Article Sidebar
Published:
May 25, 2021Keywords:
Article Details
Submission of a manuscript to this journal is a representation that the manuscript has not been published previously and is not under consideration for publication elsewhere.
All authors named in each manuscript would be required to sign a form (to be supplied by the Editor) so that they may retain their copyright in the article but to assign to us (the Publishers) and its licensees in perpetuity, in all forms, formats and media (whether known or created in the future) to (i) publish, reproduce, distribute, display and store the contribution, (ii) translate the contribution into other languages, create adaptations, reprints, include within collections and create summaries, extracts and/or abstracts of the contribution, (iii) create any other derivative works(s) based on the contribution, (iv) to exploit all subsidiary rights in the contribution, (v) the inclusion of electronic links from the contribution to third party material where-ever it may be located, and (vi) license any thrid party to do any or all of the above.
Main Article Content
DNA damage and necroptosis induced by peroxidase from proso millet in human colorectal cancer cells
Abstract
Purpose: To investigate the effects of cationic peroxidase from proso millet (PmPOD) on DNA damage and necroptosis in human colon cancer HCT116 and HT29 cells.
Methods: Cell necroptosis and cell cycle was stained using Annexin V-FITC and cell cycle kits, respectively, and evaluated by flow cytometry. Lipid raft on the membrane was disrupted by cholesterol depletion and the location of PmPOD observed by confocal microscopy. Comet assays were used to detect DNA damage, and different inhibitors were also used. Knockdown of p53 or ectopic p53 expression in HCT116 cells were transfected p53 siRNA and pCMV3-TP53-myc plasmid, and p53 expression analyzed by western blotting.
Results: Pre-treatment of HCT116 and HT29 cell lines with the specific necroptosis inhibitor Nec-1 prevented PmPOD-induced necroptosis, whereas the apoptosis inhibitor, z-VAD-fmk, had no effect. The entry of PmPOD is necessary for induction of DNA damage and necroptosis. Furthermore, PmPOD induced cell cycle arrest at S phase, as well as DNA DSBs in vivo, as reflected by numerous γ-H2AX foci in CRC cells. However, the tumor suppressor protein, p53, alleviated PmPOD-induced DNA damage and necroptosis.
Conclusions: These results demonstrate that PmPOD-induced DNA DSBs in CRC cells is the main cause of necroptosis, and that the tumor suppressor protein, p53, alleviates PmPOD-induced necroptosis by promoting p53-mediated repair pathways.
