Establishment of Immortalized Laryngeal Epithelial Cells Transfected with Bmi1
| Autor: | Juan Lu, Jia-Jie Tan, Lu Wang, Huai-Hong Chen, Xiang-Ping Li, Xiong Liu, Wendong Tian, Yuan-Feng Dai, Ting-Ting Mo |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Male
Cell Blotting Western Biomedical Engineering lcsh:Medicine Mice Nude Stratified squamous epithelium macromolecular substances Biology Flow cytometry 03 medical and health sciences Mice 0302 clinical medicine Proto-Oncogene Proteins medicine Animals Humans Telomerase reverse transcriptase immortal 030223 otorhinolaryngology Cellular Senescence Cell Proliferation Polycomb Repressive Complex 1 Transplantation Mice Inbred BALB C laryngeal epithelial cells medicine.diagnostic_test Cell growth lcsh:R Cell Cycle Epithelial Cells Cell Biology Transfection Middle Aged Bmi1 medicine.anatomical_structure Cell culture Laryngeal Mucosa 030220 oncology & carcinogenesis Cancer research Original Article Female Immortalised cell line |
| Zdroj: | Cell Transplantation Cell Transplantation, Vol 29 (2020) |
| ISSN: | 1555-3892 0963-6897 |
| Popis: | Primary laryngeal epithelial cells are essential to exploring the mechanisms of laryngeal and voice disorders; however, they are difficult to study and apply because of their limited life span. The purpose of this study was to develop a stable and reliable in vitro model for the comprehensive study of the pathogenesis of laryngeal and voice diseases. The pLVTHM-Bmi1 plasmid was constructed and used to immortalize primary laryngeal epithelial cells by lentiviral infection. The expressions of Bmi1, human telomerase reverse transcriptase (hTERT), p53, and pRB pathway proteins were detected by western blotting. Functional characteristics of the immortalized cell lines were verified by cell senescence β-galactosidase staining, 5-ethynyl-2′-deoxyuridine cell proliferation test, and flow cytometry. We successfully introduced Bmi into human subglottic (hSG) cells and human ventricle (hV) cells. Both the human immortalized subglottic Bmi1 (hSG-Bmi1) cell line and the human immortalized ventricle Bmi1 (hV-Bmi1) cell line maintained normal epithelial morphology and divided successfully after more than 20 culture passages. As Bmi1 was overexpressed in these cells, the expression of human telomerase reverse transcriptase (hTERT) and phosphorylated Rb increased while p16 and p21 decreased. Following Bmi1-mediated immortalization, cell senescence decreased significantly, and cell proliferation was accelerated. Tumor formation was not observed for hSG, hV, or hSG-Bmi1, and hV-Bmi1 cells in nude mice. hSG-Bmi1 cells dominated by stratified squamous epithelium and hV-Bmi1 cells dominated by columnar cells were established. The new cell lines lay a foundation for the study of the pathogenic mechanisms of laryngeal and voice diseases. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|