Changes in intracellular Ca2+ and pH in response to thapsigargin in human glioblastoma cells and normal astrocytes

Gergely Gy Kovacs, A. Zsembery, Susan J. Anderson, Peter Komlosi, G. Yancey Gillespie, P. Darwin Bell, Dale J. Benos, Catherine M. Fuller

Research output: Contribution to journalArticle

18 Citations (Scopus)

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+ ([Ca 2+]i) and intracellular pH (pHi) regulation could be involved in tumor formation. The sarco(endo)plasmic reticulum Ca 2+-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 Ca 2+ 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 Ca 2+ homeostasis and pH regulation in tumor cells compared with normal human astrocytes.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume289
Issue number2 58-2
DOIs
Publication statusPublished - Aug 2005

Fingerprint

Thapsigargin
Glioblastoma
Astrocytes
Tumors
Cells
Reticulum
Acidification
Adenosine Triphosphatases
Neoplasms
Sodium-Hydrogen Antiporter
Calcium-Transporting ATPases
Fura-2
Amiloride
Buffers
Tumor Cell Line
Recovery
Homeostasis
Cell Line
Research

Keywords

  • BCECF
  • fura-2
  • Store-operated calcium channels

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Changes in intracellular Ca2+ and pH in response to thapsigargin in human glioblastoma cells and normal astrocytes. / Kovacs, Gergely Gy; Zsembery, A.; Anderson, Susan J.; Komlosi, Peter; Gillespie, G. Yancey; Bell, P. Darwin; Benos, Dale J.; Fuller, Catherine M.

In: American Journal of Physiology - Cell Physiology, Vol. 289, No. 2 58-2, 08.2005.

Research output: Contribution to journalArticle

Kovacs, Gergely Gy ; Zsembery, A. ; Anderson, Susan J. ; Komlosi, Peter ; Gillespie, G. Yancey ; Bell, P. Darwin ; Benos, Dale J. ; Fuller, Catherine M. / Changes in intracellular Ca2+ and pH in response to thapsigargin in human glioblastoma cells and normal astrocytes. In: American Journal of Physiology - Cell Physiology. 2005 ; Vol. 289, No. 2 58-2.
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