Expression of polycomb protein BMI-1 maintains the plasticity of basal bronchial epithelial cells.
Expression of polycomb protein BMI-1 maintains the plasticity of basal bronchial epithelial cells.
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DOI：
10.14814/phy2.12847
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发表时间：
2016-08
影响因子：
2.5
通讯作者：
Sayers I
中科院分区：
文献类型：
--
作者：
Torr E;Heath M;Mee M;Shaw D;Sharp TV;Sayers I
呼吸道上皮细胞在呼吸道疾病中发生改变，被认为是疾病病因的一部分。疾病研究的一个警告是，从患者身上分离支气管上皮细胞的技术是侵入性的，细胞的寿命有限。本研究的目的是广泛表征原代人支气管上皮细胞的可塑性，这些细胞已被工程化以延迟细胞衰老，包括这些细胞的分化能力。使用病毒载体系统将细胞工程化以表达BMI-1或hTERT。在传代（p）早期（p5）、中期（p10）和晚期（p15）阶段表征细胞：BMI-1、p16和CK 14蛋白表达、活力和在气-液界面（ALI）分化的能力，使用一系列技术，包括免疫组织化学（IHC）、免疫荧光（IF）、跨上皮电阻（TEER）、扫描电子显微镜（SEM），MUC 5AC和β微管蛋白（BTUB）染色。表达BMI-1的细胞保持BMI-1蛋白和上皮标志物CK 14的升高水平，并显示出对p16的抑制。BMI-1表达细胞具有活力优势，在ALI时分化，并具有正常的核型。相比之下，hTERT表达细胞的活力降低，分化有限，核型异常。因此，我们提供了在ALI模型背景下表达BMI-1的细胞的可塑性的广泛表征。这些细胞保留了野生型细胞的特性，可能有助于在体外持续表征呼吸系统疾病机制。
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.