Photo-oxidative action in cervix carcinoma cells induced by HPD - mediated photodynamic therapy.
Photodynamic therapy leads to oxidative stress through the generation of free radicals. Oxidative stress causes damage to cellular macromolecules such as nucleic acids, proteins and lipids.


OBJECTIVE
To examine the hematoporphyrin derivative (HpD) - mediated photodynamic effect on cervical adenocarcinoma cell line HeLa.


METHODS
The HpD localization in HeLa cells was analyzed by confocal microscopy with epi-fluorescence system. Lipid peroxidation (LPO) was estimated by measurement of the concentration of malondialdehyde, protein degradation - by modified Ellman's method, superoxide dysmutase (SOD) - using Ransod Kit. The expression of inducible nitric oxide synthase (iNOS) was detected by immunocytochemical staining.


RESULTS
The HpD was distributed all over the cytoplasm with preferential localization in the inner side of the plasma membrane and around the nuclear envelope. The process of photosensitizer distribution was time dependent. PDT-HpD increased the level of malonodialdehyde (MDA), SOD activity and the expression of iNOS in HeLa cells. However, PDT induced the decrease in the level of protein-associated thiol groups.


CONCLUSIONS
Our study showed the important role of PDT-mediated oxidative stress in HeLa cells. HpD-PDT might be alternative and less invasive approach for treatment of patients with cervical cancer resistant for standard chemotherapy and radiotherapy.
Saczko
J
J
Department of Medical Biochemistry, Silesian Piasts University of Medicine, 50-368 Wroclaw, Poland. michal@bioch.am.wroc.pl
Skrzypek
W
W
Chwiłkowska
A
A
Choromańska
A
A
Poła
A
A
Gamian
A
A
Kulbacka
J
J
D000230:Adenocarcinoma / Q000378:metabolism / Q000628:therapy*; D005260:Female; D006367:HeLa Cells; D017324:Hematoporphyrin Derivative / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D015227:Lipid Peroxidation / Q000187:drug effects; D018613:Microscopy, Confocal; D052247:Nitric Oxide Synthase Type II / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D010778:Photochemotherapy / Q000379:methods*; D013482:Superoxide Dismutase / Q000187:drug effects / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000628:therapy*
D017324:Hematoporphyrin Derivative; D052247:Nitric Oxide Synthase Type II; D013482:Superoxide Dismutase