Abstract |
The objective of the present study was to evaluate the tumor and apoptotic effects of dihydromethysticin kavalactone against human osteosarcoma (MG-63) cells. Antiproliferative activity was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis induction by dihydromethysticin was demonstrated by fluorescence microscopy, quantitative videomicroscopy and Annexin V-FITC apoptosis detection kit. Mitochondrial membrane potential disruption was demonstrated by rhodamine-123 dye using flow cytometry. We also evaluated the effect of dihydromethysticin on PI3K/Akt pathway with an immunoblotting analysis. The results showed that the compound induced dose-dependent as well as time-dependent antiproliferative effects against MG-63 cell growth. Cell death and apoptotic body formation was noticed followed dihydromethysticin treatment at various doses. The percentage of apoptotic cells (early apoptosis+late apoptosis) increased from 6.63% in untreated control to 23.92%, 23.81% and 93.9% in 25 µM, 75 µM and 100 µ Mdihydromethysticin-treated cells respectively. Flow cytometric analysis showed dihydromethysticin induced an increase in G0/G1 cells (apoptotic cells). Furthermore, we observed mitochondrial transmembrane depolarization along with decreased phosphorylation levels for PI3K, AKT (Ser 473), AKT (Thr 308), GSK-3β, and BAD. These reductions were associated with down regulation of AKT and upregulation of both GSK-3β and BAD.
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Authors | Jun-Qi Dai, Yi-Gang Huang, Ai-Na He |
Journal | International journal of clinical and experimental pathology
(Int J Clin Exp Pathol)
Vol. 8
Issue 5
Pg. 4356-66
( 2015)
ISSN: 1936-2625 [Electronic] United States |
PMID | 26191127
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- 7,8-dihydromethysticin
- Antineoplastic Agents
- Pyrones
- Phosphatidylinositol 3-Kinases
- Proto-Oncogene Proteins c-akt
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Topics |
- Antineoplastic Agents
(pharmacology)
- Apoptosis
(drug effects)
- Blotting, Western
- Bone Neoplasms
(pathology)
- Cell Cycle Checkpoints
(drug effects)
- Cell Line, Tumor
- Cell Proliferation
(drug effects)
- Cell Survival
(drug effects)
- Flow Cytometry
- Humans
- Membrane Potential, Mitochondrial
(drug effects)
- Microscopy, Fluorescence
- Osteosarcoma
(pathology)
- Phosphatidylinositol 3-Kinases
(metabolism)
- Proto-Oncogene Proteins c-akt
(metabolism)
- Pyrones
(pharmacology)
- Signal Transduction
(drug effects, physiology)
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