RRC ID 37600
Author Takano M, Sugimoto N, Ehrhardt C, Yumoto R.
Title Functional Expression of PEPT2 in the Human Distal Lung Epithelial Cell Line NCl-H441.
Journal Pharm Res
Abstract PURPOSE:The peptide transporter PEPT2 is expressed in alveolar type II epithelial cells. So far, however, no appropriate alveolar epithelial cell line for studying PEPT2 function has been known. In this study, we examined the functional expression of PEPT2 in the human distal lung epithelial cell line NCl-H441 (H441).
METHODS:Expression of PEPT2 mRNA and protein was examined in H441 cells. Transport function of PEPT2 was studied using glycylsarcosine (Gly-Sar) as a substrate.
RESULTS:Lamellar bodies were well developed in H441 cells and mRNA expression of type II cell markers and PEPT2 increased during time in culture. PEPT2 protein expression was confirmed in H441 cells, but not in A549 cells, by immunostaining and Western blotting. The uptake of Gly-Sar in H441 cells was inhibited by cefadroxil, and the cefadroxil-sensitive uptake was pH-dependent and peaked at pH 6.5. Gly-Sar uptake in H441 cells showed saturation kinetics with a Km value of 112.5 μM. In addition, apical-to-basal, but not basal-to-apical, transport of cephalexin across H441 cell monolayers was sensitive to cefadroxil.
CONCLUSIONS:PEPT2 is functionally expressed in H441 cells, making the cell line a good in vitro model to study PEPT2 function and its regulation in human distal lung.
Volume 32(12)
Pages 3916-26
Published 2015-12-1
DOI 10.1007/s11095-015-1751-x
PII 10.1007/s11095-015-1751-x
PMID 26168863
MeSH Cell Line Dipeptides / metabolism* Epithelial Cells / cytology Epithelial Cells / metabolism* Gene Expression Humans Lung / cytology* Lung / metabolism RNA, Messenger / genetics Symporters / analysis Symporters / genetics* Symporters / metabolism*
IF 3.242
Times Cited 13
Human and Animal Cells A549