Anion exchanger isoform 2 operates in parallel with Na+/H+ exchanger isoform 1 during regulatory volume decrease of human cervical cancer cells

Meng Ru Shen, Robert J. Wilkins, Cheng Yang Chou, J. Clive Ellory

研究成果: Article同行評審

16 引文 斯高帕斯(Scopus)

摘要

Intracellular pH (pHi) homeostasis was investigated in human cervical cancer SiHa cells undergoing regulatory volume decrease (RVD) to determine which transport systems were involved. Using isoform-specific primers, mRNA transcripts of Na+/H+ exchanger isoform 1 (NHE1) and isoform 3 were identified by reverse transcriptase polymerase chain reaction (RT-PCR) and the results confirmed by Western immunoblotting. From anion exchanger isoforms 1-3 (AE1-3), only the mRNA transcript of AE2 was identified by RT-PCR and the identity was confirmed by digestion with a specific restriction endonuclease. SiHa cells loaded with the fluorescent dye 2′,7′-bis(carboxyethyl)-5(6)-carboxyfluorescein and resuspended in isotonic media showed a stable pHi. In contrast, a gradual internal acidification took place following resuspension in hypotonic media. The NHE inhibitors, HOE694 (10 μM) and amiloride (1 mM), showed a similar potency in enhancing the rate and extent of the hypotonicity-induced internal acidification. The absence of extracellular Na+ also substantially enhanced the acidification during RVD. These results suggest that internal acidification during RVD is mainly compensated by the operation of NHE1. Extracellular Cl- was critically necessary for the pHi acidification during RVD. The hypotonicity-induced acidification was significantly attenuated by 100 μM 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid, a concentration inhibiting more than 90% AE activity. This indicates that AE2 mediates a net Cl- influx with compensating HCO3- efflux during RVD. We conclude that AE2 operates in parallel with NHE1 to regulate pHi during RVD of human cervical cancer cells.

原文English
頁(從 - 到)52-58
頁數7
期刊FEBS Letters
512
發行號1-3
DOIs
出版狀態Published - 2002 2月 13

All Science Journal Classification (ASJC) codes

  • 生物物理學
  • 結構生物學
  • 生物化學
  • 分子生物學
  • 遺傳學
  • 細胞生物學

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