POFUT2 Mediated Fucosylation of JUP Enhances VEGFA Expression to Promote Angiogenesis in Colorectal Cancer

TL;DR

This study shows that POFUT2, a glycosyltransferase, promotes angiogenesis in colorectal cancer by fucosylating JUP, which increases VEGFA levels.

Contribution

The novel finding is that POFUT2 enhances CRC angiogenesis via fucosylation of JUP, linking glycosylation to VEGFA expression and prognosis.

Findings

POFUT2 is upregulated in CRC tissues and correlates with poor prognosis.

POFUT2 fucosylates JUP, increasing its expression and promoting VEGFA and angiogenesis.

JUP and AGO2 positively regulate angiogenesis, while HSBP1 negatively regulates it.

Abstract

Background: Colorectal cancer (CRC) ranks as the third most common cancer globally and is characterized by a poor prognosis. Abnormal glycosylation, a hallmark of cancer development, influences multiple signaling pathways and contributes to CRC progression. Identifying key glycosyltransferase genes associated with CRC prognosis could provide novel therapeutic targets and improve patient outcomes.

Methods: We utilized datasets from The Cancer Genome Atlas (TCGA) to analyze the expression of glycosyltransferase genes in CRC tissues. Lasso regression and COX regression models were employed to identify key glycosyltransferase genes associated with patient prognosis. Angiogenesis assays were performed utilizing tumor-conditioned medium to assess the influence of GDP-fucose protein O-fucosyltransferase 2 (POFUT2) in cancer cells on human umbilical vein endothelial cells (HUVECs).Flag-Immunoprecipitation combining mass spectrometry detection was employed to identify potential interacting proteins with POFUT2.Western blot, immunoprecipitation, and immunohistochemistry were used to assess the interaction between POFUT2 and Junction Plakoglobin (JUP), as well as the correlation between the expression of Vascular endothelial growth factor A (VEGFA) and Platelet Endothelial Cell Adhesion Molecule-1 (CD31).

Results: Our analysis revealed that POFUT2 is significantly upregulated in CRC tissues and correlates with poor prognosis. Elevated POFUT2 expression in CRC cells enhances proliferation, invasion, and angiogenic capabilities of HUVECs. Among POFUT2 potential interacting proteins, three proteins were found to be involved in angiogenesis: JUP, HSBP1 (Heat Shock Factor Binding Protein 1), and AGO2 (Argonaute RISC Catalytic Component 2). Specifically, HSBP1 negatively regulates angiogenesis, whereas JUP and AGO2 positively regulate angiogenesis. Our results demonstrated that POFUT2 promotes the protein expression of JUP but does not affect the expression of AGO2. Further investigations revealed that POFUT2 interacts with JUP, upregulates its expression through fucosylation, and subsequently regulates VEGFA levels, thereby enhancing angiogenesis. A significant positive correlation was observed between POFUT2 and the expression levels of JUP, VEGFA, and CD31 in CRC tissues.

Conclusions: POFUT2 is identified as a critical glycosyltransferase gene in CRC, closely associated with angiogenic phenotypes and poor prognosis. High POFUT2 expression in CRC regulates JUP fucosylation, increasing JUP and VEGFA levels, which promotes angiogenesis. These findings suggest POFUT2 could serve as a prognostic marker for colorectal cancer, and targeting it may inhibit angiogenesis and aid in treatment.

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