C. Pourzand et al., The iron regulatory protein can determine the effectiveness of 5-aminolevulinic acid in inducing protoporphyrin IX in human primary skin fibroblasts, J INVES DER, 112(4), 1999, pp. 419-425
The level of endogenous photosensitiser, protoporphyrin IX (PPIX), can be e
nhanced in the cells by 5-aminolevulinic acid (ALA), We investigated the ef
fect of critical parameters such as growth state of the cells and availabil
ity of intracellular iron in modulating the level of PPIX, in human primary
cultured skin fibroblasts (FEK4) maintained either in exponentially growin
g or growth-arrested phase, following treatment with ALA. The addition of A
LA to exponentially growing cells increased the level of PPIX 6-fold relati
ve to control cells; however, in growth-arrested cells the same treatment i
ncreased the level of PPIX up to 34-fold. The simultaneous addition of the
hydrophilic iron-chelator Desferal with ALA, boosted the level of PPIX up t
o 47-fold in growing cells and up to 42-fold in growth-arrested cells, sugg
esting that iron is limiting under the latter conditions. The strict depend
ence of PPIX enhancement on free available iron levels was examined by the
level of activation of iron regulatory protein in band shift assays. This a
nalysis revealed that the basal level of iron regulatory protein in growth-
arrested cells was 6-fold higher than in growing cells, reflecting the infl
uence of the free available iron pool in exponentially growing cells, Inter
estingly, the same ratio was found between the basal level concentration of
PPIX in growing and growth-arrested cells. We propose that iron regulatory
protein activation could serve as a marker for developing photodynamic the
rapy protocols because it identifies cells and tissues with a propensity to
accumulate PPIX and it is therefore likely to predict the effectiveness of
such therapies.