NHE1, NHE2, and NHE4 contribute to regulation of cell pH in T84 colon cancer cells

被引:35
作者
Beltran, Ana Rosa [1 ]
Ramirez, Marco Antonio [1 ,2 ]
Carraro-Lacroix, Luciene R. [1 ]
Hiraki, Yumi [1 ]
Reboucas, Nancy Amaral [1 ]
Malnic, Gerhard [1 ]
机构
[1] Univ Sao Paulo, Inst Ciencias Biomed, Dept Fisiol & Biofis, BR-05508900 Sao Paulo, Brazil
[2] Univ Antofagasta, Fac Hlth Sci, Biomed Dept, Antofagasta, Chile
来源
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY | 2008年 / 455卷 / 05期
基金
巴西圣保罗研究基金会;
关键词
T84 colon cells; NHE isoforms; NHE4; cell volume; cell pH;
D O I
10.1007/s00424-007-0333-0
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
The isoforms of the Na+/H+ exchanger present in T84 human colon cells were identified by functional and molecular methods. Cell pH was measured by fluorescence microscopy using the probe BCECF. Based on the pH recovery after an ammonium pulse and determination of buffering capacity of these cells, the rate of H+ extrusion (J(H)) was 3.68 mM/min. After the use of the amiloride derivative HOE-694 at 25 mu M, which inhibits the isoforms NHE1 and NHE2, there remained 43% of the above transport rate, the nature of which was investigated. Evidence of the presence of NHE1, NHE2, and NHE4 was obtained by reverse transcriptase polymerase chain reaction (RT-PCR) (mRNA) and Western blot. There was no decrease of J(H) by the NHE3 inhibitor S3226 (1 mu M) and no evidence of this isoform by RT-PCR was found. The following functional evidence for the presence of NHE4 was obtained: 25 mu M EIPA abolished J(H) entirely, but NHE4 was not inhibited at 10 mu M; substitution of Na by K increased the remainder, a property of NHE4; hypertonicity also increased this fraction of J(H). Cl--dependent NHE was not detected: in 0 Cl- solutions J(H) was increased and not reduced. In 0 Cl- cell volume decreased significantly, which was abolished by the Cl- channel blocker NPPB, indicating that the 0 Cl- effect was because of reduction of cell volume. In conclusion, T84 human colon cells contain three isoforms of the Na+/H+ exchanger, NHE1, NHE2, and NHE4, but not the Cl-dependent NHE.
引用
收藏
页码:799 / 810
页数:12
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