VGLL4 promotes hepatocellular carcinoma progression via the Wnt/β-catenin pathway

Hepatocellular carcinoma (HCC) has a poor prognosis owing to its late diagnosis and limited therapies. This study aimed to clarify the role and mechanism of action of VGLL4 in the progression of HCC. We analyzed RNA-seq data and clinical information from 355 patients with HCC and 43 non-tumor controls in The Cancer Genome Atlas (TCGA) cohort and performed in vitro and in vivo experiments. TCGA mining revealed that VGLL4 was significantly upregulated in HCC tumors compared to normal tissues, with its expression escalating with TNM stage and independent of sex and age. High VGLL4 expression independently predicted shorter overall survival and progression-free survival (hazard ratio associated with poor prognosis) and exhibited diagnostic value. In vitro, VGLL4 overexpression in HuH-7 HCC cells increased proliferation (measured by the CCK-8 assay), migration (wound healing assay), invasion (Transwell assay), and stemness (tumor sphere formation assay). In vivo, subcutaneous xenografts of VGLL4-overexpressing HuH-7 cells (4 ×10⁶ cells inoculated into 4-week-old male BALB/c nude mice and observed for 8 weeks) showed larger tumor volumes and heavier weights than controls. Mechanistically, VGLL4 activated the Wnt/β-catenin pathway (upregulated nuclear β-catenin, TCF4, and c-Myc), induced epithelial-mesenchymal transition (EMT, upregulating vimentin and N-cadherin), and was positively correlated with stromal infiltration by cancer-associated fibroblasts (CAFs), myeloid-derived suppressor cells (MDSCs), macrophages, and endothelial cells in the TCGA cohort. Thus, VGLL4 drives HCC progression by linking tumor-intrinsic EMT/stemness programs and stromal crosstalk via the Wnt/β-catenin pathway, serving as a prognostic biomarker and a potential therapeutic target.