Fig. 5
From: METTL3 drives NSCLC metastasis by enhancing CYP19A1 translation and oestrogen synthesis
METTL3 mediates m6A modification of CYP19A1 and promotes the synthesis of E2. (A) WB experiments demonstrated that knockdown of METTL3 using siRNA significantly reduced the protein expression of CYP19A1. (B) PCR experiments showed an upwards trend in the mRNA expression of CYP19A1 following siRNA knockdown of METTL3. (C) ELISA revealed a decrease in intracellular and extracellular levels of E2 after siRNA knockdown of METTL3. (D) CYP19A1 is upregulated after transfection of plasmids for METTL3 overexpression. (E) Protein expression of CYP19A1 is significantly downregulated after CRISPR‒Cas9 knockout of METTL3. (F) PCR indicated an upwards trend in the mRNA expression of CYP19A1 following CRISPR‒Cas9 knockout of METTL3. (G) ELISA demonstrated a significant decrease in intracellular and extracellular levels of E2 after CRISPR‒Cas9 knockout of METTL3. (H) Treatment with 5 µM STM2457 for 3 days led to a significant decrease in the protein level of CYP19A1. (I) Treatment with 5 µM STM2457 for 3 days led to a significant increase in the mRNA level of CYP19A1. (J) Treatment with 5 µM STM2457 for 3 days led to a significant reduction in E2 levels. (K) MeRIP-qPCR revealed that CYP19A1 mRNA was enriched in m6A modification and the relative levels of m6A whin CYP19A1 were significantly downregulated after METTL3 knockout in A549 cells. (L) MeRIP-qPCR revealed that CYP19A1 mRNA was enriched in m6A modification in H1975 cells. (M-N) CYP19A1 protein expression and intratumoural E2 concentration in lung metastasis of KO-METTL3 tumours are significantly downregulated