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Cellular, Molecular, and Tumor Biology 89: Mouse Models of Prostate and Gastrointestinal Cancers

Abstract #4319

Photodynamic therapy (PDT) with Hypericum perforatum L extract induced significant in vitro cytotoxicity of human bladder cancer cells

Virginia Commonwealth University, Richmond, VA, G.Hatzikosta General Hospital, Ioannina, Greece, University of Ioannina, Ioannina, Greece

INTRODUCTION AND OBJECTIVE: Hypericum perforatum L. (St. John’s wort) is a medicinal plant used for many human ailments. Locally, the Greeks have traditionally used both externally and internally for therapeutic indications. The anti-inflammatory, anti-microbial, antiviral, and antidepressant activities of this herb, have been attributed to the total extract or single constituents. We have reported on the antiproliferative activity of the lipophilic extract of the Epirus’ Hypericum perforatum L. (HP) against cultured T24, and NBT-11 bladder cancer cells. We investigated the efficacy of the polar methanolic fraction (PMF) of the HP extract in photodynamic therapy (PDT) alone, and in combination with recombinant Interferon-2b and Gemcitabine in the treatment of human TCC lines. METHODS: The polar methanolic fraction (PMF) was prepared after extraction of the dry herb with methanol, followed by liquid extraction with petroleum ether Human bladder tumor cell lines, T24, were plated (10⁴ cells/well) and placed in the incubator (37° C, 5%CO) for 24 hours prior to addition of drugs. PMF60ug/ml was added and incubation continued. After 24 hours, the cells were subjected to laser light (630nm) treatment with 0, 1, 4 and 8 Joules. After reincubation for 24 hours, Intron A (50,000 IU) or Gemcitabine (GTB, 2ug/ml) was added to the PDT-treated cells. After this incubation cell survival was assessed by the MTT assay. RESULTS: PDT with 60µg/ml of PMF alone induced percent cell kill of 9%, 8%,44% and 80% at 0,2,4 and 8 Joules respectively. Intron A (50,000 IU) alone induced 20% cell kill. Combination of PMF-PDT and Intron A induced cell kill of 31%, 64 and 86 % at 2, 4, 8 Joules respectively. GTB (2ug/ml) alone induced 53% cell kill, as well as synergistic effects with PMF-PDT for cell kill of 63%, 80% and 88% at 2,4 and 8 joules respectively. CONCLUSIONS: Data suggest that PMF may be an effective agent for photodynamic therapy. Intron A /GTB and PDT achieved improved kill of bladder cancer cells in cultures. Confirmation of these results in preclinical studies may lead to clinical trials.