bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4994: Downstream targets of Id1 transcription factor in non-small cell lung cancer
Authors: Nair Sajitha G. , Chellappan Srikumar
Published: June 2012

Id1 (Inhibitor of DNA binding/Differentiation) is a helix loop helix (HLH) protein, regulates transcription by binding to bHLH transcription factors and preventing their binding to the DNA. Id1 is especially known to mediate repression of E box proteins and Ets transcription factors. Id1 gene is overexpressed in lung, breast, pancreatic and prostate cancer and it is thought to promote the progression and metastasis of these tumors. At the same time, not much is known about its role in non-small cell lung carcinoma.Preliminary studies from our lab shows that Nicotine as well as EGF induces Id1 in both adenocarcinoma and squamous carcinoma cells and is notably upregulated in metastatic lung cancers. Depletion of Id1 also prevents the proliferation and invasion of NSCLS cells. Attempts were made to understand the downstream effectors of Id1 function in lung cancer progression and metastasis. As a step in this direction, A549 (K-Ras mutant, EGFR wild-type) and H1650 (K-Ras wild-type, EGFR mutant) cells were transfected with short interfering RNA (siRNA) for Id1and stimulated with 1μm Nicotine or 100ng/ml EGF. The samples were subjected to microarray analysis to identify various genes upregulated and downregulated in the Id1 siRNA transfected cell lines. 200 genes were upregulated 2 fold or more by Nicotine and 150 by EGF. Few genes such as stathmin like-3 (STMN3), GSPT1 (G1 to S phase transition), TPD52 (tumor protein D52) were downregulated in the absence of Id1 suggesting those are Id1 regulated genes. The microarray results were validated using Real-time PCR for the above genes. Further depletion of these three genes resulted in decreased cell proliferation, migration and invasion. Indeed STMN3 promoter could be induced by Id1 and Id1 was necessary for Nicotine to induce this gene. These results might reveal the mechanisms by which Id1 promotes tumor progression and metastasis. STMN3 is a microtubule destabilizing phosphoprotein which is overexpressed in adenocarcinoma as well as squamous cell carcinoma and induced tumor cell proliferation, migration and matrix invasion in respective cell lines. GSPT1, which is also known as eukaryotic translation release factor eRF3 and TPD52 are also reported to be over expressed in various kinds of cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4994. doi:10.1158/1538-7445.AM2011-4994
Abstract 4997: The isolation of miRNA targeting EGFR gene in lung cancer
Authors: Yamaguchi Gaku , Ikeda Norihiko , Kuroda Masahiko , Ohira Tatsuo , Uchida Osamu , Kajiwara Naohiro , Usuda Jitsuo , Kakihana Masatoshi , Saji Hisashi , Takanashi Masakatsu , Fujita Kohji , Yoshida Kohichi , Kataba Hiroaki , Iwasaki Kentaro , Tanaka Masami
Published: June 2012

MicroRNAs (miRNAs) belong to a class of the endogenously expressed non-coding small RNAs which primarily function as gene regulators. Growing evidence suggests that microRNAs have a significant role in tumor development.

Lung cancer, predominantly non-small cell lung cancer (NSCLC), remains the leading cause of cancer related deaths worldwide. Although EGFR signaling is important and well studied with respect to NSCLC progression, little is known about how miRNAs mediate EGFR signaling to modulate tumorigenesis.

In this study, we identified miRNAs that directly regulate the EGFR gene expression. We employed computational tools such as Pictar and Target Scan, and putative targets of 88 miRNAs were identified within the EGFR mRNA. Then, we analyzed the expression of EGFR in miRNAs-transfected human lung cancer cells (H3255, A549, Hcc827) by real time-PCR methods. In addition, we performed argonaute (AGO) IP experiments to ascertain whether these algorithmically predicted miRNA targets are biologically true targets in EGFR. Convincingly, we found that two miRNAs directly regulated the EGFR and both of these miRNAs also inhibited cell growth in cell viability assays. Identifying microRNA regulators of EGFR may contribute in development of novel therapeutics, and non-invasive early detection biomarkers for NSCLC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4997. doi:10.1158/1538-7445.AM2011-4997
Abstract 4999: FK506 binding protein 51 (FKBP51) sustains stemness and the metastatic potential of malignant melanoma
Authors: Romano Simona , Sorrentino Antonio , Pace AnnaLaura Di , Staibano Stefania , Mascolo Massimo , Bisogni Rita , Ilardi Gennaro , Romano Maria Fiammetta
Published: June 2012

Background. FKBP51 is a cochaperone with peptidyl-prolyl isomerase activity that regulates several biological processes in the cell, through protein-protein interaction. There is increasing evidence that FKBP51 hyperexpression is associated with cancer and this protein has a relevant role in sustaining cell growth, malignancy, and resistance to therapy. Very recently, we have found that FKBP51 is a protein associated with malignant melanoma. Its expression correlates with tumor aggressiveness and is maximal in metastatic lesions. Moreover, we have found that melanoma cancer stem cells express FKBP51 at levels higher than the other tumor cells and observed a 4 to 40 fold increase in the number of cancer stem cells, in cultures of melanoma cells stably transfected with FKBP51. Mitotic index, stemness features and angiogenesis are the main signs of aggressive melanoma. To better understand the role of FKBP51 in the biology and progression of melanoma, we attempted to investigate if this protein regulated the expression of molecules associated with tumor progression, namely the Chromatin Assembly Factor-1 (CAF-1) p60; the ATP-Binding Cassette family members ABCG2 and ABCB5; and the pro-angiogenic factor, endoglin or TβRIII. Methods. The melanoma cell line SAN was used for these experiments. FKBP51 knock down was obtained with siRNA, FKBP51 hyperexpression was obtained by stably transfecting SAN with a vector containing the FKBP51 human gene under a CMV promoter. Protein expression was measured by western blot and flow cytometry; mRNA was quantified by Realtime PCR. Results. We found that ABCG2 and CAF1 p60 mRNA levels were reduced by 5 and 2.5 folds, respectively, in FKBP51-silenced cells, in comparison with non-silenced cells. Reduced ABCB5 and CAF-1 p60 protein levels were detected by Western blot. By contrast, a clear increase of ABCB5 and CAF-1 p60 protein levels was observed in FKBP51 hyperexpressing cells. Flow cytometry showed that 39% of melanoma cells expressed endoglin. Such percentage was decreased to 5% by FKBP51 silencing, whereas it increased up to 78% in FKBP51 hyperexpressing cells. Finally, we found that CAF-1 and endoglin were co-expressed in more than 80% of ABCG2+ cells, whereas their expression was <40% in ABCG2- cells. Conclusions. FKBP51 regulates molecules involved in chromatin rearrangement, stemness and metastatization of malignant melanoma suggesting that this cochaperone may act as a transcriptional coactivator of oncogenes in melanoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4999. doi:10.1158/1538-7445.AM2011-4999
Abstract 5: Synthesis and biological evaluation of 5, 5’ substituted apogossypol and apogossypolone derivatives as pan-active inhibitors of anti-apoptotic B-cell lymphoma/leukemia-2 (Bcl-2) family proteins
Authors: Wei Jun , Placzek William J. , Stebbins John L. , Reed John C. , Pellecchia Maurizio , Fisher Paul B. , Dash Rupesh , Jamieson Catriona , Kitada Shinichi
Published: June 2012

Overexpression of anti-apoptotic Bcl-2 family proteins is commonly related with tumor maintenance, progression, and chemoresistance. Inhibition of these anti-apoptotic proteins is an attractive approach for cancer therapy. Guided by nuclear magnetic resonance (NMR) binding assays and computational docking studies, a series of 5, 5’ substituted Apogossypol and Apogossypolone derivatives was synthesized and identified pan-active antagonists of anti-apoptotic Bcl-2 family proteins, with binding potency in the low micromolar to nanomolar range. Apogossypol derivative BI97C1 inhibits the binding of BH3 peptides to Bcl-XL, Bcl-2 and Mcl-1 with IC50 values of 0.31, 0.32 and 0.20 µM, respectively. In a cellular assay, BI97C1 potently inhibits cell growth in the H460 human lung cancer and PC-3 human prostate cancer cell lines with EC50 values of 0.40 and 0.13 µM, respectively, and effectively induces apoptosis of the BP3 human lymphoma cell line (EC50 = 0.049 µM) in a dose-dependent manner. Apogossypolone derivative BI97D1 also potently inhibit cell growth in many cancer cell lines. BI97C1 and BI97D1 further display in vivo efficacy in transgenic mice in which Bcl-2 is overexpressed in splenic B-cells and also demonstrated superior single-agent antitumor efficacy in mouse xenograft model and represent promising drug leads for the development of novel apoptosis-based therapies for cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5. doi:10.1158/1538-7445.AM2011-5
Abstract 5003: β-catenin regulates multiple oncogenes in glioma
Authors: Yue Xiao , Han Lei , Lan Fengming , Yang Yang , Shi Zhendong , Zou Jian , Pu Peiyu , Kang Chunsheng
Published: June 2012

Gliomas are the most common primary intracranial tumors. Understanding the molecular basis of gliomas progression is required to develop more effective therapies. Although, disregulated of Wnt/β-catenin pathway is a hallmark of many cancers, little is known about its role in human gliomas. The expression of components of Wnt pathway in 80 astrocytic glioma specimens with different grades was examined by immunohistochemistry. The protein expression levels of Wnt1, Wnt2, β-catenin and Cyclin D1 were positively correlated with World Health Organization (WHO) grades of gliomas. To explore whether β-catenin can serve as a therapeutic target for glioma, we knockdowned β-catenin expression by siRNA and found that apoptosis was induced, cell proliferation and invasive ability were inhibited in vitro in U87, LN229 and A172 cells; xenograft tumors from β-catenin siRNA-treated LN229 cells were suppressed in vivo. To investigate the underlying mechanisms, we extracted gene/protein interactions by text mining Pubmed abstracts. Screening Pubmed by natural language processing, we identified 543 genes that interacted with β-catenin. Integrating multiple data sources through the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), we created the β-catenin related genes knowledge-driven network, identifying and interactions of this network with EGFR pathway. In both in vitro and in vivo studies, our results confirmed down-regulation of β-catenin induced reduced expression of EGFR, STAT3 and AKT1 mRNA and protein, besides, the level of phosphorylated Akt also decreased. A similar reduction in expression of CyclinD1, MMP2 and MMP9, downstream genes of the EGFR pathway, was observed. We examined the promoter region of EGFR and STAT3 gene for regulating DNA-binding elements and revealed the presence of putative β-catenin/TCF-4-binding sites. Further, EGFR and STAT3 were identified as direct targets of the Wnt/β-catenin pathway in glioma cells by ChIP and luciferase reporter assay. Taken together, we demonstrate that Wnt/β-catenin pathway is overexpressed in glioma and correlats closely with the progression of gliomas and EGFR and STAT3 are regulated by β-catenin/TCF-4 at the transcriptional level and interruption of β-catenin supresses the EGFR pathway by blocking multiple oncogenic targets in human glioma cells. Wnt/β-catenin pathway is of critical importance in the gliomagenesis and intervention of this pathway may provide a new therapeutic approach for malignant gliomas.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5003. doi:10.1158/1538-7445.AM2011-5003
Abstract 5006: Interleukin-1 receptor type II regulates matrix metalloproteinases expression and plays oncogenic roles in HaCaT cells
Authors: Lee Hui-Ju , Mar Ai-Chung , Lee Te-Chang
Published: June 2012

Interleukin-1 receptor type II (IL1R2) is an IL-1 decoy receptor, which plays a role to inhibit exogenous IL-1 signaling. Intriguingly, IL1R2 also performs as a pro- inflammatory factor via activation of intracellular IL-1α signaling in a variety of cells. In our previous study, we reported that long term arsenic exposure resulted in increased IL1R2 expression which led to increased motility of human urothelial cells. In the present study, we similarly found that the expression of IL1R2 was increased in long-term arsenic exposed HaCaT cells (A1, A2) and the tumorigenic derivative, T4R2 cells. Since matrix metalloproteinases (MMPs) are frequently increased in a variety of malignant tissues including skin cancer, we also revealed that a positive association of IL1R2 and MMP1 and MMP7 at either mRNA or protein levels, which were analyzed by quantitative real time RT-PCR and western blotting, respectively. To reveal the involvement of IL1R2 in up-regulation of MMPs, we adopted shRNA technique to silence the expression of IL1R2 in T4R2 cells. Our results demonstrated that MMP1 and MMP7 were suppressed in IL1R2 silencing T4R2 cells. Meanwhile, we found the enhanced expression of MMP1 and MMP7 in HaCaT cells with ectopically expressed IL1R2. Furthermore, we confirmed our previous finding showing that IL1R2 overexpression was accompanied with increased expression of precursor form of IL-1α protein (preIL-1α). Immunofluorescence staining showed the colocalization of IL1R2 and preIL-1α in the nucleus. In a xenograft mouse model, we found that HaCaT cells with ectopical expression of IL1R2 formed tumor in nude mice whereas IL1R2 silencing in T4R2 cells suppressed the tumorigenicity. Taken together, these results implicate that IL1R2 plays an oncogenic role in human keratinocyte transformation by upregulation of MMPs expression.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5006. doi:10.1158/1538-7445.AM2011-5006
Abstract 5009: The expression pattern of claudin-1, claudin-3, claudin-4, claudin-5 and claudin-7 in penile squamous cell carcinoma
Authors: Bulgarelli Adriana , Lourenço Silvia V. , Coutinho-Camillo Cláudia M. , Buim Marcilei EC , Begnami Maria Dirlei F.S. , Soares Fernando A. , Vassallo José
Published: June 2012

Claudins are a family of proteins that are structural and functional components of tight junctions and have crucial roles in the maintenance of cellular arrangement, adhesion and paracellular transport. Recent studies have shown that changes and/or loss of claudin expression plays an important role in tumorigenesis and tumor progression, and altered expression of claudins has been reported in various human carcinomas. Penile squamous cell carcinoma (SCC) is a major health problem in Brazil, corresponding to 2,1% of all brazilian men tumors and 5-year survival rate drops dramatically in patients with lymph node metastasis. Purpose: The aim of this study was to investigate claudins expression profile in penile SCC. Method: We analyzed expressions of claudin-1, -3, -4, -5 and -7 in 14 cases of penile SCC by relative gene expression method using real time PCR. Results: The expression of claudin-1, -4 and -5 was downregulated 15.9-, 6.9- and 9.1-fold, respectively, in 72% of penile SCC cases in comparison to normal penile squamous cells (p=0.0052 for both claudin-1 and -4 and p=0.0002 for claudin-5). Claudin-3 expression was variable, with 7 of the 14 cases (50%) showing low expression of claudin-3. Claudin-7 was upregulated 101.4-fold in 10 of 14 cases analyzed (p=0.067). Conclusion: Our data suggests that loss of claudin-1,-4 and -5 expression and gain of claudin-7 expression might be associated with the tumorigenic process in penile SCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5009. doi:10.1158/1538-7445.AM2011-5009
Abstract 501: Characterization of lung cancer stem/initiating cells from non-small-cell lung cancer patients
Authors: Chen Wan-Jiun , Chen Huei-Wen , Ho Chao-Chi , Chen Hsuan-Yu , Yang Pan-Chyr
Published: June 2012

Cancer stem cells (CSCs) have been proposed to be responsible for chemoresistant, tumor recurrent, and metastasis. To culture and characterize the CSCs become urgent and may be helpful to develop novel diagnostic and therapeutic strategies. Accumulated evidences indicated the importance of CSCs in tumor formation and successfully isolated the CSCs from patients or cell lines with different cancer types; however, this has not been well explored in lung cancer. Not alike previous isolation through specific markers and quiescent condition, we cultured the lung cancer stem/initiating cells from patient with lung adenocarcinoma and the CSCs formed tumor sphere as fed by the surrounding stroma cells. These lung cancer stem/initiating cells were characterized with highly expressed levels of the stemness makers, nanog, oct4, sox2, and klf4 (53.3-, 37.7-, 13.0, and 4.4-folds changes) as comparing to the feeder cells, also, showed relative higher percentages of side population (27.4%), ALDH activity (14%), higher tumorigenesity compared to other lung cancer cell lines including adenocarcinoma (A549, H522, H23, Hop62, and EKVX), large cell carcinoma (Hop92), squamous cell carcinoma (H226), and bronchi alveolar carcinoma (H322M). These lung CSCs also showed higher drug resistance capacity represented by IC50 to chemotherapy drugs including docetaxel, cisplatin, vinorelbine distrate, and etoposide (13.0-, 1.55, 27.2- and 103.3-folds increased). Furthermore, we found that as less as 100 cells could generate adenocarcinoma formation both in orthotropic or subcutaneous models in severe combined immunodeficient (NOD/SCID) mice. Interestingly, the lung CSCs could spontaneous differentiate into epithelial types of cancer cells as removing the feeder cells in vitro or different phenotypes of adenocarcinoma cells in vivo. Transcriptomic analysis indicated that the Wnt, Notch, TGF-β and HGF signaling pathways are highly expressed in the lung CSCs and quickly down-regulated as removing the feeder cells. The gene expression profiles also provide the novel markers for selection CSCs from lung cancer cell lines and lung cancer patients. To sum up, we have cultured the cancer stem/initiating cells from lung cancer patient with the feeder cells, which providing the tumor microenvironment to maintain the stemness of the cancer stem cells. This model should be helpful not only for isolating and cultured the lung CSCs from patients, but also in developing the anti-cancer strategy targeting on lung CSCs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 501. doi:10.1158/1538-7445.AM2011-501
Abstract 5011: Difference in expression of STAT3 and STAT5b in Swedish and Egyptian patients with activated b-cell subtype of diffuse large b-cell lymphoma
Authors: Jacobson Therese , Bahnassy Abeer A. , Lundahl Joachim , Tamm Katja Prokovskaja , Porwit Anna , Zekri Abdel-Rahman N. , Khaled Hussein M. , Mellstedt Håkan , Moshfegh Ali
Published: June 2012

Diffuse Large B-cell lymphoma (DLBCL), the most common type of lymphoma in adults, accounts for approximately 30 to 40% of newly diagnosed lymphomas. Using gene expression profiling, DLBCL can be divided into three immunologically indistinguishable subtypes: the germinal center B-cell like (GBC), the activated B-cell like (ABC) and the primary mediastinal B-cell lymphoma (PMBL) subtype. The aim of the study was to compare gene expression profiles of Swedish and Egyptian ABC DLBCL patients.

Formalin fixed and paraffin embedded material of 67 Egyptian and 15 Swedish ABC DLBCL patients were used in the study. Affymetrix gene expression platform was used for profiling and the analysis was carried out using GeneSpring GX software. Quantitative PCR (qPCR) and confocal microscopy was used to verify the findings.

The GeneSpring GX analysis showed that 3247 genes were at least 2 fold up- or downregulated comparing the two patient groups. Among these were 1827 genes upregulated and 1420 genes downregulated. Gene expression analysis was verified using qPCR showing that STAT3 was upregulated with a fold change of 3.1 in the Swedish patients and that STAT5b was upregulated with a fold change of 1.3 in the Egyptian patients. Confocal microscopy showed protein activity of STAT3 and 5b in the nucleus of Swedish and Egyptian patients respectively using monoclonal antibodies.

This is the first study that reveals diversity in expression of STATs in different populations of ABC DLBCL patients. STAT3 and 5b are known to have different pathways of activation and targeted disruption has shown their functional differences. Using gene expression profiling, we revealed different expression levels of STAT3 and STAT5b in Swedish and Egyptian ABC DLBCL patients. Since the populations of Sweden and Egypt are exposed to different infectious agents, the results of the present study further raise the question concerning viral etiology in ABC-DLBCL possibly due to repeated infections and chronic inflammation. In general, these findings give additional evidence to the importance of signaling pathways involving STATs in ABC DLBCL.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5011. doi:10.1158/1538-7445.AM2011-5011
Abstract 5018: Adherence to free breast cancer screening among uninsured black and Hispanic women
Authors: Okoronkwo Nneoma O. , Onojobi Gladys , Burnside Clinton , Williams Carla , McDonald-Pinkett Shelly , Frederick Wayne A.I. , Laiyemo Adeyinka O.
Published: June 2012

Background: Studies have shown that blacks have a higher mortality rate from breast cancer. However, it is unclear if this can be mainly attributed to their different biological susceptibilities or lower health care access or lack of adherence to scheduled procedures such as screenings and treatments or a combination of everything.

Aim: We sought to examine the extent to which uninsured black and Hispanic women adhere to free breast cancer screening appointments.

Methods: This study included a cohort of 410 participants from a monthly program in Howard University Cancer Center that offers free breast cancer screening (clinical breast examination and mammogram) for uninsured women who are aged 40 and older in the Washington DC area. These participants were referred from community health organizations in Maryland, District of Columbia and Maryland. Data was extracted from July 2009 to June 2010. The participants were categorized as non-adherent (no shows) if they did not cancel their appointments and did not show up for the breast cancer screening. We used Poisson regression models to evaluate the association of race with adherence to the scheduled breast cancer screening and adjusted for potential confounders. We calculated Relative Risks (RR) for no shows and 95% Confidence Intervals (CI).

Results: There were 195 (47.6%) Hispanic and 215 (52.4%) black women. A total of 314 (76.6%) women showed up for screening but 96 (23.4%) were non-adherent. When compared with Hispanic women, black women were older (mean age = 50.7 years vs 48.4 years; P = 0.0005), and more likely to be born in the United States (29% vs 2%; P < 0.0001) but were less likely to be married (31% vs 49%; P < 0.0001). A total of 42 (21.6%) Hispanic and 54 (25.1%) black women did not show up for the breast cancer screening, adjusted RR = 1.06: 95% CI: 0.71 – 1.57; P = 0.788.

Conclusions: There is a high rate of non-adherence to scheduled free breast cancer screening among underserved black and Hispanic women. Increased outreach effort and community education about this potentially preventable cancer is needed.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5018. doi:10.1158/1538-7445.AM2011-5018
Abstract 5023: Pretreatment blame, physical symptoms, and distress in patients undergoing radiotherapy for head and neck cancer
Authors: Ojo Bukola , Badr Hoda , Milbury Kathrin , Genden Eric M.
Published: June 2012

INTRODUCTION: Head and Neck Cancer (HNC) patients report higher rates of psychiatric morbidity than other cancer patients, suggesting that psychological adjustment is an important component of their quality of life. One factor contributing to patients’ distress is the degree to which they experience physical symptom morbidity. Given that HNC is often attributed to unhealthy lifestyle factors (i.e., tobacco, alcohol, and risky sexual behaviors resulting in HPV), another possible contributor, that has not been studied, is patients’ attributions of self-blame regarding their disease cause. The objective of this study was to explore the associations between lifestyle factors, symptoms, self-blame, and distress in this population. Understanding these potential interactions may provide targets for future psychosocial interventions.

METHODS: Data for this exploratory study was collected from a cross-sectional survey of 70 HNC patients (87 % male; 63% oropharyngeal) prior to the initiation of radiotherapy.

RESULTS: In terms of lifestyle factors, 70% of patients had a tobacco history and 7% had screening scores indicative of alcohol dependence as measured by the AUDIT; 61 % of the oropharyngeal patients were HPV positive. Behavioral self-blame was significantly greater among patients with a history of tobacco use (t(66) = 2.52, p = 0.01) and characterological self-blame was highest among patients with HPV positive tumors (t(66) = 2.28, p = 0.03). Although patient distress (BSI-18) was significantly associated with higher levels of cancer symptom severity (p = 0.0001) and interference (p = 0.0001), as measured by the MDASI-HN, the association of distress with blame also approached significance (p < 0.10).

CONCLUSION: Results suggest that blame is prevalent in head and neck cancer patients and is related to lifestyle factors that may have caused the disease. The finding that patient distress was associated with both symptom severity and self-blame is interesting and suggests that psychosocial interventions should not only address patients’ current level of symptoms but also their attributions of blame regarding the cause of their disease. Our future work will prospectively examine whether changes in symptoms and blame affect the course of distress in this vulnerable population.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5023. doi:10.1158/1538-7445.AM2011-5023
Abstract 5032: Targeting Src in mucinous ovarian carcinoma
Authors: Matsuo Koji , Nishimura Masato , Stone Rebbeca L. , Komurov Kakajan , Shahzad Mian MK , Bottsford-Miller Justin N. , Enomoto Takayuki , Kimura Tadashi , Ram Prahlad T. , Schmeler Kathleen M. , Frumovitz Michael , Gallick Gary G. , Wong Kwong K. , Sood Anil K.
Published: June 2012

BACKGROUND: Mucinous ovarian carcinomas have a distinct clinical pattern compared to other subtypes of ovarian carcinomas. We evaluated (i) stage-specific clinical significance of mucinous ovarian carcinomas in a large cohort and (ii) the functional role of src kinase in pre-clinical models of mucinous ovarian carcinoma.

METHODS: 1302 ovarian cancer patients including 122 (9.4%) cases of mucinous carcinomas were evaluated for survival analyses. Gene microarrays with src specific functional analysis were performed to identify biomarkers for 2 mucinous ovarian cancer cell lines compared with 5 serous ovarian cancer cell lines. Biological effects of src kinase inhibition were tested in a novel orthotopic mucinous ovarian cancer model (RMUG-S-ip2) using dasatinib-based therapy, an oral src kinase inhibitor.

RESULTS: Patients with advanced-stage (stage III/IV) mucinous ovarian cancer had significantly worse survival compared to those with serous histology: median overall survival, 1.67 versus 3.41 years, p=0.002; and median survival time after recurrence, 0.53 versus 1.66 years, p<0.0001. Gene expressions of mucinous ovarian cancer cell lines were distinct from serous ovarian cancer cell lines. Pathway analyses showed that src kinase might be a key factor in regulating a number of signaling pathways. Among 10 ovarian cancer cell lines, the RMUG-S-ip2 mucinous ovarian cancer cells showed the highest src kinase activity. Moreover, oxaliplatin treatment induced phosphorylation of src kinase and Akt. These induced activities were inhibited with concurrent administration of dasatinib. Combination therapy of oxaliplatin with dasatinib further demonstrated significant synergistic effects on reducing cell viability (interaction index <1), increasing apoptosis (mean percentage of total apoptotic cells in RUMG-S-ip2, untreated vs dasatinib alone vs oxaliplatin alone vs oxaliplatin and dasatinib, 4.1% vs 8.5% vs 9.5% vs 26.8%, p<0.01), and in vivo anti-tumor effects in the RMUG-S-ip2 model (tumor reduction rate in combination therapy: versus oxaliplatin monotherapy 79.1%, and versus dasatinib monotherapy 74.7%, both p<0.01).

CONCLUSION: Our results suggest that poor survival of women with mucinous ovarian carcinoma is associated with resistance to cytotoxic therapy. Targeting src kinase with combination of dasatinib and oxaliplatin may be an attractive approach for further clinical development in mucinous ovarian carcinoma.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5032. doi:10.1158/1538-7445.AM2011-5032
Abstract 5033: Stromal-derived conditioned media activates STAT3 via JAK2 and TYK2 and results in de novo resistance to Nilotinib
Authors: Nair Rajesh R. , Tolentino Joel H. , Argilagos Raul F. , Zhang Ling , Pinilla-Ibarz Javier , Hazlehurst Lori A.
Published: June 2012

Complete eradication of CML has not been possible in patients because of the presence of minimal residual disease (MRD) in sanctuary sites like the bone marrow (BM). One of the reasons for MRD is the milieu of cytokines and growth factors present in the BM microenvironment, that contribute to BCR-ABL independent drug resistance in CML. For example, in the present study, we demonstrate the importance of STAT3-mediated drug resistance in the BM microenvironment by showing that conditioned media (CM) derived from both normal and patient-derived mesenchymal stem cells and osteoblasts leads to STAT3 phosphorylation (pSTAT3-Y705) and concomitant drug resistance to Nilotinib in CML cells. Inhibiting the activity or reducing the expression of JAK2 and TYK2 was necessary for sensitizing cells to Nilotinib in CM. Moreover, increased pSTAT3-Y705 levels were associated with increased STAT3 transcriptional activity and up-regulation of Mcl-1 and cyclin D1. Our study indicates that there is an inherent redundancy in CM-mediated STAT3 activation pathway because concomitant neutralization of two potent activators of STAT3, IL-6 and G-CSF, does not reverse CM-mediated protection against Nilotinib. Furthermore, utilizing a SCID-hu bone implant in vivo model, we show that K562 cells with reduced STAT3 showed significantly lower tumor burden, which could be attributed to not only the slower proliferation rate, but also to the observation that these cells underwent cell death in BM stromal derived CM. Finally, CD34+ cells from CML patients when exposed to CM showed an increase in STAT3 phosphorylation which correlated with the development of de novo resistance to Nilotinib-mediated cell death. Taken together our data indicate that STAT3 is an important target in CML and its direct inhibition, along with BCR-ABL inhibitors, will likely be the most effective therapeutic strategy for reversing resistance within the BM microenvironment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5033. doi:10.1158/1538-7445.AM2011-5033
Abstract 5043: Norcantharidin induces melanoma cell apoptosis through TR3 pathway
Authors: Liu Shujing , Yu Hong , Kumar Suresh , Martin James , Xu Xiaowei
Published: June 2012

Norcantharidin (NCTD) is the demethylated analog of cantharidin, a naturally occurring toxin isolated from Mylabris. NCTD has been used to treat hepatocellular carcinoma with clinical efficacy and limited side effects. However, the underlying mechanism of its antitumor effects remains elusive. We have previously shown that TR3 expression is significantly decreased in metastatic melanomas and it is implicated in regulation of melanoma cell apoptosis. In this study, we showed that NCTD inhibited melanoma cell viability and induced apoptosis in vitro. NCTD induced translocation of TR3 from nucleus to mitochondria where it colocalized with Bcl-2 in melanoma cells. NCTD also increased Cytochome c release from mitochondria to the cytoplasm. There was also increased expression of Bax and cleaved caspase-3, accompanied with decreased expression of Bcl2 and Nf-kB2. The effects of NCTD were inhibited by knockdown TR3 expression in melanoma cells. To study its function in vivo, we treated Tyr::cre;BRAFCa/+;Ptenlox/lox mice with NCTD. Temozolomide and PBS were used as controls, and ten mice were used in each group. Temozolomide modestly increased mice survival, whereas NCTD significantly improved the survival of treated mice. Our data indicates that NCTD inhibits melanoma growth by inducing cell apoptosis via activation of a TR3 dependent pathway. These results suggest that NCTD is a potential therapeutic agent for melanoma.

Financial support was provided by CA-093372, CA-116103 to XX.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5043. doi:10.1158/1538-7445.AM2011-5043
Abstract 5047: Survival signal pathways as potential therapeutic targets in soft tissue sarcoma cell lines
Authors: Han Jung Woo , Kwon Bo Ram , Park Chan Hee , Lee Gui Youn , Jeung Hei-Cheul , Kim Se-Kyu , Roh Jae Kyung , Chung Hyun Cheol , Rha Sun Young
Published: June 2012

Introduction: Soft tissue sarcomas (STS) are rare malignancies but have poor survival rate. Surgery with standard chemotherapy has been the mainstay of treatment but newer strategies are necessary to improve survival. The investigations for targeted therapy are underway but little is known about potential targets until now. We previously presented potential molecular targets in osteosarcoma (OS) cell lines (101st AACR annual meeting 2010, abstract No.2451). We tried to compare the protein and mRNA profilings between the OS and STS cell lines to suggest the feasibility for developing molecular targeted agents in STS.

Methods: We examined the protein expression of c-Met, Akt, mTOR, and their phosphorylated status, and PTEN in seven OS (MG-63, HOS, KHOS/NP, SK-ES-1, U-2OS, Saos-2, and G-292) and five STS cell lines (SK-UT-1, A-673, SW-871, SW-982, HT-1080) using western blotting. The gene expression profilings of the cell lines were performed by 44K microarray (Agilent Inc.).

Results: Microarray data showed that whole genome expression pattern of STS was different from OS cell lines. But, the expressions of 94 mRNAs within the signal transduction pathways, which represent the currently utilized drug targets, revealed similar pattern between two groups of cell lines. To verify the possibility of developing molecular targeted agents in STS, we examined the activation patterns of signal transduction pathway proteins in each STS cell line.

All STS cell lines except A-673 with low c-Met, but high p-Met, expressed moderate level of c-Met and p-Met. PI3K was expressed in all cell lines, with higher p-PI3K in SK-UT-1 and A-673 than in other STS cell lines. All but SW-872 cell line expressed Akt, and SK-UT-1, SW-872 and HT-1080 had higher levels of p-Akt. SW872 showed negligible PTEN and high level of p-Akt. SW982 generally repressed level of p-Met, p-PI3K, p-Akt, p-mTOR comparing with other STS cell lines. Meanwhile, mTOR and p-mTOR were expressed relatively even in all STS cell lines.

Conclusion: We presented that STS cell lines had variously activated signal pathway status as in OS cell lines. Common potential targets such as mTOR was revealed, and the specific targets including PI3K or Akt were also presented. It suggests the possibility of developing specific molecular targeted agents in each type of STSs.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5047. doi:10.1158/1538-7445.AM2011-5047
Abstract 5049: Cyclopamine modulates ABCG2 activity in glioblastoma side population cells
Authors: Balbuena Jana , Pachon Gisela , Lopez-Torrents Guillermo , Fan Xing , Castresana Javier S. , Petriz Jordi
Published: June 2012

The Sonic Hedgehog pathway (HH) has been implicated in the maintenance of stem cells in adult tissues. Abnormal HH activation is also associated with initiation of neoplasia by cancer stem cells. Cyclopamine has been used as an inhibitor of the HH, with potential effectiveness in several xenograft models, including gliomas. Side Population (SP) cells are stem cells with expression of ABCG2, a multidrug ABC transporter, able to extrude dyes as Hoechst 33342 (Ho342). SP cells, then, present low fluorescence after staining with Ho342. ABC transporters have been identified as drug-resistance proteins. HH regulates the expression of ABC transporters, and promotes multi-drug resistance by increasing drug efflux by the ABC transporters. Different research groups have shown that SP cells can be depleted following cyclopamine treatment, an observation that would support that SP cells are HH pathway-dependent.

We hypothesized that cyclopamine might interact with ABCG2 and limit the number of SP cells. Therefore, we analyzed the effect of cyclopamine and temozolomide on the MXRA cell line (KB cells transfected with the full-length ABCG2 cDNA), and on five astrocytoma cell lines, both in the parental cell lines and in their SP cells. We first evaluated the modulatory effect of cyclopamine on ABCG2 function by measuring the intracellular accumulation of two substrates of ABCG2, Ho342 and mitoxantrone. Secondly, we analyzed the effect of these drugs in the phenotype of SP cells (the cells were incubated with only Ho342, or a combination of Ho342 and cyclopamine, or a combination of Ho342, cyclopamine and temozolomide; afterwards we analyzed the number of SP cells).

We saw that cyclopamine significantly decreased the fraction of SP cells but not through a depletion of the SP compartment, but as a consequence of the molecular mechanisms involved in the affinity of cyclopamine and Ho342 for ABCG2, as cyclopamine modulated the ABCG2 transporter by increasing Ho342 and mitoxantrone uptake. Temozolomide had no effect on the SP cells.

Therefore, cyclopamine, although decreases the SP fraction of cancer stem cells in glioblastoma cell lines, does not deplete the SP compartment, but modulates ABCG2 activity.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5049. doi:10.1158/1538-7445.AM2011-5049
Abstract 505: Multiscale mathematical model of skin reveals the critical role of stromal aging in melanoma initiation
Authors: Kim Eunjung , Basanta David , Rebecca Vito , Messina Jane , Mathew Rahel , Smalley Keiran S. , Anderson Alexander R. A.
Published: June 2012

Melanoma initiation is known to involve specific genetic changes as well as the disruption of both cell-cell and cell-microenvironment interactions. However, the mechanisms by which dysregulated cell-cell interactions lead to melanoma development remain elusive. To better understand melanoma initiation and progression we have developed a hybrid multiscale mathematical model of normal skin (virtual skin). The model focuses on key cellular and microenvironmental variables that regulate normal homeostatic skin function, and is constructed to recapitulate how keratinocytes and fibroblasts regulate melanocyte proliferation, migration and death.

Experimentally, we have shown that as fibroblasts age and become senescent their behavior changes significantly, leading to the expression of multiple growth factors, matrix proteins and proteases. We also showed, using 2D/3D culture, that interactions with senescent fibroblasts can aid melanoma initiation.

Using our virtual skin model, we systematically investigated the effects of disrupting interactions between melanocytes, keratinocytes, fibroblasts and the microenvironment in driving melanoma initiation. The resulting virtual skin pathology readily recapitulates a spectrum of aberrant clinical pathology. Direct comparison between these pathologies allowed us to find the key perturbations leading to melanoma initiation and progression. To better understand the role of aging we also integrated senescent fibroblasts into our virtual skin model.

Our primary finding is that melanoma initiation can be directly facilitated by fibroblast senescence. However, successful initiation requires the existence of preexisting mutations within the melanocyte population allowing them to exploit the stromal activation. These findings provide additional evidence demonstrating the importance of therapeutically targeting the stroma. We believe that any effective therapy will need to not only eradicate the melanoma population but also restore normal homeostatic stromal function.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 505. doi:10.1158/1538-7445.AM2011-505
Abstract 5050: Biomarkers for sensitivity to gamma secretase inhibitors in mouse models of pancreatic adenocarcinoma
Authors: Dasari Arvind , Powell Rebecca W. , Puls Lauren , Quackenbush Kevin , Tan Aik Choon , Messersmith Wells , Boylan John , Arcaroli John
Published: June 2012

Background: The Notch pathway is important in carcinogenesis and maintenance of pancreatic adenocarcinoma through its effects on cancer stem cells, cell differentiation and neoangiogenesis. RO4929097 is a novel oral inhibitor of the Notch pathway through its effects on γ-secretase, a key enzyme in this pathway. Several clinical trials are currently in progress with RO4929097 including a CTEP sponsored phase II trial in gemcitabine resistant metastatic pancreas cancer by our group. However, there are no biomarkers of sensitivity available currently for therapy with RO4929097 or other γ-secretase inhibitors (GSIs).

Methods: We intend to treat 12 human pancreas cancer cell line mouse xenografts and 6 human primary pancreas tumor explants with RO4929097 (alone or in combination with gemcitabine) to identify sensitive (defined as tumors with ≥ 50% relative tumor growth inhibition with treatment) and resistant tumors. Subsequently, we propose to identify differences in gene expression by RT-PCR (Notch pathway), gene array or gene copy number by FISH between the resistant and sensitive tumors as potential biomarkers.

Results: Of the eight pancreas cell line xenografts treated so far, four are sensitive to RO4929097. Comparison between sensitive and resistant tumors by gene array suggests that the sensitive cell lines exhibit higher levels of mesenchymal markers. In addition, two of the four human primary pancreatic explants treated thus far show intermediate sensitivity. RO4929097 in combination with gemcitabine in the four explants reveals a significant decrease in tumor growth inhibition for the combination compared to RO4929097 alone but the additive effect appears to be entirely due to gemcitabine. Similar effects were observed on tumor regrowth, upon observation after an initial period of treatment with either agents alone or in combination. In addition, future experiments to identify stem cell and/or angiogenesis specific biomarkers are also planned. Updated results will be presented at the meeting.

Acknowledgements: This work was supported by Roche Pharmaceuticals

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5050. doi:10.1158/1538-7445.AM2011-5050