S100A10在三阴性乳腺癌中的促癌机制研究OA

AIM:To investigate the effects of S100 calcium-binding protein A10(S100A10)on cell viability,migration,colony-forming capacity,apoptosis,cell cycle progression,and tumorigenicity of triple-negative breast cancer(TNBC)cells in vivo,as well as to explore the underlying molecular mechanisms.METHODS:The human TNBC cell lines SUM159PT and MDA-MB-231 were employed for functional characterization in vitro.The stable cell models of S100A10 knockdown and overexpression were established via lentiviral transduction and subsequent antibiotic selection in these cells.Cell viability was quantified by the CCK-8 assay,clonogenic potential was evaluated by colony formation as-say,migratory capacity was assessed via wound-healing and Transwell migration assays,cell cycle distribution and apopto-sis rates were determined by flow cytometry.The expression levels of apoptosis-related proteins,including pro-apoptotic Bax and anti-apoptotic Bcl-2,were examined by Western blot.To evaluate the impact of S100A10 on TNBC tumorigenici-ty in vivo,MDA-MB-231 cells with stable S100A10 knockdown were subcutaneously injected into immunodeficient NCG mice.RNA sequencing was performed on S100A10-knockdown versus control MDA-MB-231 cells to identify differentially expressed genes and conduct pathway enrichment analysis.Western blot was conducted to verify the expression of NF-κB signaling pathway-related proteins.Finally,a dual-luciferase reporter assay was conducted to determine whether the key NF-κB transcription factor RelA directly binds to and transactivates the S100A10 promoter.RESULTS:Functional as-says in vitro revealed that S100A10 knockdown significantly suppressed viability,migration and clonogenic capacity in TN-BC cells,induced G0/G1-phase cell cycle arrest,upregulated the pro-apoptotic protein Bax,downregulated the anti-apop-totic protein Bcl-2 and increased apoptotic cell death(all P<0.05).In contrast,S100A10 overexpression consistently re-versed these phenotypes(all P<0.05).In vivo,stable S100A10 knockdown markedly suppressed the growth of subcutane-ous xenograft tumors in immunodeficient NCG mice(P<0.05).The results of RNA sequencing analysis revealed that dif-ferentially expressed genes following S100A10 knockdown were significantly enriched in the NF-κB signaling pathway.Consistent with this,Western blot analysis further demonstrated that S100A10 knockdown significantly reduced phosphory-lation of the NF-κB subunit P65,whereas S100A10 overexpression enhanced P65 phosphorylation(both P<0.05).Fur-thermore,dual-luciferase reporter assays confirmed that the NF-κB transcription factor RelA directly transactivated the S100A10 promoter,suggesting that the NF-κB signaling pathway might regulate S100A10 expression at the transcriptional level.CONCLUSION:In TNBC,S100A10 can promote cell viability,migration,inhibit cell apoptosis,and accelerate tumor growth,which may be related to the S100A10/NF-κB loop.