title: Hinokitiol induces DNA damage and autophagy followed by cell cycle arrest and senescence in gefitinib-resistant lung adenocarcinoma cells.
abstract: Despite good initial responses, drug resistance and disease recurrence remain major issues for lung adenocarcinoma patients with epidermal growth factor receptor (EGFR) mutations taking EGFR-tyrosine kinase inhibitors (TKI). To discover new strategies to overcome this issue, we investigated 40 essential oils from plants indigenous to Taiwan as alternative treatments for a wide range of illnesses. Here, we found that hinokitiol, a natural monoterpenoid from the heartwood of Calocedrus formosana, exhibited potent anticancer effects. In this study, we demonstrated that hinokitiol inhibited the proliferation and colony formation ability of lung adenocarcinoma cells as well as the EGFR-TKI-resistant lines PC9-IR and H1975. Transcriptomic analysis and pathway prediction algorithms indicated that the main implicated pathways included DNA damage, autophagy, and cell cycle. Further investigations confirmed that in lung cancer cells, hinokitiol inhibited cell proliferation by inducing the p53-independent DNA damage response, autophagy (not apoptosis), S-phase cell cycle arrest, and senescence. Furthermore, hinokitiol inhibited the growth of xenograft tumors in association with DNA damage and autophagy but exhibited fewer effects on lung stromal fibroblasts. In summary, we demonstrated novel mechanisms by which hinokitiol, an essential oil extract, acted as a promising anticancer agent to overcome EGFR-TKI resistance in lung cancer cells via inducing DNA damage, autophagy, cell cycle arrest, and senescence in vitro and in vivo.
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan; Department of Laboratory, Kunming Branch, Taipei City Hospital, Taipei, Taiwan.
authors.forename: Lan-Hui
authors.initials: LH
authors.lastname: Li
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Ping
authors.initials: P
authors.lastname: Wu
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Jen-Yi
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Pei-Rong
authors.initials: PR
authors.lastname: Li
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Wan-Yu
authors.initials: WY
authors.lastname: Hsieh
authors.affiliation: Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University Medical College, Taipei, Taiwan.
authors.forename: Chao-Chi
authors.initials: CC
authors.lastname: Ho
authors.affiliation: Division of Wood Cellulose, Taiwan Forestry Research Institute, Taipei, Taiwan.
authors.forename: Chen-Lung
authors.initials: CL
authors.lastname: Ho
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan; Graduate Institute of Oncology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Wan-Jiun
authors.initials: WJ
authors.lastname: Chen
authors.affiliation: Division of Infectious Diseases, Kunming Branch, Taipei City Hospital, Taipei, Taiwan.
authors.forename: Chien-Chun
authors.initials: CC
authors.lastname: Wang
authors.affiliation: Division of Infectious Diseases, Kunming Branch, Taipei City Hospital, Taipei, Taiwan; Department of Medicine, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Muh-Yong
authors.initials: MY
authors.lastname: Yen
authors.affiliation: Department of Pathology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors.forename: Shun-Min
authors.initials: SM
authors.lastname: Yang
authors.affiliation: Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Huei-Wen
authors.initials: HW
authors.lastname: Chen
mesh_terms: D000165:Acridine Orange; D000230:Adenocarcinoma / Q000188:drug therapy*; D000077192:Adenocarcinoma of Lung; D017304:Annexin A5; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D001343:Autophagy / Q000187:drug effects*; D015153:Blotting, Western; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects; D029779:Cupressaceae / Q000737:chemistry*; D004249:DNA Damage / Q000187:drug effects*; D019008:Drug Resistance, Neoplasm; D066246:ErbB Receptors / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D000077156:Gefitinib; D020869:Gene Expression Profiling; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy*; D039821:Monoterpenes; D009822:Oils, Volatile / Q000032:analysis / Q000302:isolation & purification*; D011799:Quinazolines; D060888:Real-Time Polymerase Chain Reaction; D013624:Taiwan; D014334:Tropolone / Q000031:analogs & derivatives; D014410:Tumor Stem Cell Assay
chemical_list: D017304:Annexin A5; D000972:Antineoplastic Agents, Phytogenic; D039821:Monoterpenes; D009822:Oils, Volatile; D011799:Quinazolines; D014334:Tropolone; D066246:ErbB Receptors; D000165:Acridine Orange; D000077156:Gefitinib; C009479:beta-thujaplicin
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