Exploration On The Molecular Characteristics And Functionalities Of Key Genes ATP6V1B1 And TGFBR2 Driving Triple Negative Breast Cancer Stemness

Breast Cancer is a highly heterogeneous disease with significantly different clinical outcomes and Cancer stem cell(CSC)proportions.Compared with other subtypes,Triple negative breast cancer(TNBC)has the highest degree of malignancy and a higher proportion of CSC,so there is no effective treatment.The latest research indicates that CSC targeted therapy is expected to be a breakthrough in the treatment of TNBC.Therefore,in-depth study of the CSC molecular characteristics of TNBC is helpful to resolve the heterogeneity of breast cancer and provide a new idea for the construction of TNBC treatment targeting CSC.Objective: To study the molecular characteristics of CSC in TNBC,search for potential biomarkers and therapeutic targets,explore its mechanism,and provide ideas and theoretical basis for CSC targeted therapy in TNBC.Methods: The CSC transcriptome data of TNBC were analyzed for gene differential expression.Genomic Ontology(GO)and the Kyoto Encyclopedia of Genes and Genomes Genomes(KEGG)enrichment analysis was used to find the biological functions and signaling pathways involved."Extracellular matrix tissue" and "epidermal development" ranked first in the up-regulation and down-regulation enrichment results of GO,and "PI3K/AKT signaling pathway" and "cell tight connection" ranked first in the up-regulation and down-regulation enrichment results of KEGG.Key genes were searched by protein regulatory network construction method.Small interfering RNA was used to knock down the expression of ATP6V1B1 in breast cancer cell line SKBR3.The effect of low expression of ATP6V1B1 on CSC ratio of SKBR3 was detected by cell acetaldehyde dehydrogenase,and the effect of low expression of ATP6V1B1 on cell invasion ability was detected by transwell assay.The expression of TGFBR2 in TNBC cell line SUM159 PT was knocked down by small interfering RNA,and the effect of TGFBR2 on cell metastasis was detected by scratch and transwell assay.Cold atmospheric plasma(CAP)was used to treat TNBC cell line SUM159 PT,and Western-blot was used to detect its effect on TGFBR2 expression.Proteomics,mass spectrometry and immunoprecipitation methods were used to investigate the mechanism of CAP targeting TGFBR2 to inhibit cell metastasis.Results: A CSC gene combination of TNBC was constructed,which consisted of 122 high expression and 381 low expression genes.Key genes ATP6V1B1 and TGFBR2 were selected through the protein regulatory network.After knocking down ATP6V1B1 in breast cancer cell line SKBR3,the proportion of CSC was significantly reduced,and the ability of invasion was significantly weakened.In TNBC cell line SUM159 PT,knockdown gene TGFBR2 could effectively inhibit cell metastasis.In TNBC cell line SUM159 PT,CAP treatment decreased the expression of TGFBR2 and inhibited cell metastasis by SASH1-HSP90 axis,which significantly increased E-CAD and decreased VIM.Conclusion:(1)CSC gene combinations can represent most of the CSC molecular characteristics of TNBC,and are closely related to cancer metastasis.(2)Knocking down ATP6V1B1 can effectively reduce the proportion of CSC in breast cancer cell line SKBR3 and inhibit cell invasion.(3)Knockdown of TGFBR2 can effectively reduce the cell metastasis ability of TNBC cell line SUM159 PT.(4)CAP treatment reduced the expression of TGFBR2 through the SASH1-HSP90 axis and inhibited the transfer ability of TNBC cell line SUM159 PT and epithelial mesenchymal transformation process.