Abstract
Liver cancer ranks sixth in incidence and third in mortality worldwide, with hepatocellular carcinoma (HCC) accounting for 90% of cases. Kinesin family genes (KIFs) have been implicated in HCC progression, but their specific roles remain unclear. Here, using multiple transcriptomic datasets and machine learning, we systematically screened KIF genes in HCC and identified nine key differentially expressed KIFs (KIF3A, KIF3B, KIF5B, KIF11, KIF13A, KIF13B, KIF20A, KIFC3, KLC2) as diagnostic biomarkers. A logistic regression model showed excellent diagnostic accuracy (AUC = 0.992) in the GSE121248 dataset. Six genes (KIF3A, KIF5B, KIF11, KIF13B, KIF20A, KLC2) were validated in TCGA, several correlating with prognosis. Notably, KIF20A was an independent prognostic factor, validated across cohorts. Single-cell and experimental data revealed high KIF20A expression in HCC cells, promoting proliferation and migration. Mechanistically, KIF20A drives tumor growth and metastasis via Wnt/β-catenin signaling and epithelial-mesenchymal transition (EMT) proteins such as N-cadherin, Slug, Snail, and Twist1/2. Wnt pathway activator SKL2001 and inhibitor LGK974 confirmed KIF20A's role in tumor progression. Upstream, the transcription factor FOXK1 was identified as a key positive regulator of KIF20A. FOXK1 is overexpressed in HCC, strongly correlates with KIF20A, and predicts poor prognosis. ChIP-seq and promoter assays verified FOXK1's direct binding to the KIF20A promoter, activating its transcription. In conclusion, KIF20A serves as a diagnostic and prognostic biomarker promoting HCC progression via Wnt/β-catenin signaling, regulated by FOXK1, offering new therapeutic targets.