Reorganization of cytoskeleton induced by 5-aminolevulinic acid-mediated photodynamic therapy and its correlation with mitochondrial dysfunction.
OBJECTIVE
This study investigated the early cellular events which occurred after mitochondrial photodamage induced by 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT).


METHODS
Subcellular localization of protoporphyrin IX (PpIX) in NIH3T3 cells was studied by confocal microscopy. Mitochondrial damage was assessed by measuring mitochondrial transmembrane potential and ATP contents, and confirmed by characteristic appearance on transmission electron microscopy. Cellular adhesion was measured by the level of resistance to trypsinization. Cytoskeletal studies were performed by fluorescent staining of cytoskeletal components.


RESULTS
Following ALA-PDT, mitochondrial damage was found in NIH3T3 cells as judged by the decrease of membrane potential and ATP contents. Mitochondrial photodamage was further confirmed by electron microscopy. Resistance to trypsinization after ALA-PDT was shown to be light dose-dependent. The increase of cellular adhesion after ALA-PDT was correlated with mitochondrial photodamage and reorganization of cytoskeletal components in NIH3T3 cells.


CONCLUSIONS
This study has demonstrated that mitochondrial dysfunctions induced by ALA-PDT results in alterations of cellular morphology and cellular adhesion.
Tsai
Jui-Chang
JC
Department of Surgery, Division of Neurosurgery, National Taiwan University Hospital, Taipei 100, Taiwan.
Wu
Chia-Lun
CL
Chien
Hsiung-Fei
HF
Chen
Chin-Tin
CT
D000135:Acid Phosphatase / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000622:Aminolevulinic Acid / Q000494:pharmacology*; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000528:radiation effects; D003599:Cytoskeleton / Q000187:drug effects* / Q000473:pathology / Q000528:radiation effects; D008564:Membrane Potentials; D051379:Mice; D046529:Microscopy, Electron, Transmission; D018715:Microscopy, Video; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D041681:NIH 3T3 Cells / Q000187:drug effects* / Q000528:radiation effects; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011524:Protoporphyrins / Q000378:metabolism
D017319:Photosensitizing Agents; D011524:Protoporphyrins; D000622:Aminolevulinic Acid; D000255:Adenosine Triphosphate; C028025:protoporphyrin IX; D000135:Acid Phosphatase