title: Bisindolylmaleimide alkaloid BMA-155Cl induces autophagy and apoptosis in human hepatocarcinoma HepG-2 cells through the NF-κB p65 pathway.
abstract: Bisindolylmaleimides, a series of derivatives of a PKC inhibitor staurosporine, exhibit potential as anti-cancer drugs and have received considerable attention in clinical trials. This study aims to investigate the effects of a bisindolylmaleimide alkaloid 155Cl (BMA-155Cl) with a novel structure on autophagy and apoptosis in human hepatocarcinoma HepG-2 cells in vitro and in vivo. The cell poliferation was assessed with a MTT assay. Autophagy was evaluated by MDC staining and TEM analysis. Apoptosis was investigated using Annexin V-FITC/PI and DAPI staining. The antitumor effects were further evaluated in nude mice bearing HepG-2 xenografts, which received BMA-155Cl (10, 20 mg/kg, ip) for 18 days. Autophagy- and apoptosis-associated proteins and their mRNA levels were examined with Western blotting, immunohistochemistry, and RT-PCR. BMA-155Cl (2.5-20 μmol/L) inhibited the growth of HepG-2 cells with IC50values of 16.62±1.34, 12.21±0.83, and 8.44±1.82 μmol/L at 24, 48, and 72 h, respectively. Furthermore, BMA-155Cl (5-20 μmol/L) dose-dependently induced autophagy and apoptosis in HepG-2 cells. The formation of autophagic vacuoles was induced by BMA-155Cl (10 μmol/L) at approximately 6 h and peaked at approximately 15 h. Pretreatment with 3-MA potentiated BMA-155Cl-mediated apoptotic cell death. This compound dose-dependently increased the mRNA and protein levels of Beclin-1, NF-κB p65, p53, and Bax, but decreased the expression of IκB and Bcl-2. Pretreatment with BAY 11-7082, a specific inhibitor of NF-κB p65, blocked BMA-155Cl-induced expression of autophagy- and apoptosis-associated proteins. BMA-155Cl administration effectively suppressed the growth of HepG-2 xenografts in vivo, and increased the protein expression levels of LC3B, Beclin-1, NF-κB p65, and Bax in vivo. We conclude that the NF-κB p65 pathway is involved in BMA-155Cl-triggered autophagy, followed by apoptosis in HepG-2 cells in vitro and in vivo. Hence, BMA-155Cl could be a promising pro-apoptotic candidate for developing as a novel anti-cancer drug.
authors.affiliation: School of Ocean, Shandong University, Weihai 264209, China.|School of Pharmaceutical Sciences, Shandong University, Ji-nan 250012, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Sun
authors.affiliation: School of Ocean, Shandong University, Weihai 264209, China.|School of Pharmaceutical Sciences, Shandong University, Ji-nan 250012, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao 266000, China.
authors.forename: Hong-Guang
authors.initials: HG
authors.lastname: Ma
authors.affiliation: School of Ocean, Shandong University, Weihai 264209, China.
authors.forename: Pu
authors.initials: P
authors.lastname: Sun
authors.affiliation: School of Ocean, Shandong University, Weihai 264209, China.
authors.forename: Qi-Lin
authors.initials: QL
authors.lastname: Wang
authors.affiliation: School of Ocean, Shandong University, Weihai 264209, China.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Zhang
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Ji-nan 250012, China.
authors.forename: Yue-Mao
authors.initials: YM
authors.lastname: Shen
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao 266000, China.
authors.forename: Wei-Ming
authors.initials: WM
authors.lastname: Zhu
authors.affiliation: School of Ocean, Shandong University, Weihai 264209, China.|School of Pharmaceutical Sciences, Shandong University, Ji-nan 250012, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D000071186:Beclin-1 / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D056945:Hep G2 Cells; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D026121:Indole Alkaloids / Q000494:pharmacology / Q000627:therapeutic use*; D007211:Indoles / Q000494:pharmacology / Q000627:therapeutic use*; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008301:Maleimides / Q000494:pharmacology / Q000627:therapeutic use*; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009368:Neoplasm Transplantation; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D015398:Signal Transduction; D051996:Transcription Factor RelA / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C000622032:BMA-155Cl; C494827:Bax protein, mouse; D000071186:Beclin-1; C115010:Becn1 protein, mouse; D020672:I-kappa B Proteins; D026121:Indole Alkaloids; D007211:Indoles; D008301:Maleimides; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051996:Transcription Factor RelA; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C052617:Bcl2 protein, mouse