Photodynamic effects of the cationic porphyrin, mesotetra(4N-methylpyridyl)porphine, on microtubules of HeLa cells.

The treatment of HeLa human carcinoma cells with mesotetra(4N-methylpyridyl)porphine (T4MPyP) and blue light led to damage of the microtubules (MTs). The morphologies of interphase MTs and the mitotic spindle apparatus were analysed by immunofluorescence staining of alpha-tubulin. The extent of MT damage depended on the light dose and the time after photodynamic treatment. After a period of 1 h after irradiation with doses of 0.3 or 1.5 J cm-2 (sublethal conditions, corresponding to survival rates of 90% and 60% respectively), the normal MT network arrangement of interphase cells and the mitotic spindle apparatus of many cells were clearly affected. However, these effects were found to be transient, and several hours after irradiation most MTs resumed control morphology. Higher irradiation doses (4.5 J cm-2, lethal conditions, less than 10% cell survival) resulted in the irreversible alteration of interphase and mitotic MTs. The change in MT organization appeared to be the reason for the observed increase in the mitotic index (MI) after sublethal doses. The largest increase in MI was detected 6 h after treatment (twofold increase over untreated cells) for both sublethal light doses. Most of the cells in mitosis corresponded to metaphase, the number of ana-telophase cells being greatly reduced for the first hours after irradiation with a dose of 1.5 J cm-2. The results, which resemble those observed with inhibitors of MT assembly, suggest that MTs might represent an important target for T4MPyP action.