Photosensitizing properties of
5,10,15,20-tetrakis(4-hydroxyphenyl)porphyrin (
p-THPP) functionalized by covalent attachment of one chain of poly(
ethylene glycol) (PEG) with a molecular weight of 350, 2000, or 5000 Da (p-THPP-PEG(350), p-THPP-PEG(2000), p-THPP-PEG(5000)) were studied in vitro. Dark and photo cytotoxicity of these
photosensitizers delivered in
solution or embedded in
liposomes were evaluated on two cell lines: a human
colorectal carcinoma cell line (HCT 116) and a
prostate cancer cell line (DU 145), and compared with these treated with free
p-THPP. The attachment of PEG chains results in the pronounced reduction of the dark cytotoxicity of the parent
porphyrin. Cell viability tests have demonstrated that the
phototoxicity of pegylated
porphyrins is dependent on the length of PEG chain and p-THPP-PEG(2000) exhibited the highest photodynamic efficacy for both cell lines. The encapsulation into
liposomes did not improve the
PDT effect. However, the liposomal formulation of p-THPP-PEG(2000) showed a greater tendency to induce apoptosis in both cell lines than the parent or pegylated
porphyrin delivered in
solution. The colocalization of
p-THPP, p-THPP-PEG(2000) and p-THPP-PEG(2000) enclosed in
liposomes with fluorescent markers for lysosomes, mitochondria, endoplasmatic reticulum (ER) and Golgi apparatus (GA) was determined in the HCT 116 line. The
p-THPP exhibited ubiquitous intracellular distribution with a preference for membranes: mitochondria, ER, GA, lysosomes and plasma membrane. Fluorescence of p-THPP-PEG(2000) was observed within the cytoplasm, with a stronger signal detected in membranous organelle: mitochondria, ER, GA and lysosomes. In contrast, p-THPP-PEG(2000) delivered in
liposomes gave a distinct lysosomal pattern of localization.