Study On The Molecular Mechanism Of MKRN2 In Nervous System Tumors

MKRN2 is an E3 ubiquitin ligase.The biological role of MKRN2 in nervous system tumors is unclear.The purpose of this study is to explore the molecular mechanism of MKRN2 in regulating nervous system tumors.In this paper,we designed sh RNA targeting MKRN2 and established a stable cell line that knocked down MKRN2 in human neuroblastoma SHSY5 Y cells.Cell proliferation experiments,cell clone formation,and scratch test results show that knocking down MKRN2 can promote the proliferation and migration of SHSY5 Y cells.A series of experiments were carried out in this paper: protein interaction mass spectrometry,immunoprecipitation,GSTPulldown,and immunofluorescence.IGF2BP3 was identified as a new interacting protein of MKRN2,and through in vivo ubiquitination,endogenous ubiquitination,in The reconstructed ubiquitination system and mass spectrometry system in bacteria analyzed post-translational modification sites,etc.,and identified IGF2BP3 as a new ubiquitination substrate of MKRN2.IGF2BP3 is an RNA-binding protein that can regulate the stability of many m RNAs including PDPN,CD44,and CD164.Real-time fluorescence quantitative PCR and luciferase reporter gene experiments show that MKRN2 regulates CD44,PDPN,and IGF2BP3-dependent manner.CD164 expression.These results suggest that MKRN2 may be a potential target for the treatment of nervous system tumors.