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Publication
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| Research Title |
13-cis-retinoic acid induces ROS-dependent apoptosis via a Bax-mediated intrinsic pathway in human cholangiocarcinoma cells |
| Date of Distribution |
15 February 2019 |
| Conference |
| Title of the Conference |
THE 41st PHARMACOLOGICAL AND THERAPEUTIC SOCIETY OF THAILAND MEETING |
| Organiser |
Department of Pharmacology, Faculty of Medicine, Chiang Mai University |
| Conference Place |
Chiangmai Grandview Hotel & Convention Center |
| Province/State |
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| Conference Date |
14 February 2019 |
| To |
15 February 2019 |
| Proceeding Paper |
| Volume |
41 |
| Issue |
41 |
| Page |
38 |
| Editors/edition/publisher |
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| Abstract |
INTRODUCTION: 13-cis-retinoic acid (13CRA), an active metabolite of vitamin A, plays a role in several physiological processes including embryonic development, cell differentiation, cell proliferation, cell death and survival. Recently, the anti-cancer activity of 13CRA has been reported; however, the effect of 13CRA in cholangiocarcinoma (CCA, an aggressive malignancy of bile duct epithelial) has not been shown.
OBJECTIVES: In this study, we aimed at investigating the effects of 13CRA in human CCA cell, KKU-100, as well as the detail molecular mechanism by which 13CRA induces apoptosis.
METHODS: Human CCA KKU-100 cells were used in this study. Apoptosis was analyzed by annexin V-PE and 7-AAD staining flow cytometry. Intracellular reactive oxygen species (ROS) generation was examined by dihydroethedium (DHE) method. Caspase-3 and -9 activations were measured by enzymatic assay. Expressions of proteins and mRNA were determined by western blot analysis and RT-qPCR, respectively.
RESULTS: Flow cytometry showed that 1.25 µM of 13CRA treatment for 48 hr effectively induced apoptosis cell death in human CCA KKU-100 cells. Fluorescent signal of ethidium demonstrated that 13CRA treatment for 90 min raised intracellular ROS production. Western blot analysis appeared that 13CRA treatment for 6 hr caused an increase of pro-apoptotic proteins; Bax, AIF, cytochrome C. 13CRA treatment for 12 hr significantly increased the activities of caspase-9 and -3, characteristics of the intrinsic apoptosis. Pretreatment with antioxidants, n-acetyl cysteine (NAC) or 4‐hydroxy‐2, 2, 6, 6‐tetramethylpiperidine‐1‐oxyl (TEMPOL) could completely block 13CRA-induced apoptosis. In addition, RT-qPCR after 13CRA treatment for 48 hr indicated down-regulation of Nrf-2, a master regulator of antioxidant system, and Nrf-2 targeted genes including NQO1, GCLC, and GSTP1.
CONCLUSIONS: Collectively, these results demonstrate that 13CRA induces a Bax-mediated intrinsic pathway in human CCA KKU-100 cells. The apoptosis effect of 13CRA is casually associated with increases oxidative stress partly through down-regulation of Nrf-2-targeted antioxidant genes. Thus, further studies highlighted the potential anti-cancer effect of 13CRA in CCA are warranted. |
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| Peer Review Status |
มีผู้ประเมินอิสระ |
| Level of Conference |
ชาติ |
| Type of Proceeding |
Abstract |
| Type of Presentation |
Poster |
| Part of thesis |
true |
| Presentation awarding |
false |
| Attach file |
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| Citation |
0
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Journal Publication
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