Exploring the combined roles of GALNT1 and GALNT2 in hepatocellular carcinoma malignancy and EGFR modulation.

Abstract

Background: Hepatocellular carcinoma (HCC), the most formidable subtype of primary liver cancers, is becoming increasingly concerning due to its rising incidence worldwide. HCC ranks as the sixth most diagnosed cancer globally and is the third leading cause of cancer-related deaths. Glycosylation, a common post-translational modification of proteins, is frequently altered in tumors and is associated with the progression of malignancies. GALNT1 and GALNT2 are GalNAc-transferases that initiate protein O-glycosylation and are closely linked to cancer development. Investigating the relationship between GALNT1 and GALNT2 in HCC could provide new insights into the disease's pathogenesis. Thus, this study aimed to explore the combined effects of GALNT1 and GALNT2 transfection on HCC, compared to the effects of modifying each gene individually.

Materials and methods: GALNT1 and GALNT2 were assessed by bioinformatics, qPCR, and Western blot analyses to detect their expression in HCC tissues and cell lines. The effects of GALNT1/GALNT2 overexpression and knockdown on cell viability, proliferation, migration, invasion, and apoptosis were evaluated in HCC cells using CCK8, colony formation, transwell migration and invasion, wound healing, TUNEL, and flow cytometry assays. EGFR protein levels were also analyzed by Western blotting.

Results: Co-transfection of GALNT1 knockdown with GALNT2 overexpression significantly suppressed proliferation, migration, and invasion, while promoting apoptosis in HCC cells. Conversely, co-transfection of GALNT1 overexpression with GALNT2 knockdown enhanced these malignant characteristics compared to the modified single gene. Notably, we observed that GALNT1 and GALNT2 modulated EGFR protein expression. Overall, our findings suggest that the combined activity of GALNT1 and GALNT2 is critical in regulating HCC malignant behaviors.