Structure of O-glycan Core 2

The formation of the core 2 O-glycan structure relies on three specific glycosyltransferases: C2GnT-1 to 3. These enzymes are responsible for introducing N-acetylglucosamine β 1,6 branching. The multifaceted functions of these glycosyltransferases are associated with various cellular processes. C2GnT-1 stands as a crucial orchestrator in the regulation of lymphocyte rolling, a vitally important phenomenon in the intricate immune response. Moreover, the activation of T-cells exerts a profound impact on C2GnT activity, with its levels being significantly upregulated in response to T-cell activation. Furthermore, the enigmatic C2GnT-3, selectively expressed in activated T-cells, has been found to exert a significant influence on the circulation of thyroxine. The core 2 O-glycans and their glycosyltransferases play a key role in unraveling the intricate mechanisms governing cellular behavior.

Fig.1 The structure of O-glycan core 2. (CD BioGlyco)

O-glycan Core 2 Glycoengineering Service at CD BioGlyco

• CD BioGlyco generates prostate cancer cells that express O-glycan core 2 to determine its role in prostate cancer progression. We stably transfect the LNCaP prostate cancer cell line with cDNA encoding Core2GnT. The cells are being transfected with pcDNA3-Core2GnT using LipofectAMINE. The parent LNCaP cells do not express Core2GnT.
• After the transfection of Core2GnT cDNA, we identify five clones of transfected cells expressing Core2GnT (LNCaP-Core2GnT). In order to establish a control group, we currently perform transfection of LNCaP cells with the pcDNA3 empty vector, resulting in the designation of the transfected cells as LNCaP mock cells.
• Because Immunohistochemical detection of Core2GnT is more efficient compared to the detection of Core2GnT transcripts using in situ hybridization or core 2-branched O-glycans, we assess by staining with anti-Core2GnT antibodies.
• We prepare total RNA using the TRIzol reagent (Invitrogen) and perform reverse transcriptase-polymerase chain reaction (RT-PCR) analysis on glyceraldehyde-3-phosphate dehydrogenase and Core2GnT. The enzymatic activity of Core2GnT is measured by using Gal β1→3GalNAc ɑ1→p-nitrophenol as a substrate.

These findings demonstrate the successful introduction and expression of O-glycan core 2 in LNCaP cells, providing a foundation for further investigations into the functional role of Core2GnT and O-glycan core 2 in prostate cancer progression.

Applications

Advantages

• We use advanced methodologies to investigate the impact of Core2GnT expression and core 2 O-glycans on tumor formation and cell adhesion.
• Our in vivo approach using mouse prostate inoculation allows us to closely mimic the tumor microenvironment found in humans. This enhances the translational value of our research, ultimately contributing to the development of more effective cancer treatments.
• Our team can design robust experimental models and conduct rigorous data analysis.

CD BioGlyco has established itself as a pioneer in the field of glycoengineering, garnering widespread recognition for its innovative methodologies and expertise. Having an exceptional team of distinguished scientists and researchers, we are committed to delivering an extensive suite of Cell Surface Glycoengineering Services that cater to diverse research needs. For any inquiries or to delve further into our services, we warmly welcome you to contact us.