| Autor: |
Bhosale PB; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea., Abusaliya A; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea., Kim HH; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea., Ha SE; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea.; Biological Resources Research Group, Bioenvironmental Science & Toxicology Division, Korea Institute of Toxicology (KIT), 17 Jeigok-gil, Jinju 52834, Korea., Park MY; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea., Jeong SH; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea., Vetrivel P; Department of Pharmacy, National University of Singapore, Singapore 117643, Singapore., Heo JD; Department of Pharmacy, National University of Singapore, Singapore 117643, Singapore., Kim JA; Department of Physical Therapy, International University of Korea, Jinju 52833, Korea., Won CK; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea., Kim HW; Division of Animal Bioscience & Intergrated Biotechnology, Jinju 52725, Korea., Kim GS; Department of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea. |
| Abstrakt: |
Apigetrin (7-(β-D-glucopyranosyloxy)-4',5-dihydroxyflavone), a glycoside bioactive dietary flavonoid derived from Taraxacum officinale and Teucrium gnaphalodes, is known to possess anticancer, antioxidant, and anti-inflammatory effects on numerous cancers. In the present study, we examined the effect of apigetrin in Hep3B hepatocellular cancer cell line (HCC). Apigetrin inhibited cell growth and proliferation of Hep3B cells, as confirmed by MTT and colony formation assay. We used apigetrin at concentrations of 0, 50, and 100 µM for later experiments. Of these concentrations, 100 µM of apigetrin showed a significant effect on cell inhibition. In apigetrin-treated Hep3B cells, cell cycle arrest occurred at the G2/M phase. Apoptosis and necroptosis of Hep3B cells treated with apigetrin were confirmed by Annexin V/propidium iodide (PI) staining and flow cytometry results. Morphological observation through 4',6-diamidino-2-phenylindole (DAPI) staining showed intense blue fluorescence representing chromatin condensation. Hematoxylin staining showed necroptotic features such as formation of vacuoles and swelling of organelles. Apigetrin increased reactive oxygen species (ROS) levels in cells, based on fluorescence imaging. Furthermore, the underlying mechanism involved in the apoptosis and necroptosis was elucidated through western blotting. Apigetrin up-regulated TNFα, but down-regulated phosphorylation of p-p65, and IκB. Apigetrin inhibited the expression of Bcl-xl but increased Bax levels. Up-regulation of cleaved PARP and cleaved caspase 3 confirmed the induction of apoptosis in apigetrin-treated Hep3B cells. Additionally, necroptosis markers RIP3, p-RIP3, and p-MLKL were significantly elevated by apigetrin dose-dependently, suggesting necroptotic cell death. Taken together, our findings strongly imply that apigetrin can induce apoptosis and necroptosis of Hep3B hepatocellular cancer cells. Thus, apigetrin as a natural compound might have potential for treating liver cancer. |
