RNA-seq of HGC27 cells carrying shNC or shDLD.

HGC27 cells were transfected with lentivirus carrying shNC or shDLD. Virus transfection was performed by incubating cells with medium containing lentivirus and 8 ug/mL polybrene (OBio) for 24 hours. Cells were then selected with puromycin (Sigma) until no cells in the empty control group were alive. RNA-seq analysisAfter RNA extraction and library construction, sequencing was conducted on the Illumina platform, generating fastq files. Subsequently, quality control was performed using fastQC, followed by alignment to the human genome hg38 to generate a gene expression matrix. Differential gene expression analysis was carried out using DESeq2, with genes selected based on a p-value less than 0.05. Enrichment analysis was then conducted using the enricher function from the ClusterProfiler package, focusing on HALLMARK, GO_BP, REACTOME, and KEGG gene sets, and the results were visualized using bubble plots. GSEA analysis was performed within gene sets of interest after sorting all genes by log2FoldChange.