In this study, it specifically shows that the novel m7G‐modified tRNA‐derived small RNA (tsRNA) m7G‐3′‐tiRNA LysTTT (mtiRL), catalyzed by the tRNA m7G‐modifying enzyme mettl1, specifically binds ANXA2, promoting its Tyr24 phosphorylation, by enhancing ANXA2–Yes1 interactions, leading to ANXA2 activation in bladder cancer (BC) cells and promotes BC malignancy. Here, ANXA2 is linked to breast cancer.