Synergistic Effect of Graphene Oxide Coated Nanotised Apigenin with Paclitaxel (GO-NA/PTX): A ROS Dependent Mitochondrial Mediated Apoptosis in Ovarian Cancer

Anticancer Agents Med Chem. 2017;17(12):1721-1732. doi: 10.2174/1871520617666170425094549.

Abstract

Background: Ovarian cancer is most lethal among all gynecologic malignancies. Paclitaxel (PTX) is well used chemotherapeutic regimen for cancer control; however its undesired toxicity has been a matter of concern for clinicians. Here, we used the graphene oxide coated nanotised apigenin (GO-NA) to enhance the efficacy of paclitaxel.

Objective: The combined use of paclitaxel (PTX) and nanotised apigenin (NA) may reduce the PTX dose and increase the efficacy.

Methods: GO and GO-Apigenin was prepared by modified Hummers method and the nanoparticles were characterized by dynamic light scattering and transmission electron microscopy. SKOV-3 cells were treated by DMSO, Group I (Control)-McCoy's 5A Medium, Group II-Paclitaxel (5nM), Group III- Nanotised Apigenin (GO-NA-10µM), Group IV- Paclitaxel (5nM) + GO-NA (10µM). Cell viability and IC-50 value were determined by MTT assay, synergism by Compusyn software, ROS by DCFH-DA assay, SOD activity by kit and MMP were examined by JC-1 and mitotracker/DAPI staining, cell cycle by flow cytometry, mRNA and protein level by Real Time-PCR and Western blot respectively Results: Results showed that GO-NA-PTX enhanced the anti-proliferative effect in synergistic manner as compare to GO-NA and PTX alone. GO-NA-PTX significantly suppressed the SOD activity, promotes the ROS accumulation, mitochondrial depolarization, DNA integrity and cell cycle arrest collectively accord the apoptosis. Results of immunocytochemistry, RT-PCR and western blot showed up-regulation of caspase-3, Bax, and down-regulation of Bcl-2.

Conclusion: The combination of PTX with GO-NA produces synergistic effects in SKOV-3 cells via the modulation of pro and anti-apoptotic gene and may reduce side effects of PTX.

Keywords: Apigenin nanoparticle; Bax; Bcl-2; SOD; apoptosis; caspase-3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / enzymology
  • Adenocarcinoma / metabolism
  • Adenocarcinoma / pathology*
  • Antineoplastic Agents, Phytogenic / administration & dosage
  • Antineoplastic Agents, Phytogenic / chemistry
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apigenin / administration & dosage
  • Apigenin / chemistry
  • Apigenin / pharmacology*
  • Apoptosis / drug effects*
  • Blotting, Western
  • Caspase 3 / genetics
  • Comet Assay
  • DNA Damage
  • Drug Synergism
  • Female
  • Flow Cytometry
  • Graphite / pharmacology*
  • Humans
  • Immunohistochemistry
  • Inhibitory Concentration 50
  • Membrane Potential, Mitochondrial / drug effects
  • Microscopy, Electron, Transmission
  • Nanoparticles / chemistry*
  • Ovarian Neoplasms / enzymology
  • Ovarian Neoplasms / metabolism
  • Ovarian Neoplasms / pathology*
  • Paclitaxel / chemistry*
  • Particle Size
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Scattering, Radiation
  • Superoxide Dismutase / metabolism
  • bcl-2-Associated X Protein / genetics

Substances

  • Antineoplastic Agents, Phytogenic
  • BAX protein, human
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • bcl-2-Associated X Protein
  • graphene oxide
  • Graphite
  • Apigenin
  • Superoxide Dismutase
  • CASP3 protein, human
  • Caspase 3
  • Paclitaxel