Analysis of the differential secretome of nasopharyngeal carcinoma cell lines CNE-2R and CNE-2
| Autor: | Hao Chen, Ling Li, Qi-Yan Mo, Song Qu, Ya Guo, Ze-Tan Chen, Xiao-Dong Zhu, Jun Yin, Fang Su, Wei Zhao |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Fetal Proteins
Cancer Research Proteome Cell Survival Cell Adhesion Molecules Neuronal Quantitative proteomics Nasopharyngeal neoplasm Biology Radiation Tolerance Antigens CD Cancer stem cell Cell Line Tumor Radioresistance Biomarkers Tumor otorhinolaryngologic diseases medicine Humans Exosomal secretion Nasopharyngeal Carcinoma Carcinoma Nasopharyngeal Neoplasms General Medicine medicine.disease Molecular biology stomatognathic diseases Secretory protein Oncology Nasopharyngeal carcinoma Neoplastic Stem Cells |
| Zdroj: | Oncology Reports. 34:2477-2488 |
| ISSN: | 1791-2431 1021-335X |
| Popis: | Radioresistance is the major cause of poor prognosis in nasopharyngeal carcinoma (NPC). To identify and characterize the secretome associated with NPC radioresistance, we compared the conditioned serum-free medium of radioresistant CNE-2R cells with that of the parental radiosensitive CNE-2 cells using isobaric tags for relative and absolute quantitation (iTRAQ) with liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) quantitative proteomics. Before proceeding to quantitative proteomics, we investigated the survival curves of CNE-2R and CNE-2 cells by colony formation assay, and the CNE-2R survival curves were significantly higher than those for CNE-2. In total, 3,581 proteins were identified in the quantitative proteomics experiments, and 40 proteins exhibited significant differences between the CNE-2R and CNE-2 cells. Twenty-six of the 40 proteins were secreted by classical, non-classical, or exosomal secretion pathways. To verify the reliability of iTRAQ quantitative proteomics, we applied western blotting (WB) to study the secretory protein expression of fibrillin-2, CD166, sulfhydryl oxidase 1 and cofilin-2, which are involved in cell adhesion, migration and invasion. The WB results showed that fibrillin-2 (p=0.017) and sulfhydryl oxidase 1 (p=0.000) were highly expressed in the CNE-2 cells, while CD166 (p=0.012) and cofilin-2 (p=0.003) were highly expressed in the CNE-2R cells, which was in accordance with iTRAQ quantitative proteomics. Finally, a phenotypic subset of CD166-positive NPC cells was verified by immunocytochemistry. In summary, we defined a collection of secretory proteins that may be relevant to the radioresistance in NPC cells, and we determined that CD166, which is widely used as a positive marker of cancer stem cells, is expressed in NPC cells. |
| Databáze: | OpenAIRE |
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