title: Mirabijalone E: a novel rotenoid from Mirabilis himalaica inhibited A549 cell growth in vitro and in vivo.
abstract: BACKGROUND
  The roots of Mirabilis himalaica have been used in Tibetan folk medicine for treatment of uterine cancer, nephritis edematous, renal calculus and arthrodynia. In our previous work, the ethanol extract of roots had shown potent cytotoxicity against human cancer cells. However, no information is available on the antitumor effect of Mirabilis himalaica. The aim of the present study was to investigate the active constituents guided by bioassay and evaluate the related antitumor efficacy in vitro and in vivo.
  
  
  METHODS
  The active subextract (ethyl acetate) was subjected to successive chemical separation using a combination of silica gel, LH-20 chromatography and semi-preparative HPLC. The structures were determined by spectroscopic analysis techniques such as nuclear magnetic resonance (NMR) and mass spectrometry. Three human cancer cell lines, A549, HepG2 and HeLa were used for in vitro cytotoxicity evaluation of all isolated compounds by MTT-assay. Then, the potent and novel compound mirabijalone E was employed to the mechanism study againstA549 cells. BrdU immunofluorescence, soft agar assay and cell cycle analysis were employed to detect the cell proliferation effects. Annexin V-FITC/PI staining assay was used for examining apoptotic effects. Expression levels of apoptosis-related proteins were determined by western blot assay. in vivo tumorigenic assay was used to evaluate the xenograft tumor growth treated with mirabijalone E.
  
  
  RESULTS
  One new rotenoid compound, mirabijalone E, together with eight known rotenoids was isolated from Mirabilis himalaica. Mirabijalone E, 9-O-methyl-inone B, boeravinone C and boeravinone H exhibited cytotoxicity against A 549 and HeLa cells. Further study on mirabijalone E was carried out in vitro and in vivo. Mirabijalone E inhibited A549 cells growth in a time and dose-dependent manner, which arrested cell cycle in S phase. Mechanistically, mirabijalone E treatment resulted in the increase of Bax expression level, the decrease of Bcl-2 level and the activation of caspase-3, which suggested the activation of apoptosis cascades. Consequently, the xenograft treated with mirabijalone E showed markedly suppressed tumor growth.
  
  
  CONCLUSIONS
  The result suggested that mirabijalone E, together with active compounds, 9-O-methyl-4-hydroxyboeravinone B, boeravinone C and boeravinone H could be a promising candidate for cancer therapy.
authors.affiliation: Key Laboratory on Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, PR China.
authors.forename: Lang
authors.initials: L
authors.lastname: Linghu
authors.affiliation: Key Laboratory on Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, PR China.
authors.forename: Haixia
authors.initials: H
authors.lastname: Fan
authors.affiliation: Key Laboratory on Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, PR China.
authors.forename: Yijie
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Key Laboratory on Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, PR China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Key Laboratory on Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, PR China.
authors.forename: Panpan
authors.initials: P
authors.lastname: Yang
authors.affiliation: Agriculture and Animal Husbandry College, Tibet University, Nyingchi, Tibet 860000, PR China.
authors.forename: Xiaozhong
authors.initials: X
authors.lastname: Lan
authors.affiliation: School of Life Sciences, Southwest University, Chongqing 400715, PR China.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Liao
authors.affiliation: Key Laboratory on Luminescence and Real-Time Analysis (Ministry of Education), College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, PR China. Electronic address: mminchen@swu.edu.cn.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D008297:Male; D038861:Medicine, Tibetan Traditional; D051379:Mice; D008818:Mice, Neurologic Mutants; D039282:Mirabilis / Q000737:chemistry*; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D018517:Plant Roots; D012402:Rotenone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000494:pharmacology; D059807:S Phase Cell Cycle Checkpoints / Q000187:drug effects; D013997:Time Factors; D023041:Xenograft Model Antitumor Assays
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D010936:Plant Extracts; C000596409:mirabijalone E; D012402:Rotenone
keywords: 9-O-methyl-4-hydroxyboeravinone B (PubChem CID: 487168); Apoptosis; Boeravinone A (PubChem CID: 14018346); Boeravinone B (PubChem CID: 14018348); Boeravinone C (PubChem CID: 13940642); Boeravinone D (PubChem CID: 15081178); Boeravinone E (PubChem CID: 11537197); Boeravinone F (PubChem CID: 12004175); Boeravinone H (PubChem CID: 16745324); Cell cycle; Cytotoxicity; Mirabijalone E; Mirabilis himalaica