Loss of LLGL1 Elevates EGFR/RAS/MAPK Signaling and Remodels EMT Markers in Huh-7 Hepatocellular Carcinoma Cells

Loss of epithelial polarity is a critical driver of tumor progression; however, how core polarity regulators interface with oncogenic signaling pathways in hepatocellular carcinoma (HCC) remains incompletely defined. LLGL scribble cell polarity complex component 1 (LLGL1) is an evolutionarily conserved polarity protein with well-established tumor-suppressive roles in multiple epithelial malignancies. Nevertheless, how LLGL1 loss shapes oncogenic signaling outputs and cellular phenotypes in HCC remains unclear. In this study, we investigated the consequences of LLGL1 knockout (KO) in epithelial-like Huh-7 HCC cells. LLGL1 loss resulted in enhanced proliferative capacity and increased clonogenic potential, accompanied by altered cell-cycle distribution characterized by reduced G1-phase and increased S-phase fractions (p < 0.001). At the signaling level, LLGL1 KO cells displayed potentiated EGFR-driven RAS/MAPK pathway activation, with increased EGFR phosphorylation, enhanced downstream RAF1-MEK-ERK-RSK signaling, elevated EGFR abundance, and selective modulation of RAF1 protein levels. Functionally, LLGL1 loss markedly enhanced migratory and invasive behavior (p < 0.0001). Despite increased motility, LLGL1 KO cells exhibited remodeling of epithelial-mesenchymal transition (EMT)-associated markers without evidence of a classical EMT program. Collectively, these findings position LLGL1 loss as a central factor associated with altered MAPK signaling, EMT marker remodeling, and tumor-promoting cellular phenotypes in HCC.