Study on ubiquitination modification signal pathway in spermatogonia and its relationship with reproductive ability
Objective To explore the molecular mechanisms underlying the expression and function of ring finger protein 216(RNF216)in spermatogonia.Methods siRNA targeting RNF216 and pcDNA3.1-Flag-RNF216(RNF216 overexpression plasmid)were transfected into mouse spermatogonial cell line(GC-1 cell)to knock down or up-regulate RNF216 expression in cells.Cell proliferation was determined by cell counting kit 8(CCK-8)and crystal violet cell proliferation test,and cell migration ability was determined by Transwell test.The expression of RNF216 and toll-like receptor 4(TLR4)was detected by protein blot.Results Compared with si-NC group,the number of cell clones and migration in si-RNF216#l group and si-RNF216#2 group decreased significantly(P＜0.001).Compared with NC group,the number of clones and migration of GC-1 cells in RNF216 group increased significantly(P＜0.001).In cells treated with proteasome inhibitor MG132,the overexpression of RNF216 significantly increased the ubiquitination of TLR4(P＜0.05).Compared with RNF216 group,RNF216+TLR4 group GC-1 cells significantly decreased in cell viability,clone number and migration number(P＜0.05).Conclusions RNF216 can promote the migration and proliferation of GC-1 cells by degrading TLR4 through K48-linked ubiquitination.
ring finger protein 216spermatogoniaubiquitinationcell movementcell proliferation
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