title: The diverse roles of glutathione-associated cell resistance against hypericin photodynamic therapy.
abstract: The diverse responses of different cancers to treatments such as photodynamic therapy of cancer (PDT) have fueled a growing need for reliable predictive markers for treatment outcome. In the present work we have studied the differential response of two phenotypically and genotypically different breast adenocarcinoma cell lines, MCF7 and MDA-MB-231, to hypericin PDT (HYP-PDT). MDA-MB-231 cells were 70% more sensitive to HYP PDT than MCF7 cells at LD50. MCF7 were found to express a substantially higher level of glutathione peroxidase (GPX4) than MDA-MB-231, while MDA-MB-231 differentially expressed glutathione-S-transferase (GSTP1), mainly used for xenobiotic detoxification. Eighty % reduction of intracellular glutathione (GSH) by buthionine sulfoximine (BSO), largely enhanced the sensitivity of the GSTP1 expressing MDA-MB-231 cells to HYP-PDT, but not in MCF7 cells. Further inhibition of the GSH reduction however by carmustine (BCNU) resulted in an enhanced sensitivity of MCF7 to HYP-PDT. HYP loading studies suggested that HYP can be a substrate of GSTP for GSH conjugation as BSO enhanced the cellular HYP accumulation by 20% in MDA-MB-231 cells, but not in MCF7 cells. Studies in solutions showed that L-cysteine can bind the GSTP substrate CDNB in the absence of GSTP. This means that the GSTP-lacking MCF7 may use L-cysteine for xenobiotic detoxification, especially during GSH synthesis inhibition, which leads to L-cysteine build-up. This was confirmed by the lowered accumulation of HYP in both cell lines in the presence of BSO and the L-cysteine source NAC. NAC reduced the sensitivity of MCF7, but not MDA-MB-231, cells to HYP PDT which is in accordance with the antioxidant effects of L-cysteine and its potential as a GSTP substrate. As a conclusion we have herein shown that the different GSH based cell defense mechanisms can be utilized as predictive markers for the outcome of PDT and as a guide for selecting optimal combination strategies.
authors.affiliation: Department of Radiation Biology, Institute for cancer Research, Radium Hospital, Oslo University Hospital, Montebello, 0379 Oslo, Norway. Electronic address: Theodossis.Theodossiou@rr-research.no.
authors.forename: Theodossis A
authors.initials: TA
authors.lastname: Theodossiou
authors.affiliation: Department of Radiation Biology, Institute for cancer Research, Radium Hospital, Oslo University Hospital, Montebello, 0379 Oslo, Norway.
authors.forename: Cathrine E
authors.initials: CE
authors.lastname: Olsen
authors.affiliation: Department of Radiation Biology, Institute for cancer Research, Radium Hospital, Oslo University Hospital, Montebello, 0379 Oslo, Norway.
authors.forename: Marte
authors.initials: M
authors.lastname: Jonsson
authors.affiliation: PLANTA Naturstoffe Vertriebs GmbH, A-1120 Wien, Austria.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kubin
authors.affiliation: Department of Radiation Biology, Institute for cancer Research, Radium Hospital, Oslo University Hospital, Montebello, 0379 Oslo, Norway.
authors.forename: John S
authors.initials: JS
authors.lastname: Hothersall
authors.affiliation: Department of Radiation Biology, Institute for cancer Research, Radium Hospital, Oslo University Hospital, Montebello, 0379 Oslo, Norway.
authors.forename: Kristian
authors.initials: K
authors.lastname: Berg
mesh_terms: D000873:Anthracenes; D001943:Breast Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002330:Carmustine / Q000494:pharmacology; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm; D005260:Female; D005978:Glutathione / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000378:metabolism; D051549:Glutathione S-Transferase pi / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D010569:Perylene / Q000031:analogs & derivatives* / Q000494:pharmacology; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D010778:Photochemotherapy
chemical_list: D000873:Anthracenes; D019328:Buthionine Sulfoximine; D010569:Perylene; C004965:hypericin; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D005979:Glutathione Peroxidase; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005978:Glutathione; D002330:Carmustine
keywords: Glutathione; Glutathione peroxidases; Glutathione-S- transferases; Hypericin; Photodynamic therapy of Cancer; Predictive markers for PDT