Activation of a Ca2+-permeable cation channel by two different inducers of apoptosis in a human prostatic cancer cell line

1. We have combined patch clamp recording with simultaneous [Ca2+](i) measurements in single LNCaP cells (a human prostate cancer cell line), to study the activation of Ca2+-permeable channels by two different inducers of apoptosis, ionomycin and serum deprivation. 2. In perforated patch recording, LNCaP cells had a membrane potential of -40 mV and a resting [Ca2+](i). The first rise in [Ca2+](i) was due to release of Ca2+ from internal stores and it was associated with a membrane hyperpolarization to -77 mV. The latter was probably due to the activation of high conductance, Ca2+- and voltage-dependent K+ channels (maxi-K). Conversely, the second rise in [Ca2+](i) was always preceded by and strictly associated with membrane depolarization and required external Ca2+. Serum deprivation, another inducer of apoptosis, unmasked a voltage-independent Ca2+ permeability as well. 3. A lower concentration of ionomycin (12 mu M) did not induce apoptosis, and neither depolarized LNCaP cells nor produced the biphasic increase in [Ca2+](i). However, the first increment in [Ca2+](i) due to release from internal Ca2+ stores was evident at this concentration of ionomycin. 4. Simultaneous recordings of [Ca2+](i) and ion channel activity in the cell attached configuration of patch clamp revealed a Ca2+-permeable, Ca2+-independent, non-selective cation channel of 23 pS conductance. This channel was activated only during the second increment in [Ca2+](i) induced by ionomycin. The absence of serum activated the 23 pS channel as well, albeit at a lower frequency than with ionomycin. 5. thus, the 23 pS channel can be activated by two unrelated inducers of apoptosis and it could be another Ca2+ influx mechanism in programmed cell death of LNCaP cells.