Characterization of cancer stem cells from non-small cell lung cancer
[EN] Background: Despite the advances in the molecular characterization of lung cancer, chemoresistance, tumor progression and metastasis make of lung cancer the first cause of death cancer-related worldwide. Cancer stem cells (CSCs) are small subpopulations of stem-like cells with self-renewal, dif...
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| Tipo de recurso: | tesis de maestría |
| Fecha de publicación: | 2016 |
| País: | España |
| Institución: | Universitat Politècnica de València (UPV) |
| Repositorio: | RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia |
| Idioma: | inglés |
| OAI Identifier: | oai:riunet.upv.es:10251/146816 |
| Acceso en línea: | https://riunet.upv.es/handle/10251/146816 |
| Access Level: | acceso abierto |
| Palabra clave: | Célula madre tumoral Cáncer de pulmón no microcítico Expresión Génica Vía Notch Vía Wnt Vía Hedgehog Cancer Stem Cells Non-Small Cell Lung Cancer Relative Gene Expression Notch Pathway Wnt Pathway Hedgehog Pathway. BIOLOGIA CELULAR Máster Universitario en Biotecnología Biomédica-Màster Universitari en Biotecnologia Biomèdica |
| Sumario: | [EN] Background: Despite the advances in the molecular characterization of lung cancer, chemoresistance, tumor progression and metastasis make of lung cancer the first cause of death cancer-related worldwide. Cancer stem cells (CSCs) are small subpopulations of stem-like cells with self-renewal, differentiation and tumorigenic properties that constitute a promising target, but remain largely unknown. The aim of this study was to isolate and analyze gene expression of CSCs from lung cancer cell-lines and tumor-tissue from resectable NSCLC patients. Methods: This study was performed on cells from NSCLC tumor samples and cell lines (H1650, H1993, A549 and PC9) grown in monolayer and as spheroids. The expression of: CSC-markers (CD133, EPCAM1, ALDH1A1, CD166, ABCG2, CD44, MUC1, BMI1); pluripotency (KLF4, OCT4, NANOG, SOX2, MYC, CCND1); cell cycle (CDKN1A, CDKN2A, MDM2, WEE1); invasiveness (CDH1, VIM, SNAI1, MMP2, MMP9, CEACAM5); Notch pathway (NOTCH1, NOTCH2, NOTCH3, DLL1, DLL4, HEY1, HES1); Wnt pathway (WNT1, WNT2, WNT3, WNT5A, CTNBB1, DKK1, FZD7) and Hedgehog pathway (SMO, PTCH1, SHH, GLI1) were analyzed by quantitative real-time PCR (qPCR). Housekeeping genes ACTB, CDKN1B and GUSB were used as endogenous controls for relative expression calculation. Results: Lung tumorspheres had increased expression of EPCAM1, CD44, ALDH1A1 and CDKN1A (p= 0.028, p= 0.021, p= 0.043 and p= 0.021, respectively) when compared to their paired-adherent cells. In addition, epithelial to mesenquimal transition (EMT) inducer SNAI1 was overexpressed (p= 0.011) in tumorspheres. Regarding Notch pathway, DLL4, NOTCH1 and NOTCH2 showed higher expression in spheroids (p= 0.028, p= 0.038 and p= 0.036, respectively). In Wnt pathway, we found higher expression levels of WNT3, CTNBB1 and GSK3B (p= 0.021, p= 0.008 and p= 0.021, respectively) in lungspheres, whereas the activator of the non-canonical Wnt pathway, WNT5A, tended to be less expressed in spheroids compared to adherent-cultured cells. No significant differences were found in other analyzed genes. Conclusions: Lung spheroids from cancer cell lines and primary tumors showed increased levels of CSC-markers. Genes related to Notch and Wnt were found to be more expressed in tumorspheres, suggesting these pathways as interesting lung-CSC targets. |
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