p53 Status does not affect photodynamic cell killing induced by hypericin.
OBJECTIVE
Given that p53 is a tumor suppressor that plays a central role in the cellular response to DNA damage and that more than 50% of all cancers have mutated p53, the wider utility of photodynamic therapy (PDT) in the treatment of cancer will depend on an understanding of whether p53 status modulates response to PDT. In this study, we investigated the photosensitivity of isogenic cell lines that differ only in their p53 status to PDT using hypericin as the photosensitizer.


METHODS
Acute (MTT) and chronic (clonogenic) cytotoxic assays were performed on two osteosarcoma cell-lines (U2OS and U2OS+p53DD) that are isogenic except that the latter expresses dominant negative p53. The inducible expression of p53 was determined on western blots. Uptake of hypericin, cell cycle profile analysis, measurement of membrane phosphatidylserine externalization and changes in mitochondrial membrane potential were investigated using flow cytometry.


RESULTS
Hypericin uptake was observed to be equivalent in U2OS and U2OS+p53DD cells. There were no significant differences in cell killing between these cell-lines in both the MTT and clonogenic assays (IC(50) of 0.4 microg/ml from MTT assay). p53 expression did not increase up to 24 h after PDT treatment in both cell lines. There were also no significant differences in the cell-cycle arrest profiles and timing of onset of apoptosis.


CONCLUSIONS
Taken together, these results suggest that the status of p53 may not be important in PDT-mediated cell killing or induction of apoptosis. By extension, these results imply that PDT may be used with equal efficacy for the treatment of p53-positive and -negative tumors.
Lee
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HB
Cancer Research Initiatives Foundation, CARIF, Subang Jaya Medical Centre, 1 Jalan SS12/1A, 47500 Subang Jaya, Selangor, Malaysia.
Ho
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AS
Teo
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SH
D000873:Anthracenes; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D012516:Osteosarcoma / Q000188:drug therapy / Q000378:metabolism; D010569:Perylene / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D010778:Photochemotherapy; D011838:Radiation-Sensitizing Agents / Q000493:pharmacokinetics / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
D000873:Anthracenes; D011838:Radiation-Sensitizing Agents; D016159:Tumor Suppressor Protein p53; D010569:Perylene; C004965:hypericin