Abstract
Charybdotoxin, a blocker of K + channels, and the imidazole drug SC38249, a blocker of both voltage- and second messenger-operated Ca 2+ channels, were employed in mouse NIH-3T3 fibroblasts overexpressing the epidermal growth factor (EGF) receptor 1) to characterize the ionic events activated by EGF; and 2) to establish the role of those events in cell growth. The [Ca 2+](i) response by EGF was little changed by charybdotoxin while the parallel hyperpolarization was inhibited in a dose-dependent manner. At high toxin concentrations (>3 x 10 -8 M), the effect of EGF on membrane potential was turned into a persistent depolarization sustained by both Na + and Ca 2+. Pretreatment with 10 μM SC38249 induced only minor changes of the intracellular Ca 2+ release by EGF (the process responsible for the initial phase of the [Ca 2+](i) and membrane potential responses) and blocked the persistent, second phase [Ca 2+](i) and the hyperpolarization responses, both dependent on Ca 2+ influx, as well as the depolarization in the charybdotoxin-pretreated cells. Long term (up to 2-day) treatment with either charybdotoxin or SC38249 failed to affect the viability and growth of unstimulated EGFR-T17 cells. Moreover, in these cells, the ionic responses to EGF were restored after a 30-min incubation in fresh medium. In contrast, growth stimulated by EGF was inhibited, moderately (-20%) by charybdotoxin and markedly (-60%) by SC38249. These results indicate for the first time that both hyperpolarization and, especially, the persistent increase of [Ca 2+](i) sustained by Ca 2+ influx play a role in the activity of EGF, ultimately cooperating with other intracellular events in mitogenesis.
| Original language | English |
|---|---|
| Pages (from-to) | 6329-6335 |
| Number of pages | 7 |
| Journal | Journal of Biological Chemistry |
| Volume | 266 |
| Issue number | 10 |
| Publication status | Published - 1991 |
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ASJC Scopus subject areas
- Biochemistry
Cite this
Ionic events induced by epidermal growth factor : Evidence that hyperpolarization and stimulated cation influx play a role in the stimulation of cell growth. / Magni, M.; Meldolesi, J.; Pandiella, A.
In: Journal of Biological Chemistry, Vol. 266, No. 10, 1991, p. 6329-6335.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Ionic events induced by epidermal growth factor
T2 - Evidence that hyperpolarization and stimulated cation influx play a role in the stimulation of cell growth
AU - Magni, M.
AU - Meldolesi, J.
AU - Pandiella, A.
PY - 1991
Y1 - 1991
N2 - Charybdotoxin, a blocker of K + channels, and the imidazole drug SC38249, a blocker of both voltage- and second messenger-operated Ca 2+ channels, were employed in mouse NIH-3T3 fibroblasts overexpressing the epidermal growth factor (EGF) receptor 1) to characterize the ionic events activated by EGF; and 2) to establish the role of those events in cell growth. The [Ca 2+](i) response by EGF was little changed by charybdotoxin while the parallel hyperpolarization was inhibited in a dose-dependent manner. At high toxin concentrations (>3 x 10 -8 M), the effect of EGF on membrane potential was turned into a persistent depolarization sustained by both Na + and Ca 2+. Pretreatment with 10 μM SC38249 induced only minor changes of the intracellular Ca 2+ release by EGF (the process responsible for the initial phase of the [Ca 2+](i) and membrane potential responses) and blocked the persistent, second phase [Ca 2+](i) and the hyperpolarization responses, both dependent on Ca 2+ influx, as well as the depolarization in the charybdotoxin-pretreated cells. Long term (up to 2-day) treatment with either charybdotoxin or SC38249 failed to affect the viability and growth of unstimulated EGFR-T17 cells. Moreover, in these cells, the ionic responses to EGF were restored after a 30-min incubation in fresh medium. In contrast, growth stimulated by EGF was inhibited, moderately (-20%) by charybdotoxin and markedly (-60%) by SC38249. These results indicate for the first time that both hyperpolarization and, especially, the persistent increase of [Ca 2+](i) sustained by Ca 2+ influx play a role in the activity of EGF, ultimately cooperating with other intracellular events in mitogenesis.
AB - Charybdotoxin, a blocker of K + channels, and the imidazole drug SC38249, a blocker of both voltage- and second messenger-operated Ca 2+ channels, were employed in mouse NIH-3T3 fibroblasts overexpressing the epidermal growth factor (EGF) receptor 1) to characterize the ionic events activated by EGF; and 2) to establish the role of those events in cell growth. The [Ca 2+](i) response by EGF was little changed by charybdotoxin while the parallel hyperpolarization was inhibited in a dose-dependent manner. At high toxin concentrations (>3 x 10 -8 M), the effect of EGF on membrane potential was turned into a persistent depolarization sustained by both Na + and Ca 2+. Pretreatment with 10 μM SC38249 induced only minor changes of the intracellular Ca 2+ release by EGF (the process responsible for the initial phase of the [Ca 2+](i) and membrane potential responses) and blocked the persistent, second phase [Ca 2+](i) and the hyperpolarization responses, both dependent on Ca 2+ influx, as well as the depolarization in the charybdotoxin-pretreated cells. Long term (up to 2-day) treatment with either charybdotoxin or SC38249 failed to affect the viability and growth of unstimulated EGFR-T17 cells. Moreover, in these cells, the ionic responses to EGF were restored after a 30-min incubation in fresh medium. In contrast, growth stimulated by EGF was inhibited, moderately (-20%) by charybdotoxin and markedly (-60%) by SC38249. These results indicate for the first time that both hyperpolarization and, especially, the persistent increase of [Ca 2+](i) sustained by Ca 2+ influx play a role in the activity of EGF, ultimately cooperating with other intracellular events in mitogenesis.
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UR - http://www.scopus.com/inward/citedby.url?scp=0025865917&partnerID=8YFLogxK
M3 - Article
C2 - 1706715
AN - SCOPUS:0025865917
VL - 266
SP - 6329
EP - 6335
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 10
ER -