Deregulation of apoptotic volume decrease and ionic movements in multidrug-resistant tumor cells: role of chloride channels

Abstract

Changes in cell volume and ion gradients across the plasma membrane play a pivotal role in the initiation of apoptosis. Here we explore the kinetics of apoptotic volume decrease (AVD) and ion content dynamics in wild-type (WT) and multidrug-resistant (MDR) Ehrlich ascites tumor cells (EATC). In WT EATC, induction of apoptosis with cisplatin (5 μM) leads to three distinctive AVD stages: an early AVD1 (4–12 h), associated with a 30% cell water loss; a transition stage AVDT (∼12 to 32 h), where cell volume is partly recovered; and a secondary AVD2 (past 32 h), where cell volume was further reduced. AVD1 and AVD2 were coupled to net loss of Cl−, K+, Na+, and amino acids (ninhydrin-positive substances), whereas during AVDT, Na+ and Cl− were accumulated. MDR EATC was resistant to cisplatin, showing increased viability and less caspase 3 activation. Compared with WT EATC, MDR EATC underwent a less pronounced AVD1, an augmented AVDT, and a delay in induction of AVD2. Changes in AVD were associated with inhibition of Cl− loss during AVD1, augmented NaCl uptake during AVDT, and a delay of Cl− loss during AVD2. Application of the anion channel inhibitor NS3728 inhibited AVD and completely abolished the differences in AVD, ionic movements, and caspase 3 activation between WT and MDR EATC. Finally, the maximal capacity of volume-regulated anion channel was found to be strongly repressed in MDR EATC. Together, these data suggest that impairment of AVD, primarily via modulation of NaCl movements, contribute to protection against apoptosis in MDR EATC.