Skin
phototoxicity is one of the main side effects of
photodynamic therapy (
PDT). To overcome this problem, some new
photosensitizers have been developed with longer absorbance wavelengths and shorter half-life in the body. In this study, we investigated the mechanism of
PDT mediated by a new
chlorophyll derivative
photosensitizer,
9-hydroxypheophorbide alpha (9-HPbD), on AMC-HN-3
cancer cells.
Phototoxicity and apoptosis on AMC-HN-3 cells induced by 9-HPbD was exhibited in a time- and dose-dependent manner. Mitochondria and endoplasmic reticulum (ER) were observed as preferential sites of 9-HPbD accumulation. Photoactivation of 9-HPbD-loaded AMC-HN-3 cells led to a rapid generation of
reactive oxygen species (ROS) at 30 min, followed by a loss of mitochondrial membrane potential (
MMP) at 2 h, translocation of
apoptosis-inducing factor (AIF) at 2 h, and the release of
cytochrome c at 3 h following
PDT.
Caspase-12, an important
caspase involved in ER-induced apoptosis, and
C/EBP homologous protein (CHOP), an ER stress inducible
transcription factor, were also upregulated after
PDT (3-12 h and 6-12 h, respectively). Subsequently, activation of
caspase-9 at 6 h,
caspase-3 and PARP at 12 h also occurred in
PDT-treated AMC-HN-3 cells. The above observations demonstrate that both mitochondria and ER serve not only as the sites of sensitizer binding, but also the subcellular targets of 9-HPbD-PDT, effective activation of which is responsible for 9-HPbD
PDT-induced apoptosis in AMC-HN-3 cells.