title: Chemical composition and cytotoxic properties of Clinacanthus nutans root extracts.
abstract: BACKGROUND
  Clinacanthus nutans Lindau (Acanthaceae) is a medicinal plant that has been reported to have anti-inflammatory, antiviral, antimicrobial and antivenom activities. In Malaysia, it has been widely claimed to be effective in various cancer treatments but scientific evidence is lacking.
  
  
  OBJECTIVE
  This study investigates the chemical constituents, anti-proliferative, and apoptotic properties of C. nutans root extracts.
  
  
  METHODS
  The roots were subjected to solvent extraction using methanol and ethyl acetate. The anti-proliferative effects of root extracts were tested at the concentrations of 10 to 50 μg/mL on MCF-7 and HeLa by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay for 72 h. Morphological changes were observed under light microscope. Pro-apoptotic effects of root extracts were examined using flow cytometric analysis and RT-PCR. The chemical compositions of root extracts were detected using GC-MS.
  
  
  RESULTS
  The proliferation of MCF-7 cells was inhibited with the IC50values of 35 and 30 μg/mL, respectively, for methanol and ethyl acetate root extracts. The average inhibition of HeLa cells was ∼25%. Induction of apoptosis in MCF-7 was supported by chromatin condensation, down-regulation of BCL2 and unaltered expression of BAX. However, only ethyl acetate extract caused the loss of mitochondrial membrane potential. GC-MS analysis revealed the roots extracts were rich with terpenoids and phytosterols.
  
  
  CONCLUSIONS
  The results demonstrated that root extracts promote apoptosis by suppressing BCL2 via mitochondria-dependent or independent manner. The identified compounds might work solely or cooperatively in regulating apoptosis. However, further studies are required to address this.
authors.affiliation: a Biotechnology Research Institute, Universiti Malaysia Sabah , Jalan UMS , Kota Kinabalu , Sabah , Malaysia.
authors.forename: Peik Lin
authors.initials: PL
authors.lastname: Teoh
authors.affiliation: a Biotechnology Research Institute, Universiti Malaysia Sabah , Jalan UMS , Kota Kinabalu , Sabah , Malaysia.
authors.forename: Angelina Ying Fang
authors.initials: AY
authors.lastname: Cheng
authors.forename: Monica
authors.initials: M
authors.lastname: Liau
authors.affiliation: a Biotechnology Research Institute, Universiti Malaysia Sabah , Jalan UMS , Kota Kinabalu , Sabah , Malaysia.
authors.forename: Fui Fui
authors.initials: FF
authors.lastname: Lem
authors.affiliation: a Biotechnology Research Institute, Universiti Malaysia Sabah , Jalan UMS , Kota Kinabalu , Sabah , Malaysia.
authors.forename: Grace P
authors.initials: GP
authors.lastname: Kaling
authors.affiliation: b Faculty of Science & Natural Resources, Universiti Malaysia Sabah , Jalan UMS, Kota Kinabalu , Sabah , Malaysia.
authors.forename: Fern Nie
authors.initials: FN
authors.lastname: Chua
authors.affiliation: a Biotechnology Research Institute, Universiti Malaysia Sabah , Jalan UMS , Kota Kinabalu , Sabah , Malaysia.
authors.forename: Bo Eng
authors.initials: BE
authors.lastname: Cheong
mesh_terms: D027901:Acanthaceae / Q000737:chemistry*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D053144:Cell Nucleus Shape / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D005434:Flow Cytometry; D008401:Gas Chromatography-Mass Spectrometry; D006367:HeLa Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D061986:MCF-7 Cells; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D041681:NIH 3T3 Cells; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D018517:Plant Roots / Q000737:chemistry*; D010946:Plants, Medicinal; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012997:Solvents / Q000737:chemistry; D002583:Uterine Cervical Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; D010936:Plant Extracts; D012997:Solvents
keywords: BAX; BCL2; Clinacanthus nutans; anti-proliferative; apoptosis