J Cancer 2020; 11(17):5007-5023. doi:10.7150/jca.46464 This issue
1. Department of Laboratory Medicine and Pathology, Stollery Children's Hospital, University of Alberta, Edmonton, Alberta, Canada.
2. Department of Laboratory Medicine and Pathology, Heritage Medical Research Centre, University of Alberta, Edmonton, Alberta, Canada.
3. Synthego Corporation, Menlo Park, CA 94025, USA.
4. Department of Paediatrics, University of Alberta, Edmonton, Alberta, Canada.
5. Department of Medical Genetics, University of Alberta, Edmonton, Alberta, Canada.
6. Department of Oncology, University of Alberta, Edmonton, Alberta, Canada.
7. National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan 430068, P.R. China.
8. Department of Orthopedics, Tianyou Hospital, Wuhan University of Science and Technology, Wuhan, Hubei, P.R. China.
Osteosarcoma affects both adolescents and adults, and some improvement in the survival rate for affected patients has been reached in the last decade. Still, non-specificity and systemic toxicity may limit traditional therapeutic approaches to some extent. The insulin growth factor 1 (IGF1) and its binding protein (IGFBP3) have been implicated in the tumorigenesis. Nanoparticles, such as graphene oxide (GO), can provide an effective treatment for cancer as they can specifically target cancer cells while reducing undesired side effects. This study aimed to evaluate the toxicity of GO on osteosarcoma in vitro using tumor cell lines with and without knocking out the IGF and IGFBP3 genes. Human osteosarcoma cell lines, U2OS and SAOS2, and the normal osteoblast cell line hFOB1.19 were used. The IGF1 and IGFBP3 genes were eliminated using CRISPR/Cas9. Tumor cells were cultured and treated with GO. Apoptosis and reactive oxygen species (ROS) were analyzed by Annexin V-FITC and ROS assays. The nuclear factor erythroid 2-related factor 2 (NRF2), which is a crucial regulator of cellular resistance to oxidants, was investigated by Western blotting. We found a significantly higher rate of apoptosis in the OS than hFOB1.19, especially in U2OS cells in which IGF1 and IGFBP3 were knocked out. ROS increase due to GO exposure was remarkably time and concentration-dependent. Based on the rate of apoptosis, ROS, Nrf-2 decrease, and cytomorphological changes, GO has a significant cytotoxic effect against OS. Targeting the IGF1 and IGFBP3 signaling pathway may strengthen GO-related cytotoxicity with the potential to increase the survival of patients affected by this tumor.
Keywords: Osteosarcoma, cell line, apoptosis, ROS, CRISPR-Cas9, IGF1, IGFBP3, Graphene oxide