Changes in intracellular Ca2+ and pH in response to thapsigargin in human glioblastoma cells and normal astrocytes.
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 Ca(2+) ([Ca(2+)](i)) and intracellular pH (pH(i)) regulation could be involved in tumor formation. The sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) is a major regulator of [Ca(2+)](i). We have investigated the effect of inhibition of SERCA by thapsigargin (TG) on [Ca(2+)](i) and pH(i) 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 [Ca(2+)](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 [Ca(2+)](i) increase than in normal astrocytes. This increase was prevented in nominally Ca(2+)-free buffer and by 2-APB, an inhibitor of store-operated Ca(2+) channels. In addition, TG-activated Ca(2+) influx, which was sensitive to 2-APB, was higher in all tumor cell lines and primary GBM cells compared with normal astrocytes. The pH(i) 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 Ca(2+) homeostasis and pH regulation in tumor cells compared with normal human astrocytes.
Kovacs
Gergely Gy
GG
Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Zsembery
Akos
A
Anderson
Susan J
SJ
Komlosi
Peter
P
Gillespie
G Yancey
GY
Bell
P Darwin
PD
Benos
Dale J
DJ
Fuller
Catherine M
CM
D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007424:Intracellular Fluid / Q000737:chemistry; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D019284:Thapsigargin / Q000494:pharmacology*
D015220:Calcium Channels; D004791:Enzyme Inhibitors; D019284:Thapsigargin; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases; D002118:Calcium