Suppressing growth, migration, and invasion of human hepatocellular carcinoma HepG2 cells by Catharanthus roseus‑silver nanoparticles

• Published in: Toxicology in vitro 2020-09, Vol.67, p.104910, Article 104910
• Main Authors: Azhar, Nur Asna, Ghozali, Siti Zulaikha, Abu Bakar, Siti Aishah, Lim, Vuanghao, Ahmad, Nor Hazwani
• Format: Article
• Language: English
• Subjects: Antineoplastic Agents - administration & dosage; Antineoplastic Agents - chemistry; Apoptosis; Apoptosis - drug effects; Bioassays; Biosynthesis; Carcinoma, Hepatocellular - drug therapy; Catharanthus; Catharanthus roseus; Cell cycle; Cell Cycle Checkpoints - drug effects; Cell death; Cell Movement - drug effects; Cell proliferation; Cell Proliferation - drug effects; Comet assay; Cytotoxicity; Damage assessment; Damage detection; Deoxyribonucleic acid; DNA; DNA Damage; Endosomes; Evaluation; Hep G2 Cells; Hepatocellular carcinoma; Hepatocytes; HepG2; Humans; Iodides; Liver cancer; Liver Neoplasms - drug therapy; Membrane potential; Membrane Potential, Mitochondrial - drug effects; Metal Nanoparticles - administration & dosage; Metal Nanoparticles - chemistry; Mitochondria; Mortality; Nanoparticles; Nitric oxide; Nitric Oxide - metabolism; Plant Extracts - chemistry; Plant Leaves; Propidium iodide; Reactive oxygen species; Reactive Oxygen Species - metabolism; Silver; Silver - administration & dosage; Silver - chemistry; Silver nanoparticles; Silver Nitrate - chemistry; Time dependence; Toxicity; Transmission electron microscopy
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2020.104910

Application of silver nanoparticles serves as a new approach in cancer treatment due to its unique features. Biosynthesis of silver nanoparticles using plant is advantageous since they are easily accessible, nontoxic and produce quicker reaction compared to other methods. To evaluate the cytotoxicity, mechanism of cell death and DNA damage of biosynthesized Catharanthus roseus-silver nanoparticles on human liver cancer (HepG2) cells. The antiproliferative activity of Catharanthus roseus‑silver nanoparticles was measured using MTT assay. The cytotoxic effects were further evaluated by measuring nitric oxide and reactive oxygen species (ROS). The mechanism of cell death was determined by annexin-FITC/propidium iodide, mitochondrial membrane potential (MMP) and cell cycle assays. The assessment of DNA damage was evaluated using Comet assay method. The uptake of the nanoparticles were evaluated by Transmission Electron Microscopy (TEM). Catharanthus roseus‑silver nanoparticles has inhibited the proliferation of HepG2 cells in a time-dependent manner with a median IC50 value of 3.871 ± 0.18 μg/mL. The concentration of nitrite and ROS were significantly higher than control. The cell death was due to apoptosis associated with MMP loss, cell cycle arrest, and extensive DNA damage. TEM analysis indicated the presence of free nanoparticles and endosomes containing the nanoparticles. The findings show that Catharanthus roseus‑silver nanoparticles have produced cytotoxic effects on HepG2 cells and thus may have a potential to be used as an anticancer treatment, particularly for hepatocellular carcinoma. •Inhibition of cell proliferation was in parallel with apoptosis in time-dependent manner•Production of ROS and NO triggered MMP depolarization and DNA damage-induced cell cycle arrest•Intracellular endosomes indicating deposition of nanoparticles were mostly located adjacent to the cell membrane•C. roseus-silver nanoparticles may serve as a new approach for liver cancer treatment due to its unique features