Author:
Song Dongjian,Wang Qi,Yan Zechen,Su Meng,Zhang Hui,Shi Longyan,Fan Yingzhong,Zhang Qian,Yang Heying,Zhang Da,Liu Qiuliang
Abstract
Abstract
Background
The molecular mechanisms of osteosarcoma (OS) are complex. In this study, we focused on the functions of melanoma cell adhesion molecule (MCAM), methyltransferase 3 (METTL3) and insulin like growth factor 2 mRNA binding protein 1 (IGF2BP1) in OS development.
Methods
qRT-PCR assay and western blot assay were performed to determine mRNA and protein expression of MCAM, METTL3, IGF2BP1 and YY1. MTT assay and colony formation assay were conducted to assess cell proliferation. Cell apoptosis, invasion and migration were evaluated by flow cytometry analysis, transwell assay and wound-healing assay, respectively. Methylated RNA Immunoprecipitation (MeRIP), dual-luciferase reporter, Co-IP, RIP and ChIP assays were performed to analyze the relationships of MCAM, METTL3, IGF2BP1 and YY1. The functions of METTL3 and MCAM in tumor growth were explored through in vivo experiments.
Results
MCAM was upregulated in OS, and MCAM overexpression promoted OS cell growth, invasion and migration and inhibited apoptosis. METTL3 and IGF2BP1 were demonstrated to mediate the m6A methylation of MCAM. Functionally, METTL3 or IGF2BP1 silencing inhibited OS cell progression, while MCAM overexpression ameliorated the effects. Transcription factor YY1 promoted the transcription level of METTL3 and regulated METTL3 expression in OS cells. Additionally, METTL3 deficiency suppressed tumor growth in vivo, while MCAM overexpression abated the effect.
Conclusion
YY1/METTL3/IGF2BP1/MCAM axis aggravated OS development, which might provide novel therapy targets for OS.
Funder
National Natural Science Foundation of China
Henan Province Young and Middle-aged Health Science and Technology Innovation Talents Outstanding Youth Project
Provincial and Ministry Co-construction Project of Henan Provincial Medical Science and Technology Research Plan
Joint Co-construction Project of Henan Provincial Medical Science and Technology Research Plan
Key Scientific Research Projects of Higher Education Institutions in Henan Province
Publisher
Springer Science and Business Media LLC