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Promotion of cell migration by neural cell adhesion molecule (NCAM) is enhanced by PSA in a polysialyltransferase-specific manner.

Guan F, Wang X, He F - PLoS ONE (2015)

Bottom Line: We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7.The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied.We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.

ABSTRACT
Neural cell adhesion molecule 140 (NCAM-140) is a glycoprotein and always highly polysialylated in cancer. Functions of polysialic acid (PSA) that binds to N-glycan termini on NCAM remain unclear. ldlD-14 cells, a CHO cell mutant deficient in UDP-Gal 4-epimerase, are useful for structural and functional studies of Gal-containing glycoproteins because their abnormal glycosylation can be converted to normal status by exogenous addition of galactose (Gal). We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7. The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied. We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility. In addition, PSA catalyzed by different polysialyltransferases affected the adhesion of NCAM to different extracellular matrix (ECM) components.

No MeSH data available.


Related in: MedlinePlus

Overexpression of NCAM-140 in ldlD-14 cells.(A) Western blot analysis with β-tubulin used as loading control. (B) Immunofluorescence staining of NCAM and PSA-NCAM. ldlD-14 and ldlD/N140 cells were cultured in serum-free medium supplemented with 5% FBS or ITS or ITS+Gal, and nuclei were visualized by Hoechst staining. Size bars: 20 μm.
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pone.0124237.g001: Overexpression of NCAM-140 in ldlD-14 cells.(A) Western blot analysis with β-tubulin used as loading control. (B) Immunofluorescence staining of NCAM and PSA-NCAM. ldlD-14 and ldlD/N140 cells were cultured in serum-free medium supplemented with 5% FBS or ITS or ITS+Gal, and nuclei were visualized by Hoechst staining. Size bars: 20 μm.

Mentions: To investigate the function of NCAM-140 and PSA-NCAM, the NCAM-140 gene from NMuMG cells was cloned. NCAM levels in parental ldlD-14 cells were undetectable, whereas transfected cells (ldlD/N140) showed a strong NCAM-140 signal in all tested conditions (Fig 1A and 1B).


Promotion of cell migration by neural cell adhesion molecule (NCAM) is enhanced by PSA in a polysialyltransferase-specific manner.

Guan F, Wang X, He F - PLoS ONE (2015)

Overexpression of NCAM-140 in ldlD-14 cells.(A) Western blot analysis with β-tubulin used as loading control. (B) Immunofluorescence staining of NCAM and PSA-NCAM. ldlD-14 and ldlD/N140 cells were cultured in serum-free medium supplemented with 5% FBS or ITS or ITS+Gal, and nuclei were visualized by Hoechst staining. Size bars: 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401701&req=5

pone.0124237.g001: Overexpression of NCAM-140 in ldlD-14 cells.(A) Western blot analysis with β-tubulin used as loading control. (B) Immunofluorescence staining of NCAM and PSA-NCAM. ldlD-14 and ldlD/N140 cells were cultured in serum-free medium supplemented with 5% FBS or ITS or ITS+Gal, and nuclei were visualized by Hoechst staining. Size bars: 20 μm.
Mentions: To investigate the function of NCAM-140 and PSA-NCAM, the NCAM-140 gene from NMuMG cells was cloned. NCAM levels in parental ldlD-14 cells were undetectable, whereas transfected cells (ldlD/N140) showed a strong NCAM-140 signal in all tested conditions (Fig 1A and 1B).

Bottom Line: We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7.The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied.We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.

ABSTRACT
Neural cell adhesion molecule 140 (NCAM-140) is a glycoprotein and always highly polysialylated in cancer. Functions of polysialic acid (PSA) that binds to N-glycan termini on NCAM remain unclear. ldlD-14 cells, a CHO cell mutant deficient in UDP-Gal 4-epimerase, are useful for structural and functional studies of Gal-containing glycoproteins because their abnormal glycosylation can be converted to normal status by exogenous addition of galactose (Gal). We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7. The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied. We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility. In addition, PSA catalyzed by different polysialyltransferases affected the adhesion of NCAM to different extracellular matrix (ECM) components.

No MeSH data available.


Related in: MedlinePlus