Cytotoxicity Evaluation of Oxidized Single-Walled Carbon Nanotubes and Graphene Oxide on Human Hepatoma HepG2 cells: An iTRAQ-Coupled 2D LC-MS/MS Proteome Analysis

Abstract

Because of their attractive chemical and physical properties, graphitic nanomaterials and their derivatives have gained tremendous interest for applications in electronics, materials, and biomedical areas. However, few detailed studies have been performed to evaluate the potential cytotoxicity of these nanomaterials on living systems at the molecular level. In the present study, our group exploited the isobaric tagged relative and absolute quantification (iTRAQ)-coupled two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) approach with the purpose of characterizing the cellular functions in response to these nanomaterials at the proteome level. Specifically, the human hepatoma HepG2 cells were selected as the in vitro model to study the potential cytotoxicity of oxidized single-walled carbon nanotubes (SWCNTs) and graphene oxide (GO) on the vital organ of liver. Overall, 30 differentially expressed proteins involved in metabolic pathway, redox regulation, cytoskeleton formation, and cell growth were identified. Based on the protein profile, we found oxidized SWCNTs induced oxidative stress and interfered with intracellular metabolic routes, protein synthesis, and cytoskeletal systems. Further functional assays confirmed that oxidized SWCNTs triggered elevated level of reactive oxygen species (ROS), perturbed the cell cycle, and resulted in a significant increase in the proportion of apoptotic cells. However, only moderate variation of protein levels for the cells treated with GO was observed and functional assays further confirmed that GO was less cytotoxic in comparison to oxidized SWCNTs. These finding suggested that GO was more biocompatible and could be a promising candidate for bio-related applications.

Quercetin-mediated synthesis of graphene oxide–silver nanoparticle nanocomposites: a suitable alternative nanotherapy for neuroblastoma

“…81 Consistent with our results, previous studies also reported that GO can enter the cells by interacting with the plasma membrane and probably temporarily disrupting the phospholipid bilayer or causing plasma membrane invagination in HepG2 cells and neuroblastoma cell lines, including SK-N-BE(2) and SH-SY5Y. 30,112 Mari et al 30 further demonstrated that GO nanoribbons at low concentrations and within 72 h of exposure induced autophagy in neuroblastoma cell lines. Evidence from the presence of cytoplasmic vacuoles, especially in treated SK-N-BE(2) cells, and the induction of the lysosomal proteins LC3 and beclin, which are markers of autophagy, also confirmed that the autophagy is mainly due to ROS induced by GO.…”

Analysis of the cytotoxicity of hierarchical nanoporous graphenic carbon against human glioblastoma grade IV cells

“…25 Oxidative stress and generation of ROS can be involved in the toxic effects of graphene-based nanomaterials. 26 Assessment of ROS production showed a toxic influence of HNC on glioma cells. The interactions of the HNC particles with the U87 cells can lead to excessive ROS generation.…”

Single-Walled Carbon Nanohorn (SWNH) Aggregates Inhibited Proliferation of Human Liver Cell Lines and Promoted Apoptosis, Especially for Hepatoma Cell Lines

“…The liver cells were blocked in G2 phase by SWNHs, and the gap between DNA synthesis and mitosis was delayed by the quantity of SWNHs40 in HepG2, which may result from the apoptosis of HepG2 cells. 27 The cell cycle of liver cell lines was associated with the suppression of their proliferation induced by SWNHs.…”

“…34 Mitochondria, as the major source of reactive oxygen species (ROS) and redox machinery in cells, could activate inflammatory cytokines. 32 Oxidative stress had been found in HepG2 cells treated with unmodified or oxidized SWNTs and MWNTs, [25][26][27][28] L02 cells treated with unmodified, carboxyl-modified, or hydroxyl-modified MWNTs, 29 and rat hepatocytes treated with fullerenes C 60 , C 60 (OH) 12 , and C 60 (OH) 24 . 31 Gao et al 19 found that modified MWNTs internalized in lysosomes of human macrophages (THP-1 cells), generated ROS, and lowered mitochondrial membrane potential.…”