Expression of polycomb protein BMI‐1 maintains the plasticity of basal bronchial epithelial cells

Autor: Elizabeth E. Torr, Maureen Mee, Ian Sayers, Tyson V. Sharp, Dominick E. Shaw, Meg Heath
Jazyk: angličtina
Rok vydání: 2016
Předmět:
0301 basic medicine
Adult
Male
Telomerase
Pathology
medicine.medical_specialty
Physiology
Cellular differentiation
Cell
Immunology
Cell Plasticity
Karyotype
Bronchi
Ageing and Degeneration
Biology
In Vitro Techniques
03 medical and health sciences
0302 clinical medicine
Physiology (medical)
medicine
Humans
Telomerase reverse transcriptase
bronchial epithelial cells
Cellular Senescence
Original Research
Polycomb Repressive Complex 1
Respiratory Conditions Disorder and Diseases
Genes
p16
Lentivirus
Cell Differentiation
Epithelial Cells
Middle Aged
In vitro
BMI‐1
Peptide Fragments
Cell biology
030104 developmental biology
medicine.anatomical_structure
Retroviridae
030220 oncology & carcinogenesis
Amniotic epithelial cells
Regulatory Pathways
plasticity
Respiratory epithelium
Genetic Engineering
lifespan
Zdroj: Physiological Reports
ISSN: 2051-817X
Popis: The airway epithelium is altered in respiratory disease and is thought to contribute to disease etiology. A caveat to disease research is that the technique of isolation of bronchial epithelial cells from patients is invasive and cells have a limited lifespan. The aim of this study was to extensively characterize the plasticity of primary human bronchial epithelial cells that have been engineered to delay cell senescence including the ability of these cells to differentiate. Cells were engineered to express BMI‐1 or hTERT using viral vector systems. Cells were characterized at passage (p) early (p5), mid (p10), and late (p15) stage for: BMI‐1, p16, and CK14 protein expression, viability and the ability to differentiate at air–liquid interface (ALI), using a range of techniques including immunohistochemistry (IHC), immunofluorescence (IF), transepithelial electrical resistance (TEER), scanning electron microscopy (SEM), MUC5AC and beta tubulin (BTUB) staining. BMI‐1‐expressing cells maintained elevated levels of the BMI‐1 protein and the epithelial marker CK14 and showed a suppression of p16. BMI‐1‐expressing cells had a viability advantage, differentiated at ALI, and had a normal karyotype. In contrast, hTERT‐expressing cells had a reduced viability, showed limited differentiation, and had an abnormal karyotype. We therefore provide extensive characterization of the plasticity of BMI‐1 expressing cells in the context of the ALI model. These cells retain properties of wild‐type cells and may be useful to characterize respiratory disease mechanisms in vitro over sustained periods.
Databáze: OpenAIRE
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