PMID- 33573023 OWN - NLM STAT- MEDLINE DCOM- 20210907 LR - 20240330 IS - 1422-0067 (Electronic) IS - 1422-0067 (Linking) VI - 22 IP - 3 DP - 2021 Jan 29 TI - Artemisia annua L. Polyphenol-Induced Cell Death Is ROS-Independently Enhanced by Inhibition of JNK in HCT116 Colorectal Cancer Cells. LID - 10.3390/ijms22031366 [doi] LID - 1366 AB - c-Jun N-terminal kinase (JNK) is activated by chemotherapeutic reagents including natural plant polyphenols, and cell fate is determined by activated phospho-JNK as survival or death depending on stimuli and cell types. The purpose of this study was to elucidate the role of JNK on the anticancer effects of the Korean plant Artemisia annua L. (pKAL) polyphenols in p53 wild-type HCT116 human colorectal cancer cells. Cell morphology, protein expression levels, apoptosis/necrosis, reactive oxygen species (ROS), acidic vesicles, and granularity/DNA content were analyzed by phase-contrast microscopy; Western blot; and flow cytometry of annexin V/propidium iodide (PI)-, dichlorofluorescein (DCF)-, acridine orange (AO)-, and side scatter pulse height (SSC-H)/DNA content (PI)-stained cells. The results showed that pKAL induced morphological changes and necrosis or late apoptosis, which were associated with loss of plasma membrane/Golgi integrity, increased acidic vesicles and intracellular granularity, and decreased DNA content through downregulation of protein kinase B (Akt)/beta-catenin/cyclophilin A/Golgi matrix protein 130 (GM130) and upregulation of phosphorylation of H2AX at Ser-139 (gamma-H2AX)/p53/p21/Bak cleavage/phospho-JNK/p62/microtubule-associated protein 1 light chain 3B (LC3B)-I. Moreover, JNK inhibition by SP600125 enhanced ROS-independently pKAL-induced cell death through downregulation of p62 and upregulation of p53/p21/Bak cleavage despite a reduced state of DNA damage marker gamma-H2AX. These findings indicate that phospho-JNK activated by pKAL inhibits p53-dependent cell death signaling and enhances DNA damage signaling, but cell fate is determined by phospho-JNK as survival rather than death in p53 wild-type HCT116 cells. FAU - Jung, Eun Joo AU - Jung EJ AD - Department of Biochemistry, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 52727, Korea. AD - Department of Internal Medicine, Institute of Health Sciences, Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju 52727, Korea. FAU - Paramanantham, Anjugam AU - Paramanantham A AD - Department of Internal Medicine, Institute of Health Sciences, Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju 52727, Korea. AD - Research Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea. FAU - Kim, Hye Jung AU - Kim HJ AUID- ORCID: 0000-0002-7067-6810 AD - Department of Pharmacology, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 52727, Korea. FAU - Shin, Sung Chul AU - Shin SC AD - Department of Chemistry, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea. FAU - Kim, Gon Sup AU - Kim GS AUID- ORCID: 0000-0002-0844-1009 AD - Research Institute of Life Science, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea. FAU - Jung, Jin-Myung AU - Jung JM AD - Department of Neurosurgery, Institute of Health Sciences, Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju 52727, Korea. FAU - Ryu, Chung Ho AU - Ryu CH AD - Department of Food Technology, Research Institute of Life Science, Gyeongsang National University, Jinju 52828, Korea. FAU - Hong, Soon Chan AU - Hong SC AD - Department of Surgery, Institute of Health Sciences, Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju 52727, Korea. FAU - Chung, Ky Hyun AU - Chung KH AD - Department of Urology, Institute of Health Sciences, Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju 52727, Korea. FAU - Kim, Choong Won AU - Kim CW AD - Department of Biochemistry, Institute of Health Sciences, Gyeongsang National University School of Medicine, Jinju 52727, Korea. FAU - Lee, Won Sup AU - Lee WS AUID- ORCID: 0000-0003-0863-1424 AD - Department of Internal Medicine, Institute of Health Sciences, Gyeongsang National University Hospital, Gyeongsang National University School of Medicine, Jinju 52727, Korea. LA - eng GR - 2017R1D1A3B05030971/Basic Science Research Program through the National Research Foundation of Korea (NRF)/ PT - Journal Article DEP - 20210129 PL - Switzerland TA - Int J Mol Sci JT - International journal of molecular sciences JID - 101092791 RN - 0 (Antineoplastic Agents, Phytogenic) RN - 0 (Polyphenols) RN - 0 (Protein Kinase Inhibitors) RN - 0 (Reactive Oxygen Species) RN - EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases) SB - IM MH - Antineoplastic Agents, Phytogenic/chemistry/*pharmacology MH - *Artemisia annua/chemistry MH - Cell Death/drug effects MH - Colorectal Neoplasms/*drug therapy/metabolism MH - HCT116 Cells MH - Humans MH - JNK Mitogen-Activated Protein Kinases/*antagonists & inhibitors/metabolism MH - Polyphenols/chemistry/*pharmacology MH - Protein Kinase Inhibitors/chemistry/*pharmacology MH - Reactive Oxygen Species/metabolism PMC - PMC7866371 OTO - NOTNLM OT - Artemisia annua L. OT - JNK OT - ROS OT - SP600125 OT - cell death OT - colorectal cancer OT - p53 OT - polyphenols COIS- The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. EDAT- 2021/02/13 06:00 MHDA- 2021/09/08 06:00 PMCR- 2021/01/29 CRDT- 2021/02/12 01:03 PHST- 2020/12/31 00:00 [received] PHST- 2021/01/25 00:00 [revised] PHST- 2021/01/26 00:00 [accepted] PHST- 2021/02/12 01:03 [entrez] PHST- 2021/02/13 06:00 [pubmed] PHST- 2021/09/08 06:00 [medline] PHST- 2021/01/29 00:00 [pmc-release] AID - ijms22031366 [pii] AID - ijms-22-01366 [pii] AID - 10.3390/ijms22031366 [doi] PST - epublish SO - Int J Mol Sci. 2021 Jan 29;22(3):1366. doi: 10.3390/ijms22031366.