ClC-3 chloride channel prevents apoptosis induced by thapsigargin in PC12 cells.
Cell volume can be altered by two different ways, swelling and shrinkage. Cell swelling is regulated by volume-regulated Cl- channel (VRC). It is not well understood whether shrinkage is regulated by VRC. We previously found that antisense oligonucleotide specific to ClC-3 (ClC-3 antisense) prevented cell proliferation, which was related to cell swell volume regulation. In the present study, we further studied the role of ClC-3 Cl- channel in cell apoptosis which was related to cell shrinkage volume regulation by using antisense oligonucleotide specific to ClC-3 (ClC-3 antisense) and ClC-3 cDNA transfection techniques. We found that thapsigargin (TG), a specific inhibitor of the endoplasmic reticulum calcium ATPase, evoked apoptotic morphological changes (including cytoplasmic blebbing, condensation of nuclear chromatin, and the formation of apoptotic bodies), DNA laddering, and caspase-3 activation in PC12 cells (Pheochromocytoma-derived cell line). TG increased the cell apoptotic population with a decrease in cell viability. These effects were consistent with the decrease in endogenous ClC-3 protein expression, which was also induced by TG. Overexpression of ClC-3 significantly inhibited TG effect on PC12 cell apoptosis, whereas the ClC-3 antisense produced opposite effects and facilitated apoptosis induced by TG. Our data strongly suggest that ClC-3 channel in PC12 cells mediates TG-induced apoptotic process through inhibitory mechanism. Thus, it appears that ClC-3 Cl- channel mediates both cell proliferation and apoptosis through accelerative and inhibitory fashions, respectively.
Zhang
H-N
HN
Department of Pharmacology, Zhongshan Medical College, Sun Yat-Sen University, 74 Zhongshan 2 Road, Guangzhou, Guangdong, 510089, People's Republic of China.
Zhou
J-G
JG
Qiu
Q-Y
QY
Ren
J-L
JL
Guan
Y-Y
YY
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D048430:Cell Shape; D048429:Cell Size; D018118:Chloride Channels / Q000235:genetics / Q000378:metabolism*; D053938:DNA Fragmentation; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism; D016716:PC12 Cells; D051381:Rats; D019284:Thapsigargin / Q000378:metabolism*
D018118:Chloride Channels; C115376:ClC-3 channel; D004791:Enzyme Inhibitors; D016376:Oligonucleotides, Antisense; D019284:Thapsigargin; D053148:Caspase 3