Cystic fibrosis transmembrane regulator (CFTR) in human trophoblast BeWo cells and its relation to cell migration

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• 作者：G.I. Marino ; B.A. Kotsias
• 关键词：CFTR ; BeWo cells ; Ion currents ; Wound healing ; Forskolin ; CFTRinh-172
• 刊名：Placenta
• 出版年：February, 2014
• 年：2014
• 卷：35
• 期：2
• 页码：92-98
• 全文大小：951 K

文摘

Introduction

ENaC and CFTR are coexpressed in epithelia and have positive or negative functional interactions. In addition, ENaC and CFTR promote migration in placental trophoblastic cells and human airway cells, respectively. Here we tested the idea if CFTR is functionally expressed in BeWo cells, a trophoblastic cell line, and if it is involved in their migratory behavior.

Methods

CFTR expression was studied in BeWo cells with RT-PCR, biotinylation and Western blot. Ion currents were analyzed with patch clamp, and cell migration with the wound healing method.

Results

The mature CFTR 160-kDa band was present, and its localization at the surface membrane was confirmed. Forskolin (20聽渭M), an adenylate cyclase activator, was used for channel activation, and subsequently CFTR_inh-172 (2聽渭M) for its inhibition. The conductances in the presence of CFTR_inh-172 plus forskolin (16.0聽卤聽0.7 pS/pF and 32.6聽卤聽1.5 pS/pF) were significantly lower than in presence of only forskolin (29.7聽卤聽0.9 and 47.0聽卤聽2.0 pS/pF). The conductance of CFTR_inh-172 inhibited currents was 14.9聽卤聽0.7 pS/pF with a linear I-V relationship illustrating the nonrectifying properties of the CFTR. Cell migration was measured and covered 11.2聽卤聽0.4, 24.0聽卤聽1.7 and 13.9聽卤聽1.0% of the wound when cells were cultivated under control, forskolin, and forskolin plus CFTR_inh-172, respectively. Proliferation was not changed by any of the treatments.

Conclusions

Our results shows that BeWo cells functionally express the CFTR which plays a role in the wound healing increasing the cell migration process.