Photocytotoxic and DNA damaging effect of Temoporfin (mTHPC) and merocyanine 540 (MC540) on nasopharyngeal carcinoma cell

Abstract

Photodynamic therapy (PDT) is a new approach to cancer treatment for a variety of malignant tumors. In this study, two clinical photosensitizers, Temoporfin (meta-tetra-hydroxyl-phenyl-chlorin; mTHPC) and merocyanine 540 (MC540), were selected to explore for their photocytotoxic and genotoxic effects on nasopharyngeal carcinoma cells (NPC/HK1 and CNE2). Results of tetrazolium reduction assay showed that 80% cell killing were achieved for both cell lines at 0.4 μg/ml mTHPC for 24 h incubation and then with 40 kJ/m 2 light irradiation, whereas 40 μg/ml MC540 with 50 kJ/m 2 light dosage was required to attain the same level of phototoxicity for NPC/HK1. On the contrary, NPC/CNE2 was quite resistant to MC540. Hence, mTHPC-mediated PDT exerted a more potent effect than MC540-mediated PDT, even though the molar extinction coefficient of the main absorption peak for MC540 is much higher than that of mTHPC. Dark cytotoxicity remained negligible for both sensitizers. Comet assay was used to evaluate the DNA strand break and potential genotoxic effect induced by mTHPC and MC540 on the NPC cells. No DNA strand break was detected in the absence of light, and under sublethal treatment (LD 25) for either sensitizer-loaded cells. Confocal laser scanning microscopy showed that mTHPC and MC540 localized in the cytoplasm but not in the nucleus of the tumor cells, which provided evidence for undetectable DNA damage under dark and low photodynamic dose.