Changes in intracellular Ca2+ and pH in response to thapsigargin in human glioblastoma cells and normal astrocytes |
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Authors: | Kovacs Gergely Gy Zsembery Akos Anderson Susan J Komlosi Peter Gillespie G Yancey Bell P Darwin Benos Dale J Fuller Catherine M |
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Institution: | Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL 35294, USA. |
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Abstract: | Despite extensive work in the field of glioblastoma research no significant increase in survival rates for this devastating disease has been achieved. It is known that disturbance of intracellular Ca2+ (Ca2+]i) and intracellular pH (pHi) regulation could be involved in tumor formation. The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) is a major regulator of Ca2+]i. We have investigated the effect of inhibition of SERCA by thapsigargin (TG) on Ca2+]i and pHi in human primary glioblastoma multiforme (GBM) cells and GBM cell lines, compared with normal human astrocytes, using the fluorescent indicators fura-2 and BCECF, respectively. Basal Ca2+]i was higher in SK-MG-1 and U87 MG but not in human primary GBM cells compared with normal astrocytes. However, in tumor cells, TG evoked a much larger and faster Ca2+]i increase than in normal astrocytes. This increase was prevented in nominally Ca2+-free buffer and by 2-APB, an inhibitor of store-operated Ca2+ channels. In addition, TG-activated Ca2+ influx, which was sensitive to 2-APB, was higher in all tumor cell lines and primary GBM cells compared with normal astrocytes. The pHi was also elevated in tumor cells compared with normal astrocytes. TG caused acidification of both normal and all GBM cells, but in the tumor cells, this acidification was followed by an amiloride- and 5-(N,N-hexamethylene)-amiloride-sensitive recovery, indicating involvement of a Na+/H+ exchanger. In summary, inhibition of SERCA function revealed a significant divergence in intracellular Ca2+ homeostasis and pH regulation in tumor cells compared with normal human astrocytes. fura-2; BCECF; store-operated calcium channels |
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