Bischof G, Wenzl E, Weinlich M, Hamilton G, Feil W, Schiessel R
1. Chirurgische Universitätsklinik, Wien.
Wien Klin Wochenschr. 1990 Jun 22;102(13):369-75.
Intracellular pH (pHi) regulation is essential for basic functioning of the cell and activation of pHi regulatory mechanisms appears to be involved in the initial stage of cell division. Little is known about pHi regulation in human colonic carcinoma cells. We investigated SW-620 (CCL 227) cells, a cell-line derived from a human colonic adenocarcinoma. pHi changes were recorded by computer-assisted spectrofluorimetric monitoring of the pH-sensitive, fluorescent dye BCECF (2',7'-bis(carboxyethyl)- 5(6)carboxyfluorescein). Resting pHi in HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffered solution was 7.53 +/- 0.01. Intracellular acidification after an ammonium prepulse produced a pHi decline of 0.5 units and pHi returned to normal value in NaCl Ringer's. Both 1 mM amiloride and Na-free solution completely inhibited recovery for 8 minutes. This inhibition was reversible in NaCl Ringer's. Na-free solution led to a pHi decrease to 7.39 +/- 0.04 after 16 min, pHi was also lowered by 8 minute incubation of cells with 1 mM amiloride (7.40 +/- 0.02). In HCO3/CO2-buffered solution resting pHi was 7.42 +/- 0.01 (n = 35). Recovery from an acute acid load, induced by NH4 prepulse or switching from HEPES- to bicarbonate-buffered solution, was Na dependent, Cl independent, reversible and only partially blocked by 1 mM amiloride - pHi slowly recovered from 6.83 +/- 0.03 to 7.00 +/- 0.06 in 8 minutes. In the presence of amiloride and 200 microns H2DIDS (dihydro-4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid) pHi recovery was completely inhibited for 8 minutes. In Na-free solution pHi decreased from 7.44 +/- 0.04 to 7.29 +/- 0.03 within 8 minutes.(ABSTRACT TRUNCATED AT 250 WORDS)
细胞内pH值（pHi）调节对于细胞的基本功能至关重要，pHi调节机制的激活似乎参与细胞分裂的初始阶段。关于人结肠癌细胞中的pHi调节知之甚少。我们研究了SW - 620（CCL 227）细胞，这是一种源自人结肠腺癌的细胞系。通过计算机辅助的荧光分光光度法监测pH敏感荧光染料BCECF（2'，7'-双（羧乙基）-5（6）羧基荧光素）来记录pHi变化。在HEPES（N - 2 - 羟乙基哌嗪 - N'-2 - 乙磺酸钠）缓冲溶液中的静息pHi为7.53±0.01。铵预脉冲后的细胞内酸化使pHi下降0.5个单位，在NaCl林格氏液中pHi恢复到正常值。1 mM氨氯吡脒和无钠溶液在8分钟内完全抑制了恢复。这种抑制在NaCl林格氏液中是可逆的。无钠溶液在16分钟后使pHi降至7.39±0.04，用1 mM氨氯吡脒孵育细胞8分钟也使pHi降低（7.40±0.02）。在HCO3/CO2缓冲溶液中静息pHi为7.42±0.01（n = 35）。由NH4预脉冲或从HEPES缓冲溶液切换到碳酸氢盐缓冲溶液引起的急性酸负荷后的恢复是钠依赖性的、氯非依赖性的、可逆的，并且仅部分被1 mM氨氯吡脒阻断 - pHi在8分钟内从6.83±0.03缓慢恢复到7.00±0.06。在氨氯吡脒和200微摩尔H2DIDS（二氢 - 4,4'-二异硫氰酸根合芪 - 2,2'-二磺酸）存在下，pHi恢复被完全抑制8分钟。在无钠溶液中，pHi在8分钟内从7.44±0.04降至7.29±0.03。（摘要截断于250字）
