Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK
Purpose: 3,3′-Diindolylmethane (DIM), derived from indole-3-carbinol (I3C) in the Brassica species of cruciferous vegetables, has anticancer effects, but its exact underlying mechanism of action is unknown. We explored the roles of cytosolic free calcium ([Ca2+]i) and p38 MAPK in the anti-cancer eff...
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Frontiers Media S.A.
2019-10-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphar.2019.01167/full |
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| author | Yuanyue Jiang Yuanyue Jiang Yanfei Fang Yanfei Fang Yang Ye Xinming Xu Bingfang Wang Jie Gu Michael Aschner Jian Chen Rongzhu Lu Rongzhu Lu |
| author_facet | Yuanyue Jiang Yuanyue Jiang Yanfei Fang Yanfei Fang Yang Ye Xinming Xu Bingfang Wang Jie Gu Michael Aschner Jian Chen Rongzhu Lu Rongzhu Lu |
| author_sort | Yuanyue Jiang |
| collection | DOAJ |
| description | Purpose: 3,3′-Diindolylmethane (DIM), derived from indole-3-carbinol (I3C) in the Brassica species of cruciferous vegetables, has anticancer effects, but its exact underlying mechanism of action is unknown. We explored the roles of cytosolic free calcium ([Ca2+]i) and p38 MAPK in the anti-cancer effects of DIM in human hepatocellular carcinoma cells.Methods: Cell proliferation was measured with a Cell Counting Kit-8 (CCK-8) and the clonogenic formation assay. Cell apoptosis was examined by flow cytometric analysis and Hoechst dye staining. Cleaved-caspase3, cleaved-PARP, Bax, total, and phosphorylated p38 MAPK were assayed by western blotting. [Ca2+]i was measured with Fluo-3/AM by fluorescence microscopy. A23187, a calcium ionophore, was used to increase [Ca2+]i levels.Results: DIM inhibited cell proliferation in both SMMC-7721 and HepG2 cells in a concentration- and time-dependent manner. DIM also enhanced phosphorylation of p38 MAPK (p-p38), which was attenuated by SB203580. The proliferation inhibition and apoptosis induction by DIM were also blunted. In addition, DIM increased [Ca2+]i in HCC cells, and this effect was inhibited by the calcium chelator, BAPTA-AM, resulting in reduced p-p38 MAPK activation and apoptosis in DIM-treated cells, though the proliferation inhibition by DIM was unchanged. However, the DIM-induced cell proliferation inhibition and apoptosis were significantly enhanced by A23187, a selective calcium ionophore, which was attributed to exaggerated p-p38 MAPK.Conclusions: The calcium ionophore enhanced DIM-induced anti-cancer effects in hepatocellular carcinoma cells, secondary to [Ca2+]i-dependent activation of p38 MAPK. Treatment with a combination of DIM and calcium ionophore may offer a new approach to enhance the chemotherapeutic efficacy in liver cancer. |
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| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-24T03:20:27Z |
| publishDate | 2019-10-01 |
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| spelling | doaj.art-a55d7beed8104626a7259bba13b857de2022-12-21T17:17:30ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122019-10-011010.3389/fphar.2019.01167461575Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPKYuanyue Jiang0Yuanyue Jiang1Yanfei Fang2Yanfei Fang3Yang Ye4Xinming Xu5Bingfang Wang6Jie Gu7Michael Aschner8Jian Chen9Rongzhu Lu10Rongzhu Lu11Department of Preventive Medicine and Public Health Laboratory Science, School of Medicine, Jiangsu University, Zhenjiang, ChinaDepartment of Pathology, Kunshan Hospital of Traditional Chinese Medicine, Suzhou, ChinaDepartment of Preventive Medicine and Public Health Laboratory Science, School of Medicine, Jiangsu University, Zhenjiang, ChinaDepartment of Gastroenterology, The First People's Hospital of Taicang City, Taicang Affiliated Hospital of Soochow University, Suzhou, ChinaDepartment of Preventive Medicine and Public Health Laboratory Science, School of Medicine, Jiangsu University, Zhenjiang, ChinaDepartment of General Surgery, Affiliated Kunshan Hospital, Jiangsu University School of Medicine, Suzhou, ChinaDepartment of Digestive Disease, Affiliated Kunshan Hospital, Jiangsu University School of Medicine, Suzhou, ChinaInstitute of Life Science, Jiangsu University, Zhenjiang, ChinaDepartment of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY, United StatesDepartment of General Surgery, Affiliated Kunshan Hospital, Jiangsu University School of Medicine, Suzhou, ChinaDepartment of Preventive Medicine and Public Health Laboratory Science, School of Medicine, Jiangsu University, Zhenjiang, ChinaCenter for Experimental Research, Affiliated Kunshan Hospital, Jiangsu University School of Medicine, Suzhou, ChinaPurpose: 3,3′-Diindolylmethane (DIM), derived from indole-3-carbinol (I3C) in the Brassica species of cruciferous vegetables, has anticancer effects, but its exact underlying mechanism of action is unknown. We explored the roles of cytosolic free calcium ([Ca2+]i) and p38 MAPK in the anti-cancer effects of DIM in human hepatocellular carcinoma cells.Methods: Cell proliferation was measured with a Cell Counting Kit-8 (CCK-8) and the clonogenic formation assay. Cell apoptosis was examined by flow cytometric analysis and Hoechst dye staining. Cleaved-caspase3, cleaved-PARP, Bax, total, and phosphorylated p38 MAPK were assayed by western blotting. [Ca2+]i was measured with Fluo-3/AM by fluorescence microscopy. A23187, a calcium ionophore, was used to increase [Ca2+]i levels.Results: DIM inhibited cell proliferation in both SMMC-7721 and HepG2 cells in a concentration- and time-dependent manner. DIM also enhanced phosphorylation of p38 MAPK (p-p38), which was attenuated by SB203580. The proliferation inhibition and apoptosis induction by DIM were also blunted. In addition, DIM increased [Ca2+]i in HCC cells, and this effect was inhibited by the calcium chelator, BAPTA-AM, resulting in reduced p-p38 MAPK activation and apoptosis in DIM-treated cells, though the proliferation inhibition by DIM was unchanged. However, the DIM-induced cell proliferation inhibition and apoptosis were significantly enhanced by A23187, a selective calcium ionophore, which was attributed to exaggerated p-p38 MAPK.Conclusions: The calcium ionophore enhanced DIM-induced anti-cancer effects in hepatocellular carcinoma cells, secondary to [Ca2+]i-dependent activation of p38 MAPK. Treatment with a combination of DIM and calcium ionophore may offer a new approach to enhance the chemotherapeutic efficacy in liver cancer.https://www.frontiersin.org/article/10.3389/fphar.2019.01167/full3,3′-diindolylmethanecytosolic Ca2+p38 MAPKhepatocellular carcinoma cellsapoptosisproliferation |
| spellingShingle | Yuanyue Jiang Yuanyue Jiang Yanfei Fang Yanfei Fang Yang Ye Xinming Xu Bingfang Wang Jie Gu Michael Aschner Jian Chen Rongzhu Lu Rongzhu Lu Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK Frontiers in Pharmacology 3,3′-diindolylmethane cytosolic Ca2+ p38 MAPK hepatocellular carcinoma cells apoptosis proliferation |
| title | Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK |
| title_full | Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK |
| title_fullStr | Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK |
| title_full_unstemmed | Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK |
| title_short | Anti-Cancer Effects of 3, 3’-Diindolylmethane on Human Hepatocellular Carcinoma Cells Is Enhanced by Calcium Ionophore: The Role of Cytosolic Ca2+ and p38 MAPK |
| title_sort | anti cancer effects of 3 3 diindolylmethane on human hepatocellular carcinoma cells is enhanced by calcium ionophore the role of cytosolic ca2 and p38 mapk |
| topic | 3,3′-diindolylmethane cytosolic Ca2+ p38 MAPK hepatocellular carcinoma cells apoptosis proliferation |
| url | https://www.frontiersin.org/article/10.3389/fphar.2019.01167/full |
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