Effect of TRIM21 on influencing cellular behaviors of the breast cancer cell line MCF-7 via the glutamine metabolism

HU Yanglin; HE Xiongbin; HE Jianhuai

doi:10.3969/j.issn.1002-7386.2025.10.001

Abstract：Objective To investigate the effect of tripartite motif protein 21(TRIM21)on the biological behavior of human breast cancer MCF-7 cells by interfering with glutamine metabolism.Methods The human breast cancer cell line MCF-7 and human normal breast epithelial cell line MCF-10A were purchased.Protein expression of TRIM21 in MCF-7 and MCF-10A cells was detected by Western blot.MCF-7 cells were induced with blank control,or transfected with pcDNA3.1-NC or pcDNA3.1-TRIM21.The proliferation,invasion,and migration abilities of MCF-7 cells were detected by MTT assay,Transwell assay,and wound healing assay,respectively.Flow cytometry was used to detect the cell cycle and apoptosis rate of MCF-7 cells.Glutamine(Gln),glutathione(GSH)and glutamate(Glu)levels in MCF-7 cells were measured using enzyme-linked immunosorbent assay(ELISA).Protein levels of TRIM21,glutamate dehydrogenase 1(GLUD1),and glutaminase 1(GLS1)in MCF-7 cells were detected by Western blot.Results Compared with that in MCF-10A cells,TRIM21 was significantly downregulated in MCF-7 cells(P＜0.05).Transfection of pcDNA3.1-TRIM21 significantly increased the apoptotic rate and protein level of TRIM21 in MCF-7 cells,but decreased invasion rate,migratory rate,contents of Gln,Glu and GSH,and protein levels of GLS1 and GLUD1(P＜0.05).Transfection of pcDNA3.1-TRIM21 significantly increased cell number in S phase,and decreased that in G0/G1 and G2/M phases(P＜0.05).Cell proliferative rate at 24 h,48 h and 72 h significantly decreased in MCF-7 cells transfected with pcDNA3.1-TRIM21(P＜0.05).Conclusion TRIM21 is lowly expressed in breast cancer,and overexpression of TRIM21 can inhibit the proliferation,invasion and migration of breast cancer cells,arrest cell cycle in S phase,and induce apoptosis by inhibiting glutamine metabolism.