Chemical composition and cytotoxic properties of Clinacanthus nutans root extracts

Pharm Biol. 2017 Dec;55(1):394-401. doi: 10.1080/13880209.2016.1242145.

Abstract

Context: 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.

Materials and 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 IC50 values 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.

Discussion and 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.

Keywords: BAX; BCL2; Clinacanthus nutans; anti-proliferative; apoptosis.

MeSH terms

  • Acanthaceae / chemistry*
  • Animals
  • Antineoplastic Agents, Phytogenic / isolation & purification
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apoptosis / drug effects*
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • Cell Nucleus Shape / drug effects
  • Cell Proliferation / drug effects*
  • Chromatin Assembly and Disassembly / drug effects
  • Dose-Response Relationship, Drug
  • Female
  • Flow Cytometry
  • Gas Chromatography-Mass Spectrometry
  • HeLa Cells
  • Humans
  • Inhibitory Concentration 50
  • MCF-7 Cells
  • Membrane Potential, Mitochondrial / drug effects
  • Mice
  • NIH 3T3 Cells
  • Phytotherapy
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Plant Roots / chemistry*
  • Plants, Medicinal
  • Reverse Transcriptase Polymerase Chain Reaction
  • Solvents / chemistry
  • Uterine Cervical Neoplasms / drug therapy*
  • Uterine Cervical Neoplasms / genetics
  • Uterine Cervical Neoplasms / metabolism
  • Uterine Cervical Neoplasms / pathology

Substances

  • Antineoplastic Agents, Phytogenic
  • Apoptosis Regulatory Proteins
  • Plant Extracts
  • Solvents