• Cx58 is upregulated in NSCLC and its expression correlates with metastatic potential.
• MEF2B transcriptionally activates Cx58 expression, forming the MEF2B/Cx58 axis.
• Knockdown of Cx58 or MEF2B reduces NSCLC cell migration and invasion via cytoskeleton rearrangement.
• Targeting the MEF2B/Cx58 axis represents a potential therapeutic strategy for metastatic NSCLC.