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商业专用介质对支气管上皮细胞生长和转化生长因子β1诱导的上皮-间充质转化的差异影响。

The differential effects of commercial specialized media on cell growth and transforming growth factor beta 1-induced epithelial-mesenchymal transition in bronchial epithelial cells.

机构信息

Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia.

出版信息

Mol Biol Rep. 2020 May;47(5):3511-3519. doi: 10.1007/s11033-020-05439-x. Epub 2020 Apr 11.

Abstract

Epithelial-mesenchymal transition (EMT) is one of the mechanisms that contribute to bronchial remodelling which underlie chronic inflammatory airway diseases such as chronic obstructive pulmonary disorder (COPD) and asthma. Bronchial EMT can be triggered by many factors including transforming growth factor β1 (TGFβ1). The majority of studies on TGFβ1-mediated bronchial EMT used BEGM as the culture medium. LHC-9 medium is another alternative available which is more economical but a less common option. Using normal human bronchial epithelial cells (BEAS-2B) cultured in BEGM as a reference, this study aims to validate the induction of EMT by TGFβ1 in cells cultured in LHC-9. Briefly, the cells were maintained in either LHC-9 or BEGM, and induced with TGFβ1 (5, 10 and 20 ng/ml) for 48 h. EMT induction was confirmed by morphological analysis and EMT markers expression by immunoblotting. In both media, cells induced with TGFβ1 displayed spindle-like morphology with a significantly higher radius ratio compared to non-induced cells which displayed a cobblestone morphology. Correspondingly, the expression of the epithelial marker E-cadherin was significantly lower, whereas the mesenchymal marker vimentin expression was significantly higher in induced cells, compared to non-induced cells. By contrast, a slower cell growth rate was observed in LHC-9 compared to that of BEGM. This study demonstrates that neither LHC-9 nor BEGM significantly influence TGFβ1-induced bronchial EMT. However, LHC-9 is less optimal for bronchial epithelial cell growth compared to BEGM. Thus, LHC-9 may be a more cost-effective substitute for BEGM, provided that time is not a factor.

摘要

上皮-间充质转化(EMT)是导致慢性炎症性气道疾病(如慢性阻塞性肺疾病和哮喘)支气管重塑的机制之一。支气管 EMT 可由多种因素触发,包括转化生长因子 β1(TGFβ1)。大多数关于 TGFβ1 介导的支气管 EMT 的研究都使用 BEGM 作为培养基。LHC-9 培养基是另一种更经济但不太常见的选择。本研究使用在 BEGM 中培养的正常人支气管上皮细胞(BEAS-2B)作为参考,旨在验证 TGFβ1 在 LHC-9 培养的细胞中诱导 EMT。简而言之,细胞分别在 LHC-9 或 BEGM 中维持,并分别用 TGFβ1(5、10 和 20 ng/ml)诱导 48 小时。通过形态分析和免疫印迹检测 EMT 标志物的表达来确认 EMT 诱导。在两种培养基中,与诱导前相比,用 TGFβ1 诱导的细胞呈现出梭形形态,其半径比明显更高,而诱导前的细胞呈现出鹅卵石形态。相应地,与诱导前相比,上皮标志物 E-钙黏蛋白的表达明显降低,而间充质标志物波形蛋白的表达明显升高。相比之下,与 BEGM 相比,LHC-9 中的细胞生长速度较慢。本研究表明,LHC-9 和 BEGM 均不会显著影响 TGFβ1 诱导的支气管 EMT。然而,与 BEGM 相比,LHC-9 对支气管上皮细胞生长的影响较小。因此,只要时间不是一个因素,LHC-9 可能是比 BEGM 更具成本效益的替代品。

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