UVA/B-induced apoptosis in human melanocytes involves translocation of cathepsins and Bcl-2 family members.
We demonstrate UVA/B to induce apoptosis in human melanocytes through the mitochondrial pathway, displaying cytochrome c release, caspase-3 activation, and fragmentation of nuclei. The outcome of a death signal depends on the balance between positive and negative apoptotic regulators, such as members of the Bcl-2 protein family. Apoptotic melanocytes, containing fragmented nucleus, show translocation of the proapoptotic proteins Bax and Bid from the cytosol to punctate mitochondrial-like structures. Bcl-2, generally thought to be attached only to membranes, was in melanocytes localized in the cytosol as well. In the fraction of surviving melanocytes, that is, cells with morphologically unchanged nucleus, the antiapoptotic proteins Bcl-2 and Bcl-X(L) were translocated to mitochondria following UVA/B. The lysosomal proteases, cathepsin B and D, which may act as proapoptotic mediators, were released from lysosomes to the cytosol after UVA/B exposure. Proapoptotic action of the cytosolic cathepsins was confirmed by microinjection of cathepsin B, which induced nuclear fragmentation. Bax translocation and apoptosis were markedly reduced in melanocytes after pretreatment with either cysteine or aspartic cathepsin inhibitors. No initial caspase-8 activity was detected, excluding involvement of the death receptor pathway. Altogether, our results emphasize translocation of Bcl-2 family proteins to have central regulatory functions of UV-induced apoptosis in melanocytes and suggest cathepsins to be proapoptotic mediators operating upstream of Bax.
Bivik
Cecilia A
CA
Division of Dermatology, Faculty of Health Sciences, Linköping University, Linköping S-581 85, Sweden. cecbi@ibk.liu.se
Larsson
Petra K
PK
Kågedal
Katarina M
KM
Rosdahl
Inger K
IK
Ollinger
Karin M
KM
D017209:Apoptosis / Q000528:radiation effects*; D053181:Caspase 8; D020169:Caspases / Q000378:metabolism; D002403:Cathepsins / Q000502:physiology*; D002478:Cells, Cultured; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008544:Melanocytes / Q000528:radiation effects*; D021381:Protein Transport; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D014466:Ultraviolet Rays; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D051020:bcl-X Protein / Q000378:metabolism
D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D002403:Cathepsins; C505530:CASP8 protein, human; D053181:Caspase 8; D020169:Caspases