Establishment Of Transcriptome-wide M~6A Methylome Of Osteosarcoma Stem Cells Enriched By Chemotherapy

OBJECTIVES:To establish the transcriptome-wide N~6-methyladenosine methylome profile of osteosarcoma stem cells(OSCs)and further analyze the differences of m~6A methylation as well as gene expression between OSCs and paired parental cell lines.MATERIALS AND METHODS:Chemoresistant OSCs were enriched by doxorubicin treatment.The expression of m~6A-related enzymes was detected by q RT-PCR and western blot.Me RIP-seq and RNA-seq were performed to identify the differences in m~6A methylation and gene expression.Data analysis was conducted to explore the modified genes and their clinical significance.RESULTS:Three m~6A-related enzymes were altered in OSCs.Differentially methylated genes were enriched in some pathways regulating the pluripotency of stem cells.The expression of several candidate genes was found consistent with that in the GSE33458 dataset and associated with poor prognosis of osteosarcoma patients.CONCLUSIONS:m~6A methylation may play a significant role in acquired chemo-resistance of OS by activating signaling pathways regulating pluripotency of stem cells and inhibiting the osteoblast differentiation process so that OSCs might be enriched.FZD8,KLF4,BMP2,and SMAD5 might be candidate targets.