Polysialic acid (
polySia), an α-2,8-glycosidically linked
polymer of
sialic acid, is a developmentally regulated post-translational modification predominantly found on
NCAM (neuronal
cell adhesion molecule). Whilst high levels are expressed during development, peripheral adult organs do not express
polySia-
NCAM. However, tumours of neural crest-origin re-express
polySia-
NCAM: its occurrence correlates with aggressive and invasive disease and poor clinical prognosis in different
cancer types, notably including
small cell lung cancer (SCLC),
pancreatic cancer and
neuroblastoma. In neuronal development,
polySia-
NCAM biosynthesis is catalysed by two polysialyltransferases, ST8SiaII and ST8SiaIV, but it is ST8SiaII that is the prominent
enzyme in tumours. The aim of this study was to determine the effect of ST8SiaII inhibition by a small molecule on tumour cell migration, utilising
cytidine monophosphate (
CMP) as a tool compound. Using immunoblotting we showed that
CMP reduced ST8iaII-mediated polysialylation of
NCAM. Utilizing a novel HPLC-based assay to quantify polysialylation of a fluorescent acceptor (DMB-DP3), we demonstrated that
CMP is a competitive inhibitor of ST8SiaII (K i = 10 µM). Importantly, we have shown that
CMP causes a concentration-dependent reduction in tumour cell-surface
polySia expression, with an absence of toxicity. When ST8SiaII-expressing tumour cells (SH-SY5Y and C6-STX) were evaluated in 2D cell migration assays, ST8SiaII inhibition led to significant reductions in migration, while
CMP had no effect on cells not expressing ST8SiaII (DLD-1 and C6-WT). The study demonstrates for the first time that a polysialyltransferase inhibitor can modulate migration in ST8SiaII-expressing tumour cells. We conclude that ST8SiaII can be considered a druggable target with the potential for interfering with a critical mechanism in tumour cell dissemination in metastatic
cancers.