bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3925: IGF2 activation of HER2 receptor in HER2 positive breast cancer cell lines
Authors: Helu Xousaen M. , De Leon Daisy D.
Published: August 2012

Breast cancer is the second leading cause of death in women. Overexpression of Human Epidermal Growth Factor Receptor 2 (HER2) is detected in 20-30% of breast cancers. Overexpression of HER2 is linked to poor prognosis, more aggressive phenotype, and resistance to chemotherapy and hormonal treatment. HER2 is part of a four member family of cell surface receptors that are implicated in transmission of signals controlling cell growth and differentiation. HER2 has an intracellular domain with tyrosine kinase catalytic activity and dimerization of HER2 activates the MAP kinase and AKT pathways. HER2 receptor also heterodimerizes with the IGF1 receptor and can be activated by IGF1 receptor ligand. The insulin-like growth factor 2 (IGF2) has an important role in fetal and cancer development signaling through the insulin-like growth factor 1 receptor (IGF1R), insulin receptor (InsR), and cross-talk with the estrogen receptor alpha and beta (ERα, ERß). We hypothesized that binding of IGF-II to the IGF-1R/HER2 dimer may activate the HER2 signaling pathway. We characterized the IGF2 mRNA and protein levels in HER2 positive cell lines (BT474, SKBR3, and ZR 75-30) by qPCR and Western blot. We also analyzed the effect of IGF2 treatment on the phosphorylation levels of HER2. We measured the signaling levels of HER2 and IGF1R in the presence and absence of IGF2. Our study showed that IGF2 increases HER2 phosphorylation. Furthermore, blocking IGF1R increases HER2 protein levels. Thus, we propose that IGF2 has an important role in the activation of HER2 signaling pathway.

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 3925. doi:1538-7445.AM2012-3925
Abstract 3926: Abrogating tamoxifen resistance with the ribonucleoside analog, 8-amino adenosine
Authors: Stellrecht Christine M. , Dennison Jennifer B. , Schiff Rachel , Shentu Shujun , Gandhi Varsha
Published: August 2012

Hormone therapy is a prevalent treatment of choice for patients with estrogen receptor alpha (ERα) positive breast cancers. Unfortunately, due to resistance, many women do not benefit from these forms of therapy. AIB1 is a transcriptional co-activator that plays an important role in resistance to hormone therapy. Depending on the cell/gene context, agents such as tamoxifen either stimulate or antagonize the transcriptional activities of ERα. Tamoxifen binding alters ERα to an intermediate conformation with affinity for both co-activators and co-repressors, causing the conformation to be dependent on co-regulator levels in the cell milieu. The stoichiometric balance between AIB1 and co-repressors is a key determinant of tamoxifen resistance. Additionally, cross-talk between ERα and growth factor (GF) signaling further manipulates ERα activity and responsiveness to hormone therapy, particularly in cells with high AIB1 levels. Of the receptor signaling transducers, AKT, ERK, and p38 are key players in resistance. We hypothesized that strategies to target the stoichiometric balance of AIB1 to co-repressors would be clinically beneficial for abrogating tamoxifen resistance. Furthermore, it has been demonstrated that agents that are able to interfere with the ERα/GF crosstalk are also highly advantageous for abrogating resistance to tamoxifen. We have shown that the ribonucleoside analog, 8-aminoadenosine (8-NH2-Ado) is a transcription inhibitor. As such, it is expected to primarily deplete the expression of the small percentage (∼5%) of gene transcripts with short-half lives, such as AIB1. We have demonstrated that treatment of MCF-7 and BT-474 cells with non-tumoricidal concentrations of 8-NH2-Ado (1 µM) decreased AIB1 protein levels. Furthermore, using in vitro kinase assays, we demonstrated the active metabolite of 8-NH2-Ado is a selective inhibitor of MEK1, 2, and 6. An initial analysis of 4-hydroxytamoxifen dose response in two different tamoxifen resistant breast cancer cell lines, MCF-7/HER-2:18 and BT-474, demonstrated non-tumoricidal analog concentrations were able to reduce the dose of 4-hydroxytamoxifen needed to inhibit proliferation in these cells. Additionally, 8-NH2-Ado inhibited AKT and ERK1/2 activity in these cells. In conclusion, AIB1 and GF signaling molecules are attractive targets for reversing resistance to hormone therapy. Our results suggest that treating ERα positive breast cancer with combinations of tamoxifen and 8-NH2-Ado would be an effective strategy to overcome tamoxifen resistance.

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 3926. doi:1538-7445.AM2012-3926
Abstract 3928: Inhibition of the ubiquitin proteasome system differentially regulates glucocorticoid receptor-mediated transcriptional processes
Authors: Wiggins Kimberly R. , Davis Valerie , Phadke Dhiral , Shah Ruchir , Archer Trevor K.
Published: August 2012

The ubiquitin-proteasome system (UPS) plays an integral role in controlling protein turnover to modulate cellular processes such as growth, proliferation, differentiation, and apoptosis. This occurs through multiple mechanisms that rely on the regulation of numerous transcription factors such as steroid hormone nuclear receptors. The glucocorticoid receptor (GR) is a member of this receptor family and its activation promotes nuclear translocation and downstream transcriptional activity through the recruitment of co-regulatory complexes. Our goal is to understand the mechanisms by which the proteasome regulates glucocorticoid receptor-mediated gene transcription. To explore the effects of proteasome inhibition upon GR-mediated transcription, we used ChIP-Seq and microarray studies using breast cancer cells to examine GR binding and gene expression profiles after treatment with dexamethasone (dex) or dex in combination with a proteasome inhibitor, MG132 (dex + mg), for 1 hour and 18 hours. More than 14000 and 5000 GR-binding sites were apparent after both treatments in the 1 hour and 18 hour samples, respectively. GR binding sites seen during 1 hour of treatment can be mapped to more than 9000 genes while the binding sites in the 18 hour treatment samples mapped to over 5000 genes. In addition, over 55% of binding sites in all treatment groups are within 50 kilobases of annotated transcription start sites. To verify these putative interactions, we assayed the transcript levels of genes at both time periods and examined the levels of bound GR throughout the gene. The results reveal a differential regulation of genes that can be divided into three classes: dex-responsive, dex + mg-responsive, or dually responsive. Further analyses will reveal if there are clearly defined chromatin states that demarcate each class. These data provide insights into the intricate mechanisms of proteasome-mediated GR regulation and highlight proteasomal-regulated genes involved in numerous diseases and developmental disorders.

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 3928. doi:1538-7445.AM2012-3928
Abstract 393: Low-dose gamma-irradiation inhibits IL-6 secretion from human lung fibroblasts to suppress bronchial epithelial cell transformation by cigarette smoke carcinogen
Authors: Chen Wenshu , Xu Xiuling , Bai Lang , Padilla Mabel T. , Scott Bobby R. , Belinsky Steven A. , Lin Yong
Published: August 2012

The relationship between low-dose (<100 mSv) ionizing radiation (LDR) and human cancer risk is controversial. While microenvironment consisting of stromal cells, cytokines and growth factors is modulated by carcinogens to favor carcinogenesis, how LDR impacts this oncogenic process is not well understood. In this study, we investigated the role and mechanism of low-dose ionizing radiation in modulating the response of human lung fibroblasts to carcinogen stimulation and the impacts of which on bronchial epithelial cell transformation. Using a human cytokine antibody array, we found that secretion of proinflammatory cytokines IL-6, CXCL1 and CXCL5 from human lung fibroblasts was induced by cigarette smoke carcinogen benzo[a]pyrene diol epoxide (BPDE), which was inhibited by a low dose of γ-ray irradiation. BPDE induced IL-6 secretion from lung fibroblasts in a dose-dependent manner, which was suppressed when the NF-κB and ERK pathways were blocked. LDR effectively inhibited NF-κB but not ERK activated by BPDE, suggesting that NF-κB is the target for LDR in fibroblasts. In addition, conditioned media from BPDE-treated fibroblasts activated STAT3 in the immortalized human bronchial epithelial cell line Beas-2B, which was blocked with an IL-6 neutralizing antibody as well as by pretreating fibroblasts with LDR. Furthermore, IL-6 significantly enhanced BPDE-induced Beas-2B cell transformation in vitro. These results suggest that LDR inhibits cigarette smoke carcinogen-induced lung carcinogenesis through suppressing secretion of protumorigenic cytokines such as IL-6 from fibroblasts in the tumor-prone microenvironment.

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 393. doi:1538-7445.AM2012-393
Abstract 3931: Progesterone and estrogen affect the expression and secretion of heparin binding epidermal growth factor like growth factor (HB-EGF) in RL95-2 cells, an endometrial cancer cell line
Authors: Otenyo Phane Gesare , Williams Ebonee , Macaulay Daniel , Carson Laura , Regisford E. Gloria C.
Published: August 2012

Endometrial cancer is one of the most common gynecological cancers. Heparin Binding Growth Factor-like Growth Factor exerts acute effects on cancer cells; in that it induces and promotes proliferation and hence the pathogenesis and progression of endometrial cancer. Steroid hormones directly or indirectly affect the expression of HB-EGF and may thus play a role in the malignant transformation of endometrial cells. The aim of this project is therefore to evaluate the effects of progesterone (P4) and estrogen (E2) treatments on the expression and secretion of HB-EGF in endometrial cancer cells. We hypothesized that P4 will reduce HB-EGF expression and secretion while E2 will increase its expression and secretion. To test our hypothesis, an endometrial cell line, RL95-2, was treated for four days asfollows: (1) 10−8M E2, (2)10−6M P4, (3) 10−8M E2 + 10−6M P4 (4) 10−8M E2 in two days followed by 10−6M P4 and (5) Control. Immunocytochemistry (ICC), ELISA and western blot analysis were performed to evaluate the secretion and expression of HB-EGF in RL95-2 cells. Western blot analysis and ICC showed a consistently enhanced expression of HB-EGF in cells treated with E2 and in cells treated with E2 followed by P4 relative to the control groups. Alternatively, cells treated with P4 showed a down-regulation of HB-EGF expression relative to the control group. ELISA analysis showed high secretion of HB-EGF in cells treated with E2 and cells treated with E2 followed by P4 relative to the control groups. The secretion of HB-EGF was lower in cells treated with P4 alone compared to the control. In sum, there is an acute effect of E2 and P4 on the expression and secretion of HB-EGF and thus consequently may affect endometrial cancer cell proliferation.

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 3931. doi:1538-7445.AM2012-3931
Abstract 3932: IL13Rα2 regulates cell invasion, and is a therapeutic target for adrenocortical carcinoma
Authors: Jain Meenu , Zhang Lisa , Joshi Bharat , Puri Raj , He Mei , Kebebew Electron
Published: August 2012

Adrencortical neoplasms are one of the most common human neopolasms. Furthermore, it is difficult to distinguish localized benign from malignant adrenal tumors. Adrenocortical carcinoma (ACC) is a relatively rare but aggressive malignancy with limited therapeutic options for patients with advanced and metastatic disease. Several genome wide expression studies have shown overexpression of Interleukin-13 Receptor alpha2 (IL13Rα2) in some human malignancies but the function of this gene in cancer cells is poorly understood. We evaluated IL13Rα2 mRNA expression in normal (n=21), and in benign (n=78) and malignant (n=10) adrenocortical tumors. We found 11-fold higher expression in malignant tumors compared to benign (p<0.05) and absent expression in normal adrenocortical tissue. Immunohistochemistry analysis confirmed high protein expression in malignant and benign tumors and absent expression in normal adrenocortical tissues (p<0.05). IL13Rα2 mRNA expression level also had 90% accuracy for distinguishing benign from malignant adrenocortical tumors. We used IL13Rα2 siRNA knockdown to assess its effect on cell proliferation and cell invasion in the NCI H295R cell line, which expressed high levels of IL13Rα2. IL13Rα2 knockdown did not significantly affect cell proliferation but did decrease invasion by 60% as compared to negative control (p<0.05). To determine the significance of IL13Rα2 as a therapeutic target for ACC, we evaluated the sensitivity in two ACC cell lines (NCI-H295R with high IL13Rα2 expression and SW13 with low IL13Rα2 expression) to a highly specific IL-13 targeting immunotoxin (IL-13-PE). We found the IC50 for IL-13-PE was 1.3ng/ml (NCI-H295R) and 1000ng/ml (SW13) in the cell lines. Based on this observation, we then tested the efficacy of IL-13-PE therapy in an ACC xenograft model in athymic nude mice. Mice receiving either intratumoral or intraperitoneal IL-13-PE therapy as compared to control showed a significant reduction in tumor size as well as tumor necrosis (p<0.05), and had prolonged survival (p<0.05). Both routes of IL-13-PE administration were well tolerated with no significant toxicity. Our study indicates that IL13Rα2 is overexpressed in 100% of ACC, is a good diagnostic marker for ACC, regulates ACC cell invasion, and is a novel therapeutic target for the treatment of ACC.

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 3932. doi:1538-7445.AM2012-3932
Abstract 3934: HCV core protein upregulates the expression of vascular endothelial growth factor via the nuclear factor-α/hypoxia-inducible factor-1α axis under hypoxic conditions
Authors: Abe Mitsuhiko , Koga Hironori , Yoshida Takafumi , Masuda Hiroshi , Iwamoto Hideki , Sakata Masahiro , Hanada Shinichiro , Nakamura Toru , Taniguchi Eitaro , Kawaguchi Takumi , Yano Hirohisa , Torimura Takuji , Ueno Takato , Sata Michio
Published: August 2012

Aim: Hepatitis C virus (HCV) core protein critically contributes to hepatocarcinogenesis, which is often observed in liver cirrhosis. Since the liver cirrhosis microenvironment is affected by hypoxia, we focused on the possible driving force of HCV core protein on signal relay from hypoxia-inducible factor (HIF)-1α to vascular endothelial growth factor (VEGF). Methods: Human hepatocellular carcinoma cells stably overexpressing HCV core (Core cells) and NS5A (NS5A cells) were established; empty vector-transfected (EV) cells were used as controls. Hypoxia was induced by oxygen deprivation or by using cobalt chloride (CoCl2). YC-1 was used to inhibit HIF-1α expression. Protein analyses for cultured cells and liver tissues obtained from CoCl2-treated HCV core-transgenic (Core-Tg) mice were performed by Western blot and/or immunocytochemistry. Cellular mRNA levels were evaluated by quantitative real-time RT-PCR. Results: Under hypoxia, the sustained expression of HIF-1α, but not HIF-2α, was profoundly observed in Core cells but, was faint in EV and NS5A cells. Immunocytochemistry revealed increased HIF-1α in the nucleus. HIF-1α mRNA levels were significantly higher in Core cells than in EV cells under both normoxia and hypoxia. The HIF-1α-targeted VEGF and Bcl-xL expressions were increased in Core cells under hypoxia and abolished by YC-1 treatment. Hypoxic liver samples of Core-Tg mice indicated significant increases in both HIF-1α and VEGF expression compared to the wild type. Conclusions: HCV core protein has the distinct potential to transcriptionally upregulate and sustain HIF-1α expression under hypoxia, thereby contributing to increased VEGF expression, a key regulator in the hypoxic milieu of liver cirrhosis.

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 3934. doi:1538-7445.AM2012-3934
Abstract 3940: Activation of Signal Transducer and Activator of Transcription 3 (STAT3) in ovarian cancer: The hypoxic implications
Authors: McCann Georgia A. , Naidu Shan , Tierney Brent J. , Sudhakar Meryl , Cohn David E. , Selvendiran Karuppaiyah
Published: August 2012

Background: The hypoxic microenvironment plays a key role in tumorigenesis and the development of therapeutic resistance in epithelial ovarian cancer (EOC). The goal of this study was to examine the expression of STAT3 in hypoxic EOC, to understand its mechanism of activation, and to explore the therapeutic efficacy of a novel compound HO-3867. Methods: Expression of STAT family proteins in cell lines and human tumor specimens was analyzed via western blot and IHC. MTT was used to assess cell viability and apoptotic analysis was performed using flow cytometry. Over-expression or suppression experiments were performed using S1PR1 cDNA and shRNA. Tumor xenografts were made with cell lines exposed to hypoxic preconditioning. Results: pSTAT3 Tyr705 is highly expressed in ovarian cancer cell lines exposed to hypoxia as well as in hypoxic regions of advanced human EOC tumor specimens. Accelerated xenograft tumor growth was observed when implanted cells were pre-conditioned in a hypoxic microenvironment. Similar to our in vitro findings, the tumor xenografts of hypoxic ovarian cancer cells (HOCCs) had high expression of pSTAT3 and its target proteins. We found an increased expression of sphingosine-1-phosphate receptor 1(S1PR1) in HOCCs, a known positive feedback regulator of STAT3 transcription. Suppression of S1PR1 expression with shRNA significantly reduced the expression of pSTAT3 Tyr705 through the inhibition of the JAK kinases, resulting in decreased cell survival. Conversely, overexpression of S1PR1 with cDNA increased pSTAT3 Tyr705 and promoted cell proliferation under hypoxic conditions. Finally, treatment of HOCCs with the novel synthetic compound HO-3867 resulted in decreased expression of pSTAT3 Tyr705, as well as STAT3-regulated proteins VEGF, Bcl-xL, Bcl-2, and cyclin D1. The rapid downregulation of pSTAT3 Tyr705 was associated with accelerated ubiquitin-dependent degradation. An induction of apoptosis contributed to inhibition of cell proliferation as HO-3867-exposed cells exhibited elevated levels of cleaved caspases and PARP. Conclusions: The hypoxic microenvironment results in STAT3 activation as evidenced in our in vitro and in vivo models. S1PR1 is involved in STAT3 Tyr705 activation in hypoxic ovarian cancer via the JAK signaling pathway. The specific targeting of hypoxia-induced tyrosine phosphorylation of STAT3 with HO-3867 is a novel therapeutic strategy in the treatment of hypoxia-mediated chemotherapeutic resistant ovarian 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 3940. doi:1538-7445.AM2012-3940
Abstract 3945: HIFs regulate alternative splicing of HIF target genes
Authors: Sena Johnny , Wang Liyi , Hu Cheng-jun
Published: August 2012

Alternative splicing is the process in which identical pre-mRNA molecules are spliced in different ways, allowing individual gene to express multiple mRNAs or isoforms that encode proteins with diverse and even antagonistic functions. Alternative splicing occurs in over 70% of human genes, is important in normal development and many diseases including cancer. It is well known that most solid-tumors are hypoxic and the hypoxic microenvironment is a driving force for tumor progression and metastasis by hypoxia-inducible-factor (HIF)-mediated transcriptional programs that increase genes involved in tumor growth, angiogenesis, and metastasis. Here, we report a novel finding that transcriptional factor HIF not only activates gene expression, also regulates RNA splicing of its target genes, generating isoforms that favor tumor growth. Using an Exon Microarray we found that approximately 1000 genes are predicted to undergo hypoxia-mediated AS in Hep3B cells, 100 of those genes are hypoxia inducible genes. Using RT-PCR, RT-qPCR and WB we validated that the first 10 HIF target genes, CA9, ANGPTL4, WNK1, PLOD2, ADM, DUOX2, ENO2, P4HA2, EGLN3, and PDK1 undergo hypoxia-mediate AS, as predicted by exon array. Interestingly, AS of these genes is regulated by HIFs, not by hypoxia since: 1) siRNA knockdown of ARNT, HIF1, or HIF2 inhibits hypoxic induction of HIF target genes as well as inhibits hypoxia-mediated splicing changes; and 2) Normoxic HIF activity by deleting VHL, inhibiting PHD, or expressing normoxic-stable HIF protein led to hypoxic splicing patterns of HIF target genes, demonstrating HIF activity is sufficient to induce alternative splicing of HIF target genes. However, AS of HIF target genes cannot be explained by increased transcription of these genes since no splicing changes were observed when these genes were activated by hybrid protein with HIF transactivation domains replaced by VP16, E2F1, or SP1 transactivation domain. Further experiments indicated that the N-terminal transactivation domains (N-TAD) of HIF1α or HIF2α are responsible for splicing changes of these HIF-target genes. In summary, we have uncovered a novel function of HIFs in the regulation of target gene splicing. The discovery of HIF's role in mRNA splicing of its target genes will change the dogma concerning the role of HIF in cancer progression. We propose that HIFs promote cancer progression not only through transcription activation but also through alteration of mRNA splicing of HIF targets. RO1 CA134687 NIH/NCI (Hu)

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 3945. doi:1538-7445.AM2012-3945
Abstract 3948: Proteome analysis of ovarian cancer cell lines under hypoxic conditon by two-dimensional difference gel electrophoresis (2D-DIGE)
Authors: Kikuchi Ryoko , Iwaya Kei-ichi , Matsubara Osamu
Published: August 2012

The cancer cells which survived hypoxia are known to have aggressive properties and can easily metastasize to other regions, however, the molecular basis of hypoxia-induced changes of cancer cells is not clear. We used a proteomic approach to identify the hypoxia-responsive proteins in 3 ovarian cancer cell lines, KFr13, RMUG-S and ES-2. The cell lines were incubated under lower oxygen concentration (5%) and under normal oxygen concentration (20%) in 48 hours. Even in lower oxygen concentration, the cells survived and increased in number in these 48 hours. The total proteins from the cells were extracted and analysed by 2D-DIGE and MALDI-TOF-MS. We identified the protein, HRP1, whose molecular weight is 71KDa, that all 3 cell lines showed decreased expression under lower oxygen condition compared with normal condition. HRP1 secretion is a growth arrest signal for rat mammary adenocarcionma cells. HRP1 is one of the new candidates for hypoxia-related proteins in ovarian 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 3948. doi:1538-7445.AM2012-3948
Abstract 3953: Senescence induced of smooth muscel protein 22-alpha (SM22) and Radiation patients change human-link genes expression level
Authors: Kim Myeong Ok , Kim Younjoo , Ko Younmi
Published: August 2012

P53 and RB protein is famous for its capacity to suppress tumor senescence pathway member. Senescence is a leading cause of cancer-related death, considerable uncertainty still remains about their effectiveness. However, each marker in an increased risk that experience genomic data with a cancer-specific mutations. So, senescence marker that techniques can't develop patient-specific treatment for particular about experimental study-conditions and result-analysis. This study was undertaken to evaluate the sm22 (smooth muscel protein 22-alpha) protein relation of senescence marker and changes condition of the radiation at human-link senescence marker genes. We were determined the expression of senescence-associated β-galactosidase and western-blot assay. It was performed in colon cancers that were 17 patients each of normal and adenoma, 16 patients each of normal and adenocarcinoma. To conformed enzyme efficiency is β-galactosidase, senescence pathway using of p16, p21 and p53. On the other hand, sm22 use of senescence marker in study. we have decided to micro-array analysis that 3 patients each of normal and adenoma, 2 patients each of normal and adenocarcinoma, 2 patients each of normal and irradiation-adenocarcinoma (2 weeks after) available miRNA sequences on the Agilent Human whole genome 8x60K array chip. Gene expression levels were calculated with Feature Extraction v10.7.3.1 (Agilent technologies, CA) Relative signal intensities for each gene were generated using the Robust Multi-Array Average algorithm. Senescence induced p16 expression in dependent β-galactosidase assay results, sm22 protein suppressed cellular senescence levels as well as p16 protein. The expression of p16 and sm22 was analyzed that not involved each normals, adenomas and adenocarcinomas tissue by western blot analysis. In addition, it found that these results were similar to those of micro array. The three groups (adenoma, adeno-carcinoma and irradiation-adeno-carcinoma) can be compared in terms of total 2-fold changed gene (least 200% of controls for up-regulated and lower than 50% of controls for down-regulated genes). We found this integrative data analysis tool to the LCN2, LYPD5 and LOC100132051 genes as well as CDKN2 (p16) and TAGLN2 (sm22) gene expression. Irradiation groups was much less than the adenoma (45.3 %) and adenocarcinoma (71.1 %) 2-fold changed gene expression. The LCN2, LYPD5 and LOC100132051 genes lower than adeno-carcinoma group level. Our observations suggest that shared gene and protein expression profiles may contribute to sm22 and p16 of senescence marker, irradiation induces cellular senescence of colon 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 3953. doi:1538-7445.AM2012-3953
Abstract 3954: mTOR inhibitor increases radiation sensitivity in a radiation-resistance cancer cells through induction of cellular senescence
Authors: Nam Hae Yun , Lee Yoon Sun , Lee Hyang Ju , Kwak Jihye , Jung Ji Young , Kim Seong Who , Kim Sang Yoon
Published: August 2012

Autophagy is frequently activated in cancer cells resistant to ionizing radiation and is considered to limit the efficacy of radiotherapy as a prosurvival mechanism. In autophagy signaling, mTOR is an important target for enhancing the therapeutic effect of radiation since inhibiting mTOR limits cell growth and decreases cell proliferation, while increasing autophagy. However, the mode of these actions has remained poorly understood. Here, using radioresistant carcinoma cells, we investigated the mechanism by which the enhanced autophagy flux through the mTOR inhibition overcomes radioresistance of cancer cells subjected to irradiation. AMC-HN-9 cells triggered robust cellular proliferation, and induced cell arrest and autophagy activation in an irradiation dose-dependent manner, showing that adoption of an autophagy mode serves as a survival strategy against radiation. Interestingly, however, the potentiation of radiation-induced autophagy by concomitant inhibition of mTOR activity with mTOR inhibitors (rapamycin or PP242) reduced rather than enhanced cell survival and cell proliferation. Notably, mTOR inhibition in radiation-exposed cells was accompanied by senescence features, including the increased senescence-associated β-galactosidase (SA α-gal) activity and heterochromatin formation and irreversible growth arrest. Stress-induced senescence was associated with suppression of Rb-mediated E2F target genes. The results of this study show that the mTOR-dependent induction of autophagy can maintain a growth-arrested state in radioresistant cancer cells and increase radiosensitivity. It indicates that cellular senescence induced by mTOR inhibitor is a promising interventional strategy for enhancing the therapeutic efficacy in cancer cells refractory to irradiation

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 3954. doi:1538-7445.AM2012-3954
Abstract 3955: Overexpression of the 58-kDa microspherule protein (MSP58) triggers cellular senescence via the p53-p21 signaling axis
Authors: Hsu Che-Chia
Published: August 2012

The nucleolar protein 58-kDa microspherule protein (MSP58) is a candidate oncogene implicated in modulating cellular proliferation. In our studies, acute depletion of MSP58 expression using short hairpin (sh)RNA synthesis led to aneuploidy and apoptosis, whereas ectopic expression of MSP58 regulated cell proliferation in a context-dependent manner. Remarkably, ectopic expression of MSP58 in HT1080, NIH3T3, normal human fibroblasts, and mammary epithelial cell lines resulted in induction of premature senescence, caused large and flattened cell morphology, and increased senescence-associated β-galactosidase activity. This phenotype was accompanied by hypophosphorylated Rb, elevation of p53 and p21 protein expressions, activation of p53-mediated transcription, accumulation of phosphorylated forms of ATM/ATR and Chk1/Chk2 kinases, and formation of γH2AX and RPA70 foci, indicating that MSP58 activates the DNA damage response (DDR) and p53/p21 signaling pathways. Furthermore, we identified the SWI/SNF chromatin remodeling subunit, BRG1, as a critical mediator of MSP58-induced senescence. MSP58, BRG1, and p53 form a ternary complex on the p21 promoter, and collaborate to activate p21. This study revealed novel links among MSP58, p53, and cellular senescence in human tumors and normal 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 3955. doi:1538-7445.AM2012-3955
Abstract 3958: Multi-Reg: An integrative parallel approach to uncover drivers of cancer
Authors: Akavia Uri David , Danussi Carla , Sanchez-Garcia Felix , Iavarone Antonio , Pe'er Dana
Published: August 2012

Tumor samples harbor a vast number of genomic alterations of various kinds, and it is difficult to distinguish driver alterations that contribute to oncogenesis from passenger alterations. Most computational methods identify driver genes by focusing on the most frequent copy number alterations. Previous work in our lab has led to the development of CONEXIC, a Bayesian algorithm that identifies candidate driver genes in cancer and links them to gene expression signatures they govern by integrating copy number and gene expression (Akavia et al, Cell, 2010). This algorithm was applied to data from melanoma cell lines, where it correctly identified known drivers, such as MITF and KLF6, and connected them to their known targets. In addition, it predicted novel tumor dependencies not previously implicated in melanoma, which were validated experimentally. Drivers may act concurrently, where not only the strongest one is important. For example, either PTEN deletion or AKT activation can lead to a similar expression signature and phenotype. Therefore, we developed a new algorithm, based on the same principles as CONEXIC, with multiple improvements. The new algorithm - Multi-Reg - is capable of detecting multiple candidate regulators that can all act in parallel to regulate an expression signature, and is also capable of integrating mutations in addition to copy number and gene expression data. We applied Multi-Reg to glioblastoma data from The Cancer Genome Atlas (TCGA). This data includes copy number, gene expression and mutations for hundreds of primary tumor samples. Multi-Reg has identified putative drivers that were missed by CONEXIC, such as the known genes PDGFRA and NF1, in addition to EGFR & ERB2 (identified by CONEXIC). Additionally, since Multi-Reg candidate drivers act in parallel, we can group them by shared targets. For example, EGFR & ERRB2 induce the same genes, which represent the Mesenchymal subtype of glibolastoma. These same genes are repressed by PDGFRA & NF1. This matches the known behavior of glioblastoma subtypes, where EGFR & PDGFRA characterize the Mesenchymal and Proneural subtypes, respectively. Thus, our results correctly identify known drivers of glioblastoma. Additionally, Multi-Reg results identified RHPN2 as a novel oncogenic factor controlling a gene expression signature related to adhesion. Validation has shown that while RHPN2 has no effect on cell proliferation it induces invasiveness in glioblastoma cell lines. This shows that Multi-Reg cannot only discover drivers, but can link them to the oncogenic phenomena they controls, and suggest the appropriate biological validation for it.

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 3958. doi:1538-7445.AM2012-3958
Abstract 3961: Computational modeling of pancreatic cancer reveals growth and dissemination kinetics and suggests optimum treatment strategies
Authors: Haeno Hiroshi , Gonen Mithat , Davis Meghan B. , Herman Joseph M. , Iacobuzio-Donahue Christine A. , Michor Franziska
Published: August 2012

Pancreatic cancer is a leading cause of cancer-related death, largely due to metastatic dissemination. We investigated pancreatic cancer progression by utilizing a mathematical framework of metastasis formation together with comprehensive data of 228 patients, 101 of whom had autopsies. We found that pancreatic cancer growth is initially exponential. After estimating the rates of pancreatic cancer growth and dissemination, we determined that patients likely harbor metastases at diagnosis and predicted the number and size distribution of metastases as well as patient survival. These findings were validated in an independent database. Finally, we analyzed the effects of different treatment modalities, finding that therapies which efficiently reduce the growth rate of cells earlier in the course of treatment appear to be superior to upfront tumor resection. These predictions can be validated in the clinic. Our interdisciplinary approach provides insights into the dynamics of pancreatic cancer metastasis and identifies optimum therapeutic interventions.

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 3961. doi:1538-7445.AM2012-3961
Abstract 3962: Mathematical models for cellular pharmacodynamics of fractionated radiation and radiochemotherapy
Authors: El-Kareh Ardith W. , Jones Leslie Braziel , Secomb Timothy W.
Published: August 2012

Cellular response to radiation is generally described by the linear-quadratic equation, but the extension to complicated fractionated schedules, and to combined exposure with chemotherapeutic drugs is not well-established. Mathematical models for cellular response are needed for predictive quantitative modeling of treatment. Here, such models are formulated using the previously developed concepts of peak damage models and additive damage, which in prior studies proved successful in describing cellular pharmacodynamics of single-agent and combination chemotherapy. The exponential form of the linear-quadratic equation is replaced by Hill or cumulative lognormal functions of the peak value over time of a quantity termed “cellular damage.” Damage is a saturating function of radiation dose. Cell survival depends on whether the peak value of cellular damage ever exceeds a threshold value during treatment. This model is tested for a range of data sets for single-dose radiation, and compares favorably with the linear-quadratic. In one data set (Park et al 2008) the dose range was sufficiently wide to show the significant overestimation of cell kill by the linear-quadratic model at high doses; the Hill/lognormal formulation behaves more satisfactorily in this limit. The model is then extended to the case of fractionated radiation through the assumption that damage decays between treatments according to first-order kinetics, and damage from different cycles is additive. The model gives a satisfactory fit to literature data with multiple fractionation schedules for the same drug and cell line. Finally, the principle of additive damage is applied to model cellular response to radiochemotherapy, with damage as a linear superposition of terms from radiation and drug. Overall, the peak cellular damage model with additive damage was found to fit in vitro dose-response data well, and provides a tool for understanding and predicting response as schedule and dose are varied in combination treatments. It is expected that these models will have application in assessment of preclinical cellular pharmacodynamics data, as well as computational simulations of treatment response.

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 3962. doi:1538-7445.AM2012-3962
Abstract 3964: Inference of robust tumor phylogenetic markers from multi-sample data
Authors: Subramanian Ayshwarya , Schwartz Russell , Shackney Stanley
Published: August 2012

Computational cancer phylogenetics can play an important role in delineating possible tumor progression pathways and identifying molecular subtypes and mechanisms of action. We previously developed a pipeline for constructing tumor phylogenies from recurring cell types computationally inferred from whole genome copy number data. The accuracy and detail of these tumor phylogenies, however, depends on the identification of accurate and high-resolution molecular markers of progression, i.e., reproducible regions of copy number variation that can be used to robustly differentiate different subtypes and stages of progression. Here we present a new method for the problem using hidden Markov models (HMMs) to derive robust, high resolution progression markers from sets of tumor samples. We demonstrate our method on a publicly available array comparative genome hybridization (aCGH) dataset (NCBI GEO GSE16672, Navin et al., 2010) from sectioned primary ductal breast tumors. Our method uses an HMM, a class of probabilistic models, to classify sets of aCGH data into a partitioning of samples into normal (diploid) or amplified at each copy number probe. It differs from other similar HMM methods primarily in seeking a parsimonious set of combinations of amplification states able to explain all aCGH profiles simultaneously in order to identify robust markers of progression across samples. The model learns frequencies with which different combinations of amplifications are observed across the samples by modeling individual probes as Gaussian random variables with either normal or tetraploid means, with data more consistent with tetraploid being classified as amplified and those more consistent with diploid classified as normal. To handle a combinatorial explosion in combinations of amplification states with increasing numbers of samples, the method introduces a Gibbs sampling algorithm to learn a parsimonious model of the most frequently occurring combinations of amplification states. We applied our methods to a previously constructed set of inferred cell types derived from the Navin et al. data (Tolliver et al., 2010) and to a comparison set of 9 random samples from the raw aCGH data. We validated our model relative to manual labeling of amplicons on the same data. In both experiments, the HMM method was able to pick up significantly larger numbers of robustly amplified segments per chromosome than did prior methods or manual analysis. The resulting segments can be directly fed into downstream analysis routines for phylogeny inference or other predictions. In future work, the HMM method may be improved by fine-tuning the underlying model for copy number variation.

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 3964. doi:1538-7445.AM2012-3964
Abstract 425: Targeting embryonic signaling pathways in melanoma
Authors: OReilly Kathryn E. , Poliseno Laura , de Miera Eleazar Vega-Saenz , Friedman Erica , Hernando Eva , Osman Iman
Published: August 2012

Melanoma incidence is quickly rising and the average survival for a patient with metastatic disease is less than one year. Novel targeted therapies, such as B-Raf mutant kinase inhibitors, have shown dramatic initial responses, but resistance to single agent therapy inevitably ensues. We hypothesize that melanoma stem-like cells may be important in melanoma progression and may play a role in chemoresistance to targeted therapy. We have profiled melanoma cell lines as well as metastatic melanoma tissue for activation of embryonic stem cell pathways via microarray and quantitative RT-PCR. We observed upregulation of the Hedgehog pathway in a subset of cell lines and in human melanoma tissue and found that Smoothened (SMO) expression was associated with significantly shorter recurrence-free survival in metastatic melanoma patients. We also found that small molecule inhibition or genetic silencing of Smoothened (SMO) decreased melanoma cell proliferation and migration in vitro and identified overexpression of GLI2 as a marker of resistance to SMO inhibition. These findings suggest that inhibitors of hedgehog pathway signaling, already in clinical trials in other tumor types, may be a novel strategy for advanced melanoma.

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 425. doi:1538-7445.AM2012-425
Abstract 4252: Activated ALK collaborates with MYCN in neuroblastoma pathogenesis
Authors: Zhu Shizhen , Lee Jeong-Soo , Guo Feng , Shin Jimann , Perez-Atayde Antonio R. , Kutok Jeffery L. , Rodig Scott J. , Neuberg Donna S. , Helman Daniel , Feng Hui , Stewart Rodney A. , Wang Wenchao , George Rani E. , Kanki John , Look A Thomas
Published: August 2012

Neuroblastoma is a developmental tumor that arises in the peripheral sympathetic nervous system and accounts for 10% of all cancer-related deaths in children. The anaplastic lymphoma kinase (ALK) gene is mutationally activated in a subset of primary neuroblastomas, including those with MYCN gene amplification, suggesting pathogenic cooperation. Because the mechanism underlying this cooperation is unclear, we generated a novel transgenic zebrafish model that overexpresses human MYCN and activated ALK in the peripheral sympathetic nervous system to analyze their interaction. The expression of MYCN in this model induces neuroblastomas in the inter-renal gland, the zebrafish analogue of the adrenal medulla, which is the site of origin observed in approximately half of childhood neuroblastomas. Furthermore, the tumors resemble human neuroblastomas histologically, immunohistochemically and ultrastructurally. Concomitant expression of activated ALK with MYCN in this model profoundly accelerates the onset of neuroblastoma and markedly increases disease penetrance. Detailed in vivo analyses show that MYCN overexpression induces adrenal sympathetic neuroblast hyperplasia, blocks chromaffin cell differentiation, and triggers a developmentally-timed apoptotic response in the hyperplastic sympathoadrenal cells. Coexpression of activated ALK with MYCN provides prosurvival signals that block this apoptotic response and allow continued expansion and oncogenic transformation of hyperplastic neuroblasts. Taken together, these findings provide a mechanism for the synergistic interplay of MYCN with activated ALK in neuroblastoma pathogenesis and demonstrate the utility of the zebrafish model in understanding human disease processes that may aid the development of improved therapeutic strategies.

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 4252. doi:1538-7445.AM2012-4252
Abstract 4256: Use of short tandem repeat profiling for authentication of human cell lines
Authors: Reid Thomas M. , Wilson Keen , Kraemer Christopher , Baird Michael
Published: August 2012

Human cell lines are widely used in biological and biomedical research, and accurate data interpretation depends on unambiguous identity of a cell line relative to its original source. Contamination and misidentification of human cell cultures has been identified as a long-standing issue within the scientific community, and steps are now being taken to address the problem. Short-tandem-repeat (STR) DNA profiling provides an accurate, reliable, and standardized method for authentication of human identity. Currently, the ATCC is considering proposed standards for cell line authentication utilizing human STR typing, and many journals (including all AACR published journals) have adopted the NIH-recommended policy to mandate cell line authentication as a requirement for publication. Here we demonstrate the validation and use of two different STR systems to authenticate human cell lines as well as detect contamination by mouse feeder cells and other human cell lines. Threshold levels for detection of complete profiles as well as contamination are compared between the systems, and typical typing issues specific to cell lines are 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 4256. doi:1538-7445.AM2012-4256