Abstract
Background and purpose: Cisplatin drives specific types of tumour cells to apoptosis. In this study we investigate the involvement of intracellular calcium ([Ca 2+] i) in triggering apoptosis in two different cell lines. As cisplatin is used for the treatment of several forms of cancer we choose HeLa-S3 and U2-OS as two examples of tumour cell lines. Experimental approach: Cisplatin (1nM-10μM) was applied to HeLa-S3 and U2-OS cells and [Ca 2+] i measured with fluo-4, using laser scanning microscopy. Inositol-1,4,5-trisphosphate (IP 3) receptors were visualized with immunostaining. Membrane conductances were measured with patch-clamp techniques. Levels of calpain and caspases were assessed by western blots and apoptotic cells were stained with Hoechst 33342 and counted. Key results: Cisplatin increases [Ca 2+] i concentration- dependently in HeLa-S3 but not in U2-OS cells. This elevation of [Ca 2+] i depended on extracellular Ca 2+ but was reduced by the IP 3 receptor blocker, 2-APB. This effect was not due to a Ca 2+ release triggered by Ca 2+ entry. Immunostaining showed IP 3-receptors (type1-3) at the cellular membrane of HeLa-S3 cells, but not in U2-OS cells. Electrophysiological experiments showed an increased membrane conductance with cisplatin only when Ca 2+ was present extracellularly. Increase of [Ca 2+] i was related to the activation of calpain but not caspase-8 and triggered apoptosis in HeLa-S3 but not in U2-OS cells. Conclusions and implications: Our observations on the activation of IP 3-receptors, calcium entry and apoptotic rate by cisplatin in specific carcinogenic cells might open new possibilities in the treatment of some forms of cancer.
| Original language | English |
|---|---|
| Pages (from-to) | 1176-1186 |
| Number of pages | 11 |
| Journal | British Journal of Pharmacology |
| Volume | 151 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Aug 2007 |
| Externally published | Yes |
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Keywords
- [Ca ]
- Anticancer drugs
- Apoptosis
- Calcium signalling
- Calcium stores
- Calpain
- Carboplatin
- Cisplatin
- HeLa-S3
- IP -receptor
- U2-OS
ASJC Scopus subject areas
- Pharmacology
Cite this
IP 3 receptor antagonist, 2-APB, attenuates cisplatin induced Ca 2+-influx in HeLa-S3 cells and prevents activation of calpain and induction of apoptosis. / Splettstoesser, F.; Florea, A. M.; Busselberg, Dietrich.
In: British Journal of Pharmacology, Vol. 151, No. 8, 08.2007, p. 1176-1186.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - IP 3 receptor antagonist, 2-APB, attenuates cisplatin induced Ca 2+-influx in HeLa-S3 cells and prevents activation of calpain and induction of apoptosis
AU - Splettstoesser, F.
AU - Florea, A. M.
AU - Busselberg, Dietrich
PY - 2007/8
Y1 - 2007/8
N2 - Background and purpose: Cisplatin drives specific types of tumour cells to apoptosis. In this study we investigate the involvement of intracellular calcium ([Ca 2+] i) in triggering apoptosis in two different cell lines. As cisplatin is used for the treatment of several forms of cancer we choose HeLa-S3 and U2-OS as two examples of tumour cell lines. Experimental approach: Cisplatin (1nM-10μM) was applied to HeLa-S3 and U2-OS cells and [Ca 2+] i measured with fluo-4, using laser scanning microscopy. Inositol-1,4,5-trisphosphate (IP 3) receptors were visualized with immunostaining. Membrane conductances were measured with patch-clamp techniques. Levels of calpain and caspases were assessed by western blots and apoptotic cells were stained with Hoechst 33342 and counted. Key results: Cisplatin increases [Ca 2+] i concentration- dependently in HeLa-S3 but not in U2-OS cells. This elevation of [Ca 2+] i depended on extracellular Ca 2+ but was reduced by the IP 3 receptor blocker, 2-APB. This effect was not due to a Ca 2+ release triggered by Ca 2+ entry. Immunostaining showed IP 3-receptors (type1-3) at the cellular membrane of HeLa-S3 cells, but not in U2-OS cells. Electrophysiological experiments showed an increased membrane conductance with cisplatin only when Ca 2+ was present extracellularly. Increase of [Ca 2+] i was related to the activation of calpain but not caspase-8 and triggered apoptosis in HeLa-S3 but not in U2-OS cells. Conclusions and implications: Our observations on the activation of IP 3-receptors, calcium entry and apoptotic rate by cisplatin in specific carcinogenic cells might open new possibilities in the treatment of some forms of cancer.
AB - Background and purpose: Cisplatin drives specific types of tumour cells to apoptosis. In this study we investigate the involvement of intracellular calcium ([Ca 2+] i) in triggering apoptosis in two different cell lines. As cisplatin is used for the treatment of several forms of cancer we choose HeLa-S3 and U2-OS as two examples of tumour cell lines. Experimental approach: Cisplatin (1nM-10μM) was applied to HeLa-S3 and U2-OS cells and [Ca 2+] i measured with fluo-4, using laser scanning microscopy. Inositol-1,4,5-trisphosphate (IP 3) receptors were visualized with immunostaining. Membrane conductances were measured with patch-clamp techniques. Levels of calpain and caspases were assessed by western blots and apoptotic cells were stained with Hoechst 33342 and counted. Key results: Cisplatin increases [Ca 2+] i concentration- dependently in HeLa-S3 but not in U2-OS cells. This elevation of [Ca 2+] i depended on extracellular Ca 2+ but was reduced by the IP 3 receptor blocker, 2-APB. This effect was not due to a Ca 2+ release triggered by Ca 2+ entry. Immunostaining showed IP 3-receptors (type1-3) at the cellular membrane of HeLa-S3 cells, but not in U2-OS cells. Electrophysiological experiments showed an increased membrane conductance with cisplatin only when Ca 2+ was present extracellularly. Increase of [Ca 2+] i was related to the activation of calpain but not caspase-8 and triggered apoptosis in HeLa-S3 but not in U2-OS cells. Conclusions and implications: Our observations on the activation of IP 3-receptors, calcium entry and apoptotic rate by cisplatin in specific carcinogenic cells might open new possibilities in the treatment of some forms of cancer.
KW - [Ca ]
KW - Anticancer drugs
KW - Apoptosis
KW - Calcium signalling
KW - Calcium stores
KW - Calpain
KW - Carboplatin
KW - Cisplatin
KW - HeLa-S3
KW - IP -receptor
KW - U2-OS
UR - http://www.scopus.com/inward/record.url?scp=34547870107&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34547870107&partnerID=8YFLogxK
U2 - 10.1038/sj.bjp.0707335
DO - 10.1038/sj.bjp.0707335
M3 - Article
VL - 151
SP - 1176
EP - 1186
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
SN - 0007-1188
IS - 8
ER -