Fig 2: Screening for downstream target genes of BNC1.(A) Heatmap of gene expression changes after overexpression of BNC1. (B) Venn diagram showing the screening of target genes of BNC1. Genes showing decreased expression after overexpression of BNC1 and highly expressed genes in gastric cancer from the TCGA database. (C) Downstream target genes of BNC1 were screened by transcriptome sequencing in AGS stable cell lines overexpressing BNC1 and control cells (log2 fold change [FC] > 1.5 or < −1.5). (D–H) mRNA levels of PTPRC, SERPINB9, RAC2, BAKN1, and CCL20 in MKN-28, AGS and GES-1 cell lines. (I) KEGG analysis of genes showing decreased expression after overexpression of BNC1. (J) mRNA levels of CCL20 in MKN-28 and AGS stable cell lines overexpressing BNC1 and in control cells. (K) ELISA shows the protein levels of CCL20 in MKN-28 and AGS cell lines after overexpression of BNC1. (L) AlphaFold3 prediction of BNC1 binding to the CCL20 promoter region. (M) Dual luciferase results for BNC1 and CCL20.

Fig 3: CCL20 promotes the development of gastric GC cells.(A) CCK-8 assay of cell growth with CCL20 knockdown and control cells. (B) Colony formation assay and (C) quantitative analysis of CCL20 knockdown and control cells. (D) Wound healing assay and (E) quantitative analysis of CCL20 knockdown and control cells. (F) Migration and invasion assays and (G) quantitative analysis of CCL20 knockdown and control cells. (H) Representative images of CCL20 IHC staining in gastric tumor tissues and adjacent normal tissues, along with (I) quantification of CCL20 scores (n = 37; magnification 200× and 400×). (J) Representative images of CCL20 IHC staining in N0 GC tissues and N3 GC tissues, along with (K) quantification of CCL20 scores (n = 37; magnification 200× and 400×).

Fig 4: BNC1 acts on the CCL20 promoter to suppress the expression of CCL20 in GC cells.CCL20 expression mediates the JAK-STAT signaling pathway, leading to the development and progression of GC cells. Created with BioRender.com.