Mitochondrial localization and photodamage during photodynamic therapy with tetraphenylporphines.
The subcellular localization sites of TPPS4 and TPPS1 and the subsequent cellular site damage during photodynamic therapy were investigated in CT-26 colon carcinoma cells using spectroscopic and electron microscopy techniques. The association of both porphyrins with the mitochondria was investigated and the implications of this association on cellular functions were determined. Spectrofluorescence measurements showed that TPPS4 favors an aqueous environment, while TPPS1 interacts with lipophilic complexes. The subcellular localization sites of each sensitizer were determined using spectral imaging. Mitochondrial-CFP transfected cells treated with porphyrins revealed localization of TPPS1 in the peri-nuclear region, while TPPS4 localized in the mitochondria, inducing structural damage and swelling upon irradiation, as shown by transmission electron microscopy. TPPS4 fluorescence was detected in isolated mitochondria following irradiation. The photodamage induced a 38% reduction in mitochondrial activity, a 30% decrease in cellular ATP and a reduction in Na(+)/K(+)-ATPase activity. As a result, cytosolic concentrations of Na(+) and Ca(2+) increased, and the level of K(+) decreased. In contrast, the lipophilic TPPS1 did not affect mitochondrial structure or function and ATP content remained unchanged. We conclude that TPPS4 induces mitochondrial structural and functional photodamage resulting in an altered cytoplasmic ion concentration, while TPPS1 has no effect on the mitochondria.
Weizman
E
E
Microscopy Unit, Faculty of Life Sciences, Bar-llan University, Ramat-Gan, Israel.
Rothmann
C
C
Greenbaum
L
L
Shainberg
A
A
Adamek
M
M
Ehrenberg
B
B
Malik
Z
Z
D000230:Adenocarcinoma; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003110:Colonic Neoplasms; D007202:Indicators and Reagents / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D008928:Mitochondria / Q000378:metabolism*; D010778:Photochemotherapy; D011166:Porphyrins / Q000378:metabolism*; D011838:Radiation-Sensitizing Agents / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013778:Tetrazolium Salts / Q000378:metabolism; D014407:Tumor Cells, Cultured
D007202:Indicators and Reagents; D011166:Porphyrins; D011838:Radiation-Sensitizing Agents; D013778:Tetrazolium Salts; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; C016136:tetraphenylporphine sulfonate; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase