict prognoses, but require further validation. Supported in part by NIH 5P50 CA090440, P30CA047904, UPMC Institutional Funds.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1722. doi:1538-7445.AM2012-1722
Abstract 1724: A gene expression profile test to resolve squamous cell carcinomas of head & neck from squamous cell carcinomas of the lung
Authors: Lal Anita , Panos Rebecca , Marjanovic Mira , Walker Michael , Fuentes Eloisa , Henner W. David , Buturovic Ljubomir , Halks-Miller Meredith
Published: August 2012

The differential diagnosis between squamous cell carcinomas of the head & neck and squamous cell carcinomas of the lung is often unresolved because the histologic appearance of these two tumor types is quite similar. We have developed and validated a gene expression profile test (Pathwork Tissue of Origin Head & Neck Test) that distinguishes head & neck squamous and lung squamous cancers in formalin-fixed, paraffin-embedded (FFPE) specimens using a 2160-gene classification model. The test was validated in a blinded study using a pre-specified algorithm and microarray data files for 76 metastatic, poorly differentiated or undifferentiated FFPE tumor specimens that had either a known head & neck squamous or lung squamous diagnosis. The study met the primary Bayesian statistical endpoint for acceptance. Measures of test performance include overall agreement with the known diagnosis of 82.9% (95% CI, 72.5% to 90.6%), an area under the ROC curve (AUC) of 0.91 and a diagnostics odds ratio (DOR) of 23.6. Head & neck squamous cancers (N=38) gave an agreement with the known diagnosis of 81.6% and lung squamous cancers (N=38) gave an agreement of 84.2%. Reproducibility in test results between three laboratories had a concordance of 91.7%. The Tissue of Origin Head & Neck Test can aid in resolving the important differential diagnostic question for head & neck and squamous cell carcinoma of the lung.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1724. doi:1538-7445.AM2012-1724
Abstract 1725: Molecular determinants of Sorafenib sensitivity in renal and non small cell lung - Patient derived tumors engrafted in nude mice
Authors: Vincent Vuaroqueaux , Foucault Frederic , Krumbach Rebekka , Schüler Julia , Amalou Zakia , Metz Thomas , Fiebig Heinz-Herbert
Published: August 2012

Introduction: Sorafenib (Nexavar(®)) is a potent multikinase inhibitor suppressing angiogenesis and promoting cell autophagy. Sorafenib has been shown to be active in a broad range of cancers models and is approved in the clinic for treatment of renal cell carcinoma and hepatocellular carcinoma. Sorafenib was shown to inhibit non small cell lung cancers (NSCLC), however a recent phase III study failed to demonstrate efficacy when given together with carboplatin and paclitaxel. Therefore, the identification of molecular determinants of Sorafenib sensitivity is of particular importance for better selecting NSCLC patients for treatment with this therapy. In the present study we used a large collection of NSCLC and renal cancer patient derived tumor models xenografted in nude mice (PDX) to compare Sorafenib anti-tumor activity in both cancer types and to identify molecular determinants of tumor sensitivity. Material and Methods: Sorafenib was tested in vivo in a panel of 33 different PDX including 8 renal and 25 NSCLC. The drug was given orally at 200 mg/kg/day for 12 days and tumor sensitivity was evaluated by tumor volume inhibition of the test group relative to the vehicle control group (T/C in %). Molecular characterization was done using targeted mutation analysis by Sequenom MassARRAY Oncocarta 3.0, Affymetrix SNP V6.0 array for gene copy number variation and HGU133 plus 2 arrays for transcriptome analysis. Results: The mean % T/C values were 42% in renal and 38% in NSCL PDX. Using a T/C value of 30% as cut-off for sensitivity, tumor response to Sorafenib were observed in 5/8 renal (62%) and in 8/25(32%) NSCL PDX. In NSCLC the response to Sorafenib was 2/5 (40%) in epidermoid, 2/7 (28%) in large cell and 3/13 (23%) in adeno subtypes. The transcriptome analysis revealed differentially expressed transcripts between tumors responding to Sorafenib and those being resistant (student t-test adjusted p<0.05). Gene ontology and pathway analyzes will be presented. In addition, for some of these transcripts, differential expression was associated with gene copy number variations in PDX. None of the specific gene mutations detected by Oncocarta panel. 3 were associated with Sorafenib sensitivity. Conclusion: Sorafenib is a potent tumor inhibitor not only in renal but also in the different NSCL cancer model subtypes. Molecular analysis evidenced differentially expressed transcripts but no gene mutation associated with response to treatment. The relevance of these markers for predicting Sorafenib sensitivity should be confirmed in an independent data set.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1725. doi:1538-7445.AM2012-1725
Abstract 1732: DNA methylation profiling defines clinically relevant biological subsets of non-small cell lung cancer
Authors: Walter Kim , Holcomb Thomas , Januario Tom , Du Pan , Hampton Garret M. , Amler Lukas C. , Yauch Robert L. , Bourgon Richard , Shames David S.
Published: August 2012

Purpose: Non-small cell lung cancers (NSCLC) comprise multiple distinct biological groups with different prognoses. For example, patients with epithelial-like (EL) tumors have a better prognosis and exhibit greater sensitivity to inhibitors of the epidermal growth factor receptor (EGFR) pathway than patients with mesenchymal-like (ML) tumors. Here we test the hypothesis that EL NSCLCs can be distinguished from ML NSCLCs on the basis of global DNA methylation patterns. Experimental Design: To determine whether phenotypic subsets of NSCLC can be defined based on their DNA methylation patterns, we combined microfluidics-based gene expression analysis and genome-wide methylation profiling. We derived robust classifiers for both gene expression and methylation in cell lines and tested these classifiers in surgically resected NSCLC tumors. We validate our approach using quantitative RT-PCR and methylation specific PCR in formalin-fixed biopsies from NSCLC patients who went on to fail front-line chemotherapy. Results: We show that patterns of methylation divide NSCLCs into EL and ML subsets as defined by gene expression and that these signatures are similarly correlated in NSCLC cell lines and tumors. We show that DNA methylation differences predict EL or ML phenotypes NSCLC cell lines, surgically resected tumors, and formalin-fixed biopsies from NSCLC patients who went on to fail front-line chemotherapy. Conclusions: Our data demonstrate that patterns of DNA methylation can divide non-small lung cancers into two phenotypically distinct subtypes of tumors and provide proof of principle that differences in DNA methylation can be used as a platform for predictive biomarker discovery and development.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1732. doi:1538-7445.AM2012-1732
Abstract 1733: Development of a novel blood plasma protein test for diagnosis of lung cancer
Authors: Streeper Robert T. , Izbicka Elzbieta , Diaz Armando , Campos David , Louden Christopher , Michalek Joel
Published: August 2012

Background: A blood test for detection of lung cancer (LC) could provide survival benefits in high-risk populations. We conducted a retrospective study as the next step of our effort to develop a test for diagnosis of non-small cell lung carcinoma (NSCLC), the most common form of LC, by measuring levels of protein biomarkers in plasma. Materials and Methods: We quantified levels of 106 proteinaceous biomarkers using multiplexed immunoassays of subjects a) diagnosed with stage I/II NSCLC, b) normal non-smoking controls, and c) normal heavy smoking controls (> 10 p.y.). All groups consisted of Caucasian men and women (93 subjects each, ages range 45-87, Ave 59). The specimens were randomized and tested in a blinded manner. A support vector machine (SVM) algorithm, developed and tested in a pilot study, was used to identify differentially expressed biomarkers in the disease and control groups taking into account patients’ gender. Pathway analysis was applied to characterize pathology- and gender-specific patterns of biomarker expression. Results: In a recent pilot study we developed SVM models that classified subjects to NSCLC or normal using 59 plasma markers or subsets thereof, for both genders combined or separately. When data for both genders were analyzed together, SVM classified subjects to NSCLC or normal and identified 4 differentially expressed markers: EGF, sCD40 ligand, IL-8 and MMP-8; sensitivity (SE) 0.93 (95% CI: 0.88-0.96), specificity (SP) 0.87 (95% CI: 0.80-0.92). Considering the sexes separately, the differentially expressed best subset of markers for men (EGF, IL-8, sFAS, MMP-9 and PAI-1) had both SE and SP of 1.0. For women, the best subset of markers (EGF, sCD40 ligand and IL-8) also yielded SE and SP of 1.0. The present study has further confirmed and refined our earlier findings and expanded panels of diagnostic biomarkers for NSCLC with comparable sensitivities and specificities. Analysis of cellular pathways represented by the discriminatory biomarker signatures suggested a role of sex hormone-driven differences of NSCLC plasma biomarkers. Conclusions: The set of diagnostic biomarkers for NSCLC identified in this study holds significant promise for use in the early diagnosis of lung cancer and warrants validation of findings in other ethnic groups and for potentially confounding diseases in clinical studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1733. doi:1538-7445.AM2012-1733
Abstract 1737: Co-existence of positive MET FISH status with EGFR mutations signifies poor prognosis in lung adenocarcinoma
Authors: Kobayashi Naomi , Aragane Naoko , Nakamura Tomomi , Sato Akemi , Takeda Yuji , Mitsuoka Masahiro , Yamasaki Fumio , Hayashi Shinichiro , Sueoka Eisaburo , Kimura Shinya
Published: August 2012

MET, which is located in 7q31, is a receptor tyrosine kinase for hepatocyte growth factor. Overexpression of MET has been reported in various malignancies and is associated with cell proliferation, inhibition of apoptosis, and metastasis. MET amplification has been reported to contribute to acquired resistance to epidermal growth factor tyrosine kinase inhibitors (EGFR-TKI) in non-small cell lung cancer(NSCLC), being observed among 5-10% of those patients. Therefore, MET gene alterations could be both prognostic and predictive. To evaluate MET gene copy number, fluorescence in situ hybridization (FISH), real-time PCR, and comparative genomic hybridization (CGH) have been used. The results of real-time PCR do not directly reflect cancer cells. CGH is not suitable for the evaluation of a large number of specimens due to cost. Therefore, FISH is the most practical method for evaluation of MET gene copy number. However, the frequency of positive cases varies due to a lack of standardized criteria. We evaluated MET gene copy number in lung adenocarcinoma and its association with clinicopathological characteristics. FISH was applied to evaluate high MET gene copy number and true amplification in 138 lung adenocarcinoma patients using two criteria: the Cappuzzo scoring system and PathVysion. MET positive cases according to the Cappuzzo scoring system evidenced both aneuploidy and true amplification, whereas PathVysion revealed only amplification. Proportion of MET FISH positive cases was 15% and 4% determined by the Cappuzzo system and PathVysion, respectively. PathVysion demonstrated higher frequencies of MET FISH positives among men and smokers and evidenced no MET FISH positives in patients with bronchioloalveolar carcinoma, which is recognized as pure lepidic growth without invasion of stroma, blood vessels, or pleura. Prognosis was significantly associated with MET FISH positive only as defined by the PathVysion system (gene amplification), not by the Cappuzzo system. However, progression-free survival time of patients with both EGFR mutations and MET FISH positive defined by the Cappuzzo scoring system was significantly shorter than with EGFR mutations alone. These results suggest that MET FISH is a potential prognostic factor and co-existence of EGFR mutation and MET FISH positive status evidences poorer prognosis compared to EGFR mutation alone, which is consistent with the results of in vitro experiments.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1737. doi:1538-7445.AM2012-1737
Abstract 174: Keratin 8 and 18 knockdown increases cisplatin-induced apoptosis and invasive potential through claudin1/PI3K/NFkB up-regulation in epithelial carcinoma cells
Authors: Fortier Anne-Marie , Cadrin Monique , Asselin Éric
Published: August 2012

Keratins are epithelial-specific intermediate filament (IF) proteins, which are expressed in a tissue-specific manner. As part of the cytoskeleton, keratins are important for the mechanical stability and integrity of epithelial cells and tissues. Moreover, a number of keratins are involved in intracellular signalling pathways which regulate response to injuries and non-mechanical stresses, cell growth, cell death and cancer progression. Keratins 8 and 18 (K8/18) are typically co-expressed as the primary keratin pair in simple epithelial cells and their expression are maintained during malignant transformation, hence their use as diagnostic marker in tumor pathology. However, in recent years different studies have shown that IF should not be considered only as markers proteins but also as regulators of cancer cell signaling and that they might play an active role in malignant transformation. In the present study, we addressed the question as to whether K8/18 expression affects tumor fate and behaviour. Our results show that K8/18 stable knockdown using shRNA increases cisplatin sensitivity in three different epithelial cancer cell lines. Indeed, western blot analysis of caspases activation and flow cytometry analysis of AnnexinV/PI staining show that K8/18 knockdown sensitizes cells to cisplatin-induce apoptosis. Increased FasL expression and FasR membrane targeting suggest that apoptosis is enhanced via the death receptor pathway. Moreover, using in vitro wound healing and transwell invasion assays, we observed that K8/18-knockdowned cancer cells display an increased cellular motility and invasion. Interestingly, we observed that these cells present higher PIP3 levels in the plasma membrane as determined by both fluorescence microscopy and flow cytometry analysis. Consequently, the K8/18-shRNA clones show PI3K/Akt/PKC/NFkB pathways hyperactivation and increased MMP-9 expression. Furthermore, these processes are shown to be partially regulated by the tight junction's protein claudin-1, which is highly increased in K8/18-shRNA clones. To our knowledge, these results represent the first indication that K8/18 can influence the phenotype of epithelial cancer cells. Knockdown of K8/18 increases cisplatin sensitivity and invasive potential of epithelial cancer cell lines through the regulation of different cell signaling pathways, involving claudin-1 dependent PI3K activation and NFkB transcription activity. These results support the hypothesis that modulation of K8/18 expression plays an active role in cancer progression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 174. doi:1538-7445.AM2012-174
Abstract 1756: Downregulation of topoisomerase IIα by all-trans retinoic acid in human cervical carcinoma cells
Authors: Li Xiaoting , Fu Shilong , Shen XIn , Zhou Donghu , Jiang Apei , Han Suping , Feng Qing
Published: August 2012

Cervical cancer remains a threatening disease to women worldwide. Topoisomerase IIα is a novel proliferation marker for cervical cancer diagnosis, therapy and prognosis. Understanding of the regulation of topoisomerase IIα would facilitate prevention and therapy of cervical cancer. In the present study, a significant repression of topoisomerase IIα level in stage Ib of squamous cervical carcinoma tissue was observed in patients with chemotherapeutic treatment, suggesting topoisomerase IIα is a target for chemotherapeutic drugs. All-trans-retinoic acid (ATRA), a chemopreventive agent, was shown to suppress both endogenous and exogenous topoisomerase IIα protein but not mRNA expression in human cervical carcinoma HeLa cells. Cycloheximide (CHX) prior to ATRA treatment augmented the topoisomerase IIα instability. ATRA induced the degradation of topoisomerase IIα via the proteasome pathway but not lysosome pathway. Ubiquitination was involved in this ATRA-induced topoisomerase IIα degradation. The ATRA-induced topoisomerase IIα repression occurred only in cell nucleus. Interestingly, overexpression of topoisomerase IIα enhanced cyclin D1 expression and caused HeLa cells de-differentiation while knockdown of topoisomerase IIα resulted in the cells differentiation and decreased cyclin D1. ATRA suppressed drug resistant protein P-gp while the typical topoisomerase II inhibitor etoposide did not. The molecular therapeutic implication of ATRA in cervical cancer therapy is discussed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1756. doi:1538-7445.AM2012-1756
Abstract 1761: Breast cancers with compromised DNA repair exhibit selective sensitivity to elesclomol-induced oxidative DNA damage
Authors: Alli Elizabeth , Ford James M.
Published: August 2012

Hereditary breast cancers due to germline mutations in BRCA1 and sporadic breast cancers of the basal-like subtype are often triple-negative, i.e. lack expression of estrogen and progesterone receptors and lack overexpression/amplification of the HER2/neu oncogene. Given that the triple-negative status of these cancers renders them relatively insensitive to existing “targeted” therapy, improved treatment options are needed. We tested a panel of cell lines that represent the different molecular subtypes of breast cancer for sensitivity to elesclomol, an experimental therapeutic that produces elevated levels of oxidative DNA damage, using the MTT assay. We found that both BRCA1-mutated and basal-like breast cancer cell lines with defective base-excision DNA repair (BER) were markedly more sensitive to elesclomol than cell lines that represent the normal breast or other subtypes of breast cancer with proficient BER. We also assessed sensitivity to elesclomol in breast cancer cell lines that were stably transduced with shRNA to OGG1, a BER glycosylase required for the repair of the most common type of oxidative DNA damage. Two different cell lines with decreased levels of OGG1 (shOGG1A, 50% knock-down; shOGG1B, 20% knock-down) were more sensitive to elesclomol (IC50 = 0.04nM and 0.3nM, respectively) compared to two different isogenic control cell lines (IC50 = 2nM and 3nM, respectively). Taken together, these data suggest that compromised repair of oxidative DNA damage by BER represents a functional target for elesclomol. Overall, BRCA1-mutated and/or basal-like breast cancers may benefit from treatment regimens that include elesclomol.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1761. doi:1538-7445.AM2012-1761