The N6-methyladenosine pattern of MAP3K7 mediates the effects of sevoflurane on macrophage M2 polarization and cervical cancer migration and invasion

Introduction: The study was designed to determine whether and how sevoflurane (Sev) regulates tumor associated macrophage (TAM) polarization and cervical cancer (CC) cell progression. Material and methods: The M2 polarized THP-1 was treated with 3%Sev. The culture supernatant of M2 polarized THP-1 was co-cultured with the CC cell line Hela. The NF-kappa B activity was determined by luciferase reporter assay. The key genes dis-regulated by 3%Sev were determined by RNA sequencing (RNA-seq) followed by real-time reverse transcription PCR (qRT-PCR) assay. Luciferase reporter assay was used to analyze the function of 3%Sev based on N6-methyladenosine (m6A) site activity on MAP3K7 3 ' untranslated regions (3 ' UTRs). RNA immunoprecipitation (IP) using an anti-m6A antibody (anti-m6A RNA-IP) was performed to determine the m6A levels at MAP3K7 3 ' UTR. Results: 3%Sev treatment significantly up-regulated the M2 polarization markers and down-regulated the NF-kappa B activity of THP-1. Meanwhile, 3%Sev treated macrophages could enhance the migratory and invasive potential of CC cells. Further, 3%Sev significantly regulated the NF-kappa B pathway, including MAP3K7 inhibition. MAP3K7 overexpression reversed the 3%Sev-regulated NF-kappa B activity and M2 polarization. 3%Sev treatment increased m6A levels in the 3 ' UTR of MAP3K7. Mutational analysis of potential m6A sites within MAP3K7 3 ' UTR revealed that these sites were required for 3%Sev regulation. In conclusion, the m6A pattern of MAP3K7 mediates the effects of 3%Sev on macrophage M2 polarization and cervical cancer progression. Conclusions: 3%Sev enhanced TAMs M2 polarization through regulating the m6A pattern of MAP3K7, and therefore enhanced the stimulatory effect of M2 TAMs on the migration and invasion of CC cells.