Effects of photodynamic therapy on the endocytic pathway.
In this report, we describe an effect of photodynamic therapy (PDT) on membrane trafficking in murine 1c1c7 hepatoma cells. A brief exposure of 1c1c7 cells to a 20 nM concentration of the phosphatidylinositol kinase class-3 antagonist wortmannin led to the rapid appearance of cytoplasmic vacuoles. Fluorescence monitoring of plasma membrane-associated 1-[4-(trimethylamino)phenyl]-6-phenylhexa-1,3,5-triene (TDPH) over time demonstrated that the wortmannin-induced vacuoles were derived from endocytosed plasma membrane. Low-dose photodamage catalyzed by the lysosomal photosensitizer NPe6, prior to the addition of wortmannin, prevented formation of these vacuoles. NPe6 was found to suppress for several hours the normal trafficking of TDPH-labeled plasma membrane to the cytosol, and the formation of punctate TDPH-labeled cytoplasmic vesicles. The ability of NPe6-induced photodamage to suppress wortmannin-induced vacuolization occurred under conditions that did not disrupt lysosomes and were at or below the threshold of cytostatic/cytotoxic effects. Furthermore, the suppressive effects of NPe6-PDT were not prevented by inclusion of an agent that stabilized lysosomal membranes, or by E64d, an inhibitor of lysosomal cathepsin proteases. Mitochondrial photodamage was less effective at preventing wortmannin-induced vacuole formation and PDT directed against the ER had no effect. The role of photodamage to the endocytic pathway may be a hitherto unexplored effect on cells that selectively accumulate photosensitizing agents. These results indicate that photodamage directed against endosomes/lysosomes has effects independent of the release of lysosomal proteases.
Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
David
D
Kessel
Michael
M
Price
Joseph
J
Caruso
John
J
Reiners
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002403:Cathepsins / Q000037:antagonists & inhibitors / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D004161:Diphenylhexatriene / Q000031:analogs & derivatives / Q000737:chemistry; D004705:Endocytosis / Q000528:radiation effects*; D011992:Endosomes / Q000187:drug effects / Q000528:radiation effects; D005456:Fluorescent Dyes / Q000737:chemistry; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology / Q000528:radiation effects; D051379:Mice; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011166:Porphyrins / Q000633:toxicity; D013997:Time Factors; D014617:Vacuoles / Q000378:metabolism; D000077191:Wortmannin
D000730:Androstadienes; D005456:Fluorescent Dyes; D017319:Photosensitizing Agents; D011166:Porphyrins; D004161:Diphenylhexatriene; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D002403:Cathepsins; D007930:Leucine; C108192:aloxistatin; C053434:Talaporfin; D000077191:Wortmannin