Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines
This study examined the effect of combination treatment with the poly (ADP-ribose) polymerase inhibitor olaparib and metformin on homologous recombination (HR)-proficient epithelial ovarian cancer (EOC). Ovarian cancer cell lines (OV-90 and SKOV-3) were treated with olaparib, metformin, or a combina...
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MDPI AG
2021-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/19/10557 |
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| author | Patrycja Gralewska Arkadiusz Gajek Agnieszka Marczak Aneta Rogalska |
| author_facet | Patrycja Gralewska Arkadiusz Gajek Agnieszka Marczak Aneta Rogalska |
| author_sort | Patrycja Gralewska |
| collection | DOAJ |
| description | This study examined the effect of combination treatment with the poly (ADP-ribose) polymerase inhibitor olaparib and metformin on homologous recombination (HR)-proficient epithelial ovarian cancer (EOC). Ovarian cancer cell lines (OV-90 and SKOV-3) were treated with olaparib, metformin, or a combination of both. Cell viability was assessed by MTT and colony formation assays. The production of reactive oxygen species (ROS) and changes in mitochondrial membrane potential were examined using the specific fluorescence probes, DCFH2-DA (2′,7′-dichloro-dihydrofluorescein diacetate) and JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine). Apoptotic and necrotic changes were measured by double staining with Hoechst 33258 and propidium iodide, orange acridine and ethidium bromide staining, phosphatidylserine externalization, TUNEL assay, caspase 3/7 activity, and cytochrome c and p53 expression. Compared with single-drug treatment, the combination of olaparib and metformin significantly inhibited cell proliferation and colony formation in HR-proficient ovarian cancer cells. ROS production preceded a decrease in mitochondrial membrane potential. The changes in ROS levels suggested their involvement in inducing apoptosis in response to combination treatment. The present results indicate a shift towards synergism in cells with mutant or null p53, treated with olaparib combined with metformin, providing a new approach to the treatment of gynecologic cancers. Taken together, the results support the use of metformin to sensitize EOC to olaparib therapy. |
| first_indexed | 2024-03-10T06:59:43Z |
| format | Article |
| id | doaj.art-3bad002c34544e5ab97d953d3707137b |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T06:59:43Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-3bad002c34544e5ab97d953d3707137b2023-11-22T16:11:46ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-09-0122191055710.3390/ijms221910557Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell LinesPatrycja Gralewska0Arkadiusz Gajek1Agnieszka Marczak2Aneta Rogalska3Department of Medical Biophysics, Faculty of Biology and Environmental Protection, Institute of Biophysics, University of Lodz, 90-236 Lodz, PolandDepartment of Medical Biophysics, Faculty of Biology and Environmental Protection, Institute of Biophysics, University of Lodz, 90-236 Lodz, PolandDepartment of Medical Biophysics, Faculty of Biology and Environmental Protection, Institute of Biophysics, University of Lodz, 90-236 Lodz, PolandDepartment of Medical Biophysics, Faculty of Biology and Environmental Protection, Institute of Biophysics, University of Lodz, 90-236 Lodz, PolandThis study examined the effect of combination treatment with the poly (ADP-ribose) polymerase inhibitor olaparib and metformin on homologous recombination (HR)-proficient epithelial ovarian cancer (EOC). Ovarian cancer cell lines (OV-90 and SKOV-3) were treated with olaparib, metformin, or a combination of both. Cell viability was assessed by MTT and colony formation assays. The production of reactive oxygen species (ROS) and changes in mitochondrial membrane potential were examined using the specific fluorescence probes, DCFH2-DA (2′,7′-dichloro-dihydrofluorescein diacetate) and JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine). Apoptotic and necrotic changes were measured by double staining with Hoechst 33258 and propidium iodide, orange acridine and ethidium bromide staining, phosphatidylserine externalization, TUNEL assay, caspase 3/7 activity, and cytochrome c and p53 expression. Compared with single-drug treatment, the combination of olaparib and metformin significantly inhibited cell proliferation and colony formation in HR-proficient ovarian cancer cells. ROS production preceded a decrease in mitochondrial membrane potential. The changes in ROS levels suggested their involvement in inducing apoptosis in response to combination treatment. The present results indicate a shift towards synergism in cells with mutant or null p53, treated with olaparib combined with metformin, providing a new approach to the treatment of gynecologic cancers. Taken together, the results support the use of metformin to sensitize EOC to olaparib therapy.https://www.mdpi.com/1422-0067/22/19/10557combination therapymetforminolaparibovarian cancerPARP inhibitorreplication stress |
| spellingShingle | Patrycja Gralewska Arkadiusz Gajek Agnieszka Marczak Aneta Rogalska Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines International Journal of Molecular Sciences combination therapy metformin olaparib ovarian cancer PARP inhibitor replication stress |
| title | Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines |
| title_full | Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines |
| title_fullStr | Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines |
| title_full_unstemmed | Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines |
| title_short | Metformin Affects Olaparib Sensitivity through Induction of Apoptosis in Epithelial Ovarian Cancer Cell Lines |
| title_sort | metformin affects olaparib sensitivity through induction of apoptosis in epithelial ovarian cancer cell lines |
| topic | combination therapy metformin olaparib ovarian cancer PARP inhibitor replication stress |
| url | https://www.mdpi.com/1422-0067/22/19/10557 |
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