This study investigated the role of N6-methyladenosine RNA methylation in liver regeneration following partial hepatectomy in mice.We created a liver-specific knockout mouse model by the deletion of Mettl3,a key compo...This study investigated the role of N6-methyladenosine RNA methylation in liver regeneration following partial hepatectomy in mice.We created a liver-specific knockout mouse model by the deletion of Mettl3,a key component of the N6-methyladenosine methyltransferase complex,using the albumin-Cre system.Mettl3 liver-specific knockout mice and their wild-type littermates were subjected to 2/3 partial hepatectomy.Transcriptomic changes in liver tissue at 48 h after partial hepatectomy were detected by RNA-seq.Immunohistochemistry and immunofluorescence were used to determine protein expression levels of Ki67,hepatocyte nuclear factor 4 alpha,and cytokeratin 19.Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling was also performed.Liver weight/body weight ratios after partial hepatectomy were significantly lower in Mettl3 liverspecific knockout mice than in wild-type mice at 48 h after 2/3 partial hepatectomy(3.1%±0.11%vs.2.7%±0.03%).Compared with wild-type littermates,Mettl3 liver-specific knockout mice showed reduced bromodeoxyuridine staining and reduced Ki-67 expression at 48 h after 2/3 partial hepatectomy.RNA-seq analysis showed that Mettl3 liver-specific knockout delayed the cell cycle progression in murine liver by downregulating the expression levels of genes encoding cyclins D1,A2,B1,and B2.Loss of Mettl3-mediated N6-methyladenosine function led to attenuated liver regeneration by altering the mRNA decay of suppressor of cytokine signaling 6,thereby inhibiting the phosphorylation of signal transducer and activator of transcription 3 during early liver regeneration.These results demonstrated the importance of N6-methyladenosine mRNA modification in liver regeneration and suggest that Mettl3 targeting might facilitate liver regeneration.展开更多
基金This work was supported by the National Key Research and Development Program of China(No.2017YFC0908100[Q.X.])the Key Clinical Subject Construction Project of Shanghai(No.shslczdzk05801).
文摘This study investigated the role of N6-methyladenosine RNA methylation in liver regeneration following partial hepatectomy in mice.We created a liver-specific knockout mouse model by the deletion of Mettl3,a key component of the N6-methyladenosine methyltransferase complex,using the albumin-Cre system.Mettl3 liver-specific knockout mice and their wild-type littermates were subjected to 2/3 partial hepatectomy.Transcriptomic changes in liver tissue at 48 h after partial hepatectomy were detected by RNA-seq.Immunohistochemistry and immunofluorescence were used to determine protein expression levels of Ki67,hepatocyte nuclear factor 4 alpha,and cytokeratin 19.Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling was also performed.Liver weight/body weight ratios after partial hepatectomy were significantly lower in Mettl3 liverspecific knockout mice than in wild-type mice at 48 h after 2/3 partial hepatectomy(3.1%±0.11%vs.2.7%±0.03%).Compared with wild-type littermates,Mettl3 liver-specific knockout mice showed reduced bromodeoxyuridine staining and reduced Ki-67 expression at 48 h after 2/3 partial hepatectomy.RNA-seq analysis showed that Mettl3 liver-specific knockout delayed the cell cycle progression in murine liver by downregulating the expression levels of genes encoding cyclins D1,A2,B1,and B2.Loss of Mettl3-mediated N6-methyladenosine function led to attenuated liver regeneration by altering the mRNA decay of suppressor of cytokine signaling 6,thereby inhibiting the phosphorylation of signal transducer and activator of transcription 3 during early liver regeneration.These results demonstrated the importance of N6-methyladenosine mRNA modification in liver regeneration and suggest that Mettl3 targeting might facilitate liver regeneration.
