المؤلفون: John J. O'Leary, Dearbhaile M. O'Donnell, Sharon O'Toole, Luke A. J. O'Neill, Amanda McCann, Brian Flood, Kathryn Haley, Noreen Gleeson, Ciara Murphy, Bryan T. Hennessy, Stephen P. Finn, Stephen R. Pennington, Cathy D. Spillane, Michael F Gallagher, Britta K. Stordal, Cara Martin, Aoife A. Cooke, Kenneth J. O'Byrne, Orla Sheils, Mark Bates, Brendan Ffrench, Katie M. Costello, Niamh Kernan, Jacqui Barry-O'Crowley, Tom D'Arcy, Charles d’Adhemar, Paul Smyth

المصدر: PLoS ONE, Vol 9, Iss 6, p e100816 (2014)
PLoS ONE

مصطلحات موضوعية: Cellular differentiation, lcsh:Medicine, Autoimmunity, Carcinoma, Ovarian Epithelial, Biochemistry, Nucleic Acids, Molecular Cell Biology, Neoplasms, Glandular and Epithelial, RNA, Small Interfering, lcsh:Science, Ovarian Neoplasms, Multidisciplinary, Genomics, Middle Aged, Prognosis, Immunohistochemistry, Functional Genomics, 3. Good health, Gene Expression Regulation, Neoplastic, Phenotype, Neoplastic Stem Cells, Female, Anatomy, Stem cell, Transcriptome Analysis, Research Article, Signal Transduction, Histology, Genotype, Paclitaxel, Immunology, Biology, Cancer stem cell, Cell Line, Tumor, microRNA, Genetics, medicine, Carcinoma, Humans, Aged, lcsh:R, Biology and Life Sciences, Computational Biology, Cancer, Cell Biology, Genome Analysis, medicine.disease, Antineoplastic Agents, Phytogenic, Survival Analysis, Cystadenocarcinoma, Serous, Toll-Like Receptor 4, MicroRNAs, Drug Resistance, Neoplasm, Myeloid Differentiation Factor 88, Cancer cell, Cancer research, RNA, lcsh:Q, Genome Expression Analysis, Ovarian cancer

الوصف: The prognosis of epithelial ovarian cancer is poor in part due to the high frequency of chemoresistance. Recent evidence points to the Toll-like receptor-4 (TLR4), and particularly its adaptor protein MyD88, as one potential mediator of this resistance. This study aims to provide further evidence that MyD88 positive cancer cells are clinically significant, stem-like and reproducibly detectable for the purposes of prognostic stratification. Expression of TLR4 and MyD88 was assessed immunohistochemically in 198 paraffin-embedded ovarian tissues and in an embryonal carcinoma model of cancer stemness. In parallel, expression of TLR4 and MyD88 mRNA and regulatory microRNAs (miR-21 and miR-146a) was assessed, as well as in a series of chemosensitive and resistant cancer cells lines. Functional analysis of the pathway was assessed in chemoresistant SKOV-3 ovarian cancer cells. TLR4 and MyD88 expression can be reproducibly assessed via immunohistochemistry using a semi-quantitative scoring system. TLR4 expression was present in all ovarian epithelium (normal and neoplastic), whereas MyD88 was restricted to neoplastic cells, independent of tumour grade and associated with reduced progression-free and overall survival, in an immunohistological specific subset of serous carcinomas, p

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::054dd4c375238746aeb819e7402f12b7Test
http://europepmc.org/articles/PMC4076208?pdf=renderTest

المؤلفون: John J. O'Leary, Claudia Gasch, Michael Gallagher, Aoife A. Cooke

المصدر: Cancer Research. 72:3325-3325

مصطلحات موضوعية: Cancer Research, Toll-like receptor, Chemokine, Myeloid, Transfection, Biology, medicine.disease, medicine.anatomical_structure, Oncology, Cancer stem cell, Cell culture, Cancer cell, Immunology, medicine, Cancer research, biology.protein, Ovarian cancer

الوصف: Ovarian cancer is the leading cause of gynaecological cancer death worldwide. This high mortality is caused in large part by development of chemoresistant recurrent disease. In primary tumours differentiation status of the tumour is considered a key prognostic factor however in recurrent disease this is no longer the case, suggesting that in recurrent tumours the differentiation status of the tumour has no bearing on the response to chemotherapy. The origins of recurrent disease may be explained by the cancer stem cell (CSC) theory. CSCs are a minority population of cancer cells with stem like properties including enhanced proliferation. The adaptor molecule myeloid differentiation-primary response gene (88) (MyD88), is a key constituent of the toll like receptor 4 (TLR4) pathway. In normal circumstances TLR pathways mediate the body's response to pathogens; however MyD88 has been recently suggested as a possible marker of cancer stemness in ovarian cancer. This work investigates the involvement of MyD88 in the CSC response to chemotherapy, differentiation stimulus and hypoxia treatment. Cell line experiments were carried out in two CSC lines: NTera2 and 2102Ep. NTera2 cells readily differentiate in response to retinoic acid (RA) treatment while 2102Ep cells are nullipotent and resist differentiation via RA. NTera2 cells are more chemosensitive than 2102Ep cells. Cells were incubated in the presence of both retinoic acid and a platinum based chemotherapy drug, both in isolation and in various combinations. RNA was isolated from treated cells, cDNA was synthesised and interrogated for TLR4/MyD88 expression via Q-PCR. To assess whether MyD88 was necessary and/or sufficient for the normal cell response, MyD88 was knocked down in cells via siRNA transfection, and overexpressed via insertion of an overexpression plasmid. The effect of these alterations on the cell lines response to chemotherapy, differentiation stimulus and hypoxia was then measured. The downstream effect of ablation of the MyD88 signal was also assessed using an Affymetrix array, and the chemokine and cytokine expression profile of both cell lines in all treatments was assessed using Quantibody arrays from RayBiotech. MyD88 expression was shown to be affected by chemotherapy, differentiation stimulus and hypoxic treatments. Furthermore when cells were pre-treated with differentiation stimulus or chemotherapy, their normal responses were altered. So far, our work has shown that ablation of the MyD88 signal in nullipotent 2102Ep cells removes their ability to resist differentiation via RA. To close this mechanism we have also shown that overexpression of MyD88 in pluripotent NTera2 cells allows them to resist differentiation via RA. Functional analysis of the role of MyD88 in chemotherapy and hypoxia resistance is currently being assessed. This and the mechanisms involved will be detailed, along with the chemokine/cytokine profile of treated 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 3325. doi:1538-7445.AM2012-3325

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::d73aaeb671e041dc93763428c9a200d6Test
https://doi.org/10.1158/1538-7445.am2012-3325Test

المؤلفون: Sharon O'Toole, Michael Gallagher, John J. O'Leary, Aoife A. Cooke, Britta K. Stordal, Orla Shiels, Brendan Ffrench

المصدر: Cancer Research. 72:3382-3382

مصطلحات موضوعية: Oncology, Cancer Research, medicine.medical_specialty, business.industry, Cancer stem cell, Internal medicine, Medicine, business, Ovarian cancer, medicine.disease

الوصف: Background: Tumorgenicity studies show that Cancer Stem Cells (CSCs) but not their differentiated counterparts efficiently generate tumors. Therapeutically targeting CSCs could remove the malignant potential from tumours, while circumventing chemoresistance, relapse and metastasis. CSCs are difficult to study as they usually represent a small fraction of tumors. We will present a systematic approach we developed to investigate CSCs within any malignancy. We will present the data obtained thus far in the application of this system to the study of Ovarian CSCs. Methods: We developed a four phase model for investigating CSCs: Screening - Validation - Characterisation - Identification of novel therapeutic targets. Six cell lines modelling various stages of ovarian cancer (A2780, A2780cis, IGROV-1, IGROV(CDDP), SK-OV-3 and 59M) and one cell line (HIO-80) modelling non malignant ovarian surface epithelium were screened for the presence of CSCs and somatic stem cells respectively. Three flow cytometry based screens were implemented; ALDEFLUOR, Hoechst Side Population and Cell Surface Protein Screens. Cells of interest were isolated via Fluorescence Activated Cell Sorting. Mouse Tumorgenicity and Asymmetric Division Assays have been developed to validate putative CSCs (pCSCs) as true CSCs. Hypoxic growth, chemotherapeutic and matrigel invasion assays have all been developed to investigate the role of Ovarian CSCs in these commonly observed phenotypes of Ovarian cancer. Results: Six pCSC populations have been identified. No overlap has been observed between the three screening approaches. ALDEFLUOR identified pCSCs within A2780 and A2780cis. Hoechst Side Population identified pCSCs within IGROV-1 and IGROV(CDDP). CD44/CD117 identified pCSCs within SK-OV-3 and 59M. No somatic stem cells were identified within HIO-80. To date pCSC and non-pCSCs have been sorted from two cell lines. In both models the isolated pCSCs regenerated the non-pCSC phenotype. Both the isolated pCSCs and non-pCScs from each model are resistance to cisplatin and paclitaxel. In both cases the parent lines were chemoresistant, so this finding is not unexpected. All pCSCs are currently being validated via Mouse Tumourgenicity and Single Cell Asymmetric Division Assays. All validated CSCs will be described. Conclusion: A systematic approach has been established for the isolation and study of CSCs. Each ‘type’ of ovarian CSC appears to be mutually exclusive. This may reflect different stages/histologies of ovarian disease, or perhaps the selective conditions of tissue culture. pCSCs have been identified within chemosensitive cell lines. Investigation of the differences between chemoresistant and chemosensitive CSCs could elucidate the mechanisms behind chemoresistant relapse. 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 3382. doi:1538-7445.AM2012-3382

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::6a68d90aba969bb8a28e93178442b62bTest
https://doi.org/10.1158/1538-7445.am2012-3382Test