Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling
Background: Dae-Hwang-Mok-Dan-Tang (DHMDT), a traditional Korean medicine, contains five species of medicinal plants and has been used to treat patients with digestive tract cancer for hundreds of years; however, its anticancer mechanism is poorly understood. In the present study, we investigated th...
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Elsevier
2017-06-01
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| Series: | Integrative Medicine Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2213422017300057 |
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| author | Cheol Park Su Hyun Hong Yung Hyun Choi |
| author_facet | Cheol Park Su Hyun Hong Yung Hyun Choi |
| author_sort | Cheol Park |
| collection | DOAJ |
| description | Background: Dae-Hwang-Mok-Dan-Tang (DHMDT), a traditional Korean medicine, contains five species of medicinal plants and has been used to treat patients with digestive tract cancer for hundreds of years; however, its anticancer mechanism is poorly understood. In the present study, we investigated the proapoptotic effects of DHMDT in human colon cancer HCT-116 cells. Methods: Cytotoxicity was evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Apoptosis was detected using 4,6-diamidino-2-phenyllindile staining, agarose gel electrophoresis, and flow cytometry. The protein levels were determined using Western blot analysis. Caspase activity was measured using a colorimetric assay. Results: Treatment with DHMDT resulted in a growth inhibition coupled with apoptosis induction, which was associated with the downregulation of members of IAP (inhibitor of apoptosis protein) family, including XIAP and survivin, and the activation of caspase-9 and -3 accompanied by proteolytic degradation of poly(ADP-ribose)-polymerase and phospholipase C-γ1. DHMDT treatment also showed a correlation with the translocation of proapoptotic Bax to mitochondria, the loss of mitochondrial membrane permeabilization, and the cytochrome c release from the mitochondria to the cytosol. Moreover, DHMDT increased the levels of death receptor-associated ligands and enhanced activation of caspase-8 and cleavage of its substrate, Bid. However, the pan-caspase inhibitor could reverse DHMDT-induced apoptosis. In addition, DHMDT suppressed the phosphoinositide 3-kinase (PI3K)/Akt pathway, and treatment with a potent inhibitor of PI3K further increased the apoptotic activity of DHMDT. Conclusion: Our data showed that DHMDT induces HCT-116 cell apoptosis by activating intrinsic and extrinsic apoptosis pathways and by suppressing the PI3K/Akt signal pathway; however, further studies are needed to identify the active compounds. Keywords: apoptosis, caspase, Dae-Hwang-Mok-Dan-Tang, HCT-116 cells, PI3K/Akt |
| format | Article |
| id | doaj-art-90ecaf7de7fa4b4984604199d312fe0d |
| institution | OA Journals |
| issn | 2213-4220 |
| language | English |
| publishDate | 2017-06-01 |
| publisher | Elsevier |
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| series | Integrative Medicine Research |
| spelling | doaj-art-90ecaf7de7fa4b4984604199d312fe0d2025-08-20T02:09:43ZengElsevierIntegrative Medicine Research2213-42202017-06-016217918910.1016/j.imr.2017.03.003Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signalingCheol Park0Su Hyun Hong1Yung Hyun Choi2Department of Molecular Biology, College of Natural Sciences, Dongeui University, Busan, KoreaDepartment of Biochemistry, Dongeui University College of Korean Medicine, Busan, KoreaDepartment of Biochemistry, Dongeui University College of Korean Medicine, Busan, Korea; Anti-Aging Research Center & Blue-Bio Industry Regional Innovation Center, Dongeui University, Busan, Korea; Corresponding author. Department of Biochemistry, Dongeui University College of Korean Medicine, 52-57 Yangjeong-ro, Busanjin-gu, Busan 47227, Republic of Korea.Background: Dae-Hwang-Mok-Dan-Tang (DHMDT), a traditional Korean medicine, contains five species of medicinal plants and has been used to treat patients with digestive tract cancer for hundreds of years; however, its anticancer mechanism is poorly understood. In the present study, we investigated the proapoptotic effects of DHMDT in human colon cancer HCT-116 cells. Methods: Cytotoxicity was evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Apoptosis was detected using 4,6-diamidino-2-phenyllindile staining, agarose gel electrophoresis, and flow cytometry. The protein levels were determined using Western blot analysis. Caspase activity was measured using a colorimetric assay. Results: Treatment with DHMDT resulted in a growth inhibition coupled with apoptosis induction, which was associated with the downregulation of members of IAP (inhibitor of apoptosis protein) family, including XIAP and survivin, and the activation of caspase-9 and -3 accompanied by proteolytic degradation of poly(ADP-ribose)-polymerase and phospholipase C-γ1. DHMDT treatment also showed a correlation with the translocation of proapoptotic Bax to mitochondria, the loss of mitochondrial membrane permeabilization, and the cytochrome c release from the mitochondria to the cytosol. Moreover, DHMDT increased the levels of death receptor-associated ligands and enhanced activation of caspase-8 and cleavage of its substrate, Bid. However, the pan-caspase inhibitor could reverse DHMDT-induced apoptosis. In addition, DHMDT suppressed the phosphoinositide 3-kinase (PI3K)/Akt pathway, and treatment with a potent inhibitor of PI3K further increased the apoptotic activity of DHMDT. Conclusion: Our data showed that DHMDT induces HCT-116 cell apoptosis by activating intrinsic and extrinsic apoptosis pathways and by suppressing the PI3K/Akt signal pathway; however, further studies are needed to identify the active compounds. Keywords: apoptosis, caspase, Dae-Hwang-Mok-Dan-Tang, HCT-116 cells, PI3K/Akthttp://www.sciencedirect.com/science/article/pii/S2213422017300057 |
| spellingShingle | Cheol Park Su Hyun Hong Yung Hyun Choi Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling Integrative Medicine Research |
| title | Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling |
| title_full | Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling |
| title_fullStr | Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling |
| title_full_unstemmed | Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling |
| title_short | Induction of apoptosis by Dae-Hwang-Mok-Dan-Tang in HCT-116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3-kinase/Akt signaling |
| title_sort | induction of apoptosis by dae hwang mok dan tang in hct 116 colon cancer cells through activation of caspases and inactivation of the phosphatidylinositol 3 kinase akt signaling |
| url | http://www.sciencedirect.com/science/article/pii/S2213422017300057 |
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