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Romidepsin (FK228) combined with cisplatin stimulates DNA damage-induced cell death in ovarian cancer.

ABSTRACT: Romidepsin (FK228) was recently approved by the FDA for the treatment of cutaneous and peripheral T cell lymphoma. We have shown in vitro efficacy of FK228 in ovarian cancer. Here, our goal was to evaluate FK228 combined with cisplatin in ovarian cancer in vitro and in vivo.Ovarian cancer cell lines were treated with cisplatin, FK228 or the combination of drugs. Colorimetric assays were used to determine cytotoxicity in vitro. Mice engrafted with 5×10(6) SKOV-3 ovarian cancer cells were treated with cisplatin, FK228 or the combination, and tumor weights and volumes were measured. We assessed molecular markers of proliferation (mib-1), apoptosis (cleaved PARP and cleaved caspase 3) and DNA damage (pH2AX, RAD51 and 53BP1).FK228 enhanced the cytotoxic effects of cisplatin in ovarian cells compared to vehicle-treated controls or each drug alone. The combination of FK228 and cisplatin-induced apoptosis and activated aberrant DNA damage responses demonstrated by increased expression of pH2AX, RAD51 and 53BP1. Mice treated with FK228, cisplatin and both drugs showed reduced tumor weights and volumes. Drug-treated tumors showed decreased mib-1 and increased cleaved-caspase 3 expression levels. The number and intensity of pH2AX stained cells was greatest in tumors exposed to the combination of FK228 and cisplatin.FK228 causes DNA damage-induced apoptosis and enhances the anti-tumor effects of cisplatin. The DNA damage mark pH2AX is activated by FK228 and cisplatin and may be a useful pharmacodynamic mark of these effects.

SUBMITTER: Wilson AJ

PROVIDER: S-EPMC3541411 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature

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Romidepsin (FK228) combined with cisplatin stimulates DNA damage-induced cell death in ovarian cancer.

Wilson Andrew J AJ Lalani Anum S AS Wass Erika E Saskowski Jeanette J Khabele Dineo D

Gynecologic oncology 20120923 3

<h4>Objective</h4>Romidepsin (FK228) was recently approved by the FDA for the treatment of cutaneous and peripheral T cell lymphoma. We have shown in vitro efficacy of FK228 in ovarian cancer. Here, our goal was to evaluate FK228 combined with cisplatin in ovarian cancer in vitro and in vivo.<h4>Methods</h4>Ovarian cancer cell lines were treated with cisplatin, FK228 or the combination of drugs. Colorimetric assays were used to determine cytotoxicity in vitro. Mice engrafted with 5×10(6) SKOV-3 ...[more]

PMID: 23010348

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Project description:BackgroundCisplatin (CDDP), a widely used chemotherapeutic agent, can induce excessive granulosa cell apoptosis, follicle loss and even premature ovarian insufficiency (POI). However, the mechanism remains elusive, although some studies have indicated the involvement of endoplasmic reticulum stress (ERS). The aim of our study was to investigate the possible mechanism ERS in CDDP-induced granulosa cell apoptosis and follicle loss.MethodsA POI mouse model was generated by CDDP. The ovaries samples were collected and processed for isobaric tags for relative and absolute quantification analysis (iTRAQ) to screen out our interested proteins of HSPA5 and HSP90AB1, and the decline in their expression were verified by a real-time quantitative PCR and a western blotting assay. In vitro, human granulosa cells, KGN and COV434 cells were transfected with siRNA targeting HSPA5 and HSP90AB1 and then treated with CDDP, or treated with CDDP with/without CDDP+?4-phenylbutyric acid (4-PBA) and 3-methyladenine (3-MA). The levels of ERS, autophagy and apoptosis were evaluated by western blotting, DALGreen staining and flow cytometry. In vivo, ovaries from mice that received intraperitoneal injections of saline, CDDP, CDDP+?4-PBA and CDDP+?3-MA were assayed by immunofluorescence, hematoxylin and eosin (H&E) staining for follicle counting, and terminal-deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) staining for cell apoptosis assay. The plasma hormone levels were measured by an enzyme-linked immunosorbent assay (ELISA) kit.ResultsWe have clarified the relationships between ERS, autophagy, and apoptosis in CDDP-induced granulosa cell apoptosis, both in vitro and in vivo. Alleviating ERS by inhibiting HSPA5 and HSP90AB1 attenuated CDDP-induced autophagy and apoptosis. 4-PBA treatment significantly attenuated CDDP-induced cell autophagy and apoptosis in cultured KGN and COV434 cells. However, inhibiting cell autophagy with 3-MA negligibly restored the CDDP-induced changes in ERS and apoptosis. In vivo experiments also demonstrated that treatment with 4-PBA, but not 3-MA, prevented CDDP-induced ovarian damage and hormone dysregulation.ConclusionsCDDP-induced ERS could promote autophagy and apoptosis in granulosa cells, causing excessive follicle loss and endocrine disorders. Alleviation of ERS with 4-PBA, but not of autophagy with 3-MA, protect against CDDP-induced granulosa cell apoptosis and ovarian damage. Thus, 4-PBA can be used to protect the ovary during chemotherapy in women.

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Romidepsin (FK228) combined with cisplatin stimulates DNA damage-induced cell death in ovarian cancer.

Publications

Romidepsin (FK228) combined with cisplatin stimulates DNA damage-induced cell death in ovarian cancer.

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