title: Tackling the Cytotoxic Effect of a Marine Polycyclic Quinone-Type Metabolite: Halenaquinone Induces Molt 4 Cells Apoptosis via Oxidative Stress Combined with the Inhibition of HDAC and Topoisomerase Activities.
abstract: A marine polycyclic quinone-type metabolite, halenaquinone (HQ), was found to inhibit the proliferation of Molt 4, K562, MDA-MB-231 and DLD-1 cancer cell lines, with IC50 of 0.48, 0.18, 8.0 and 6.76 μg/mL, respectively. It exhibited the most potent activity against leukemia Molt 4 cells. Accumulating evidence showed that HQ may act as a potent protein kinase inhibitor in cancer therapy. To fully understand the mechanism of HQ, we further explored the precise molecular targets in leukemia Molt 4 cells. We found that the use of HQ increased apoptosis by 26.23%-70.27% and caused disruption of mitochondrial membrane potential (MMP) by 17.15%-53.25% in a dose-dependent manner, as demonstrated by Annexin-V/PI and JC-1 staining assays, respectively. Moreover, our findings indicated that the pretreatment of Molt 4 cells with N-acetyl-l-cysteine (NAC), a reactive oxygen species (ROS) scavenger, diminished MMP disruption and apoptosis induced by HQ, suggesting that ROS overproduction plays a crucial rule in the cytotoxic activity of HQ. The results of a cell-free system assay indicated that HQ could act as an HDAC and topoisomerase catalytic inhibitor through the inhibition of pan-HDAC and topoisomerase IIα expression, respectively. On the protein level, the expression of the anti-apoptotic proteins p-Akt, NFκB, HDAC and Bcl-2, as well as hexokinase II was inhibited by the use of HQ. On the other hand, the expression of the pro-apoptotic protein Bax, PARP cleavage, caspase activation and cytochrome c release were increased after HQ treatment. Taken together, our results suggested that the antileukemic effect of HQ is ROS-mediated mitochondrial apoptosis combined with the inhibitory effect on HDAC and topoisomerase activities.
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung 944, Taiwan. m6430005@hotmail.com.
authors.forename: Shou-Ping
authors.initials: SP
authors.lastname: Shih
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan. mg2253@yahoo.com.tw.|Division of Urology, Department of Surgery, Zuoying Branch of Kaohsiung Armed Forces General Hospital, Kaohsiung 813, Taiwan. mg2253@yahoo.com.tw.
authors.forename: Man-Gang
authors.initials: MG
authors.lastname: Lee
authors.affiliation: Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, Ain-Shams University, Organization of African Unity Street, Abassia, Cairo 11566, Egypt. elshazly444@googlemail.com.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El-Shazly
authors.affiliation: Department of Urology, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung 812, Taiwan. juanuro@gmail.com.|Department of Urology, College of Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan. juanuro@gmail.com.|Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. juanuro@gmail.com.
authors.forename: Yung-Shun
authors.initials: YS
authors.lastname: Juan
authors.affiliation: Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan. wzh@mail.nsysu.edu.tw.
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Wen
authors.affiliation: Department of Botanicals, Medical and Pharmaceutical Industry Technology and Development Center , New Taipei City 248, Taiwan. ycdu0626@gmail.com.
authors.forename: Ying-Chi
authors.initials: YC
authors.lastname: Du
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung 944, Taiwan. x2219@nmmba.gov.tw.|National Museum of Marine Biology & Aquarium, Pingtung 944, Taiwan. x2219@nmmba.gov.tw.
authors.forename: Jui-Hsin
authors.initials: JH
authors.lastname: Su
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung 944, Taiwan. pjsung@nmmba.gov.tw.|National Museum of Marine Biology & Aquarium, Pingtung 944, Taiwan. pjsung@nmmba.gov.tw.
authors.forename: Ping-Jyun
authors.initials: PJ
authors.lastname: Sung
authors.affiliation: The PhD Program of Cancer Biology and Drug discovery, China Medical University, Taichung 404, Taiwan. j520c@hotmail.com.
authors.forename: Yu-Cheng
authors.initials: YC
authors.lastname: Chen
authors.affiliation: School of Pharmacy, College of Pharmacy, China Medical University, Taichung 404, Taiwan. q9113054@yahoo.com.tw.|Chinese Medicine Research and Development Center, China Medical University Hospital, Taichung 404, Taiwan. q9113054@yahoo.com.tw.
authors.forename: Juan-Cheng
authors.initials: JC
authors.lastname: Yang
authors.affiliation: School of Pharmacy, College of Pharmacy, China Medical University, Taichung 404, Taiwan. yachwu@mail.cmu.edu.tw.|Chinese Medicine Research and Development Center, China Medical University Hospital, Taichung 404, Taiwan. yachwu@mail.cmu.edu.tw.|Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung 807, Taiwan. yachwu@mail.cmu.edu.tw.|Center of Molecular Medicine, China Medical University Hospital, Taichung 404, Taiwan. yachwu@mail.cmu.edu.tw.
authors.forename: Yang-Chang
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung 944, Taiwan. jinx6609@nmmba.gov.tw.|National Museum of Marine Biology & Aquarium, Pingtung 944, Taiwan. jinx6609@nmmba.gov.tw.
authors.forename: Mei-Chin
authors.initials: MC
authors.lastname: Lu
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm; D017209:Apoptosis / Q000187:drug effects*; D016227:Benzoquinones / Q000378:metabolism*; D045744:Cell Line, Tumor; D045304:Cytochromes c / Q000378:metabolism; D004250:DNA Topoisomerases, Type II; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors*; D005260:Female; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D020014:K562 Cells; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008819:Mice, Nude; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000951:Antigens, Neoplasm; D016227:Benzoquinones; D004268:DNA-Binding Proteins; D056572:Histone Deacetylase Inhibitors; D016328:NF-kappa B; D047428:Protein Kinase Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D011809:Quinones; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; C004532:quinone; C430084:halenaquinone; D045304:Cytochromes c; D011065:Poly(ADP-ribose) Polymerases; D006655:Histone Deacetylases; D004250:DNA Topoisomerases, Type II
keywords: halenaquinone; histone deacetylase (HDAC); mitochondria; reactive oxygen species (ROS); topoisomerase