Expressional and Functional Characterization of Intracellular pH Regulators and Effects of Ethanol in Human Oral Epidermoid Carcinoma Cells
Autor: | Yi-Ting Tsai, Shu-Fu Huang, Shih-Hurng Loh, Shiao-Pieng Lee, Yen-Lin Chen, Shih-Chi Chao |
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Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Physiology Intracellular pH Human Oral Epidermoid Carcinoma (OEC-M1) Cells lcsh:Physiology lcsh:Biochemistry 03 medical and health sciences chemistry.chemical_compound Western blot Cell Line Tumor medicine Humans lcsh:QD415-436 Na+-H+ Exchanger (NHE) Ethanol Dose-Response Relationship Drug medicine.diagnostic_test lcsh:QP1-981 Transporter Hydrogen-Ion Concentration Intracellular pH Transporters Molecular biology Neoplasm Proteins Gene Expression Regulation Neoplastic Dose–response relationship BCECF 030104 developmental biology chemistry Epidermoid carcinoma Cell culture DIDS Na+-HCO3- Cotransporter (NBC) Carcinoma Squamous Cell Mouth Neoplasms |
Zdroj: | Cellular Physiology and Biochemistry, Vol 47, Iss 5, Pp 2056-2068 (2018) |
ISSN: | 1421-9778 1015-8987 |
Popis: | Background/Aims: To functionally characterize intracellular pH (pHi) regulating mechanisms, such as Na+-H+ exchanger (NHE) and Na+-HCO3- co-transporter (NBC), and further examine effects of ethanol on the pHi regulating mechanism in human oral epidermoid carcinoma (OEC-M1) cells. Methods: OEC-M1 cells were a gift from Tri-Service General Hospital. Changes of pHi were detected by microspectrofluroimetry with a pH-sensitive fluorescent dye, BCECF. Isoforms of transporters were examined by Western blot technique. Results: i) the steady-state pHi value shifted from alkaline (7.35∼7.49) to acidic (7.0∼7.03) following acid/base impacts; ii) in HEPES-buffer system, pHi recovery following induced-acidification was totally blocked by either removing [Na]o+ or adding HOE 694 (a NHE1 specific inhibitor), which demonstrates existence of NHE1; iii) in HCO3-/CO2-buffer system, the pHi recovery following induced-acidification was entirely blocked by either removing [Na]o+ or adding HOE 694 plus DIDS (a NBC specific inhibitor), which suggests existence of Na+- and HCO3–dependent acid-extruder, i.e. NBC; iv) the isoforms of the two acid extruders were NHE1, NBCn1, NBCe1 and NDCBE; v) ethanol (10-1000 mM) showed a biphasic and concentration-dependent effect on resting pHi (i.e. increase then decrease) by changing the activity of NHE1 and NBC accordingly; vi) treatment with ethanol for 24 hr (> 300 mM) significantly inhibited the expression of NHE1, NBCn1 and NDCBE, while up-regulated NBCe1. Conclusions: Ethanol affects pHi in a concentration-dependent manner by changing function and expression of NHE1 and NBC isoforms in OEC-M1 cells. |
Databáze: | OpenAIRE |
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