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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

Cell adhesion assay.ldlD-14 (mock), ldlD/plasmid (control), ldlD/N140, ldlD/N/S, ldlD/N/P cells were cultured in 5% FBS medium for 48 hr. Medium was replaced with ITS or ITS+Gal medium and cultured for 48 hr. Cell adhesion to FN, LN, collagen IV, matrigel or BSA solution was determined as described in Materials and Methods. The absorbance of fixed and crystal-violet stained cells was recorded at 595 nm. Four independent experiments gave similar results. *, p<0.05; **, p = 0.01–0.05; ***, p<0.01 vs. control.
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pone.0124237.g006: Cell adhesion assay.ldlD-14 (mock), ldlD/plasmid (control), ldlD/N140, ldlD/N/S, ldlD/N/P cells were cultured in 5% FBS medium for 48 hr. Medium was replaced with ITS or ITS+Gal medium and cultured for 48 hr. Cell adhesion to FN, LN, collagen IV, matrigel or BSA solution was determined as described in Materials and Methods. The absorbance of fixed and crystal-violet stained cells was recorded at 595 nm. Four independent experiments gave similar results. *, p<0.05; **, p = 0.01–0.05; ***, p<0.01 vs. control.

Mentions: Literature shows that altered cell adhesion is involved in tumor metastasis and progression, and that the extracellular matrix (ECM) plays an important role in the regulation of cell adhesion[38]. We further examined the effect of NCAM and PSA on the attachment of ldlD-14 cells to the following ECM components: fibronectin (FN), laminin (LN), collagen IV and matrigel (Fig 6). ldlD-14 cells were more adherent to LN and collagen IV than FN and matrigel. In ITS condition, NCAM-140 overexpressing cells (ldlD/N140, ldlD/N/S and ldlD/N/P) were highly adherent to FN and poorly adherent to LN, collagen IV and matrigel, compared with ldlD-14 cells. These different adhesive properties became more pronounced in ITS+Gal condition. ldlD/N/S and ldlD/N/P cells presented decreased adhesion to FN and collagen IV, and increased adhesion to matrigel, compared to ldlD/N140 cells. Interestingly, adhesion to LN was not changed in ldlD/N/S cells, but greatly increased in ldlD/N/P cells. These results suggested that PSA catalyzed by different polysialyltransferases affected the adhesion of NCAM to ECM components.


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)

Cell adhesion assay.ldlD-14 (mock), ldlD/plasmid (control), ldlD/N140, ldlD/N/S, ldlD/N/P cells were cultured in 5% FBS medium for 48 hr. Medium was replaced with ITS or ITS+Gal medium and cultured for 48 hr. Cell adhesion to FN, LN, collagen IV, matrigel or BSA solution was determined as described in Materials and Methods. The absorbance of fixed and crystal-violet stained cells was recorded at 595 nm. Four independent experiments gave similar results. *, p<0.05; **, p = 0.01–0.05; ***, p<0.01 vs. control.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0124237.g006: Cell adhesion assay.ldlD-14 (mock), ldlD/plasmid (control), ldlD/N140, ldlD/N/S, ldlD/N/P cells were cultured in 5% FBS medium for 48 hr. Medium was replaced with ITS or ITS+Gal medium and cultured for 48 hr. Cell adhesion to FN, LN, collagen IV, matrigel or BSA solution was determined as described in Materials and Methods. The absorbance of fixed and crystal-violet stained cells was recorded at 595 nm. Four independent experiments gave similar results. *, p<0.05; **, p = 0.01–0.05; ***, p<0.01 vs. control.
Mentions: Literature shows that altered cell adhesion is involved in tumor metastasis and progression, and that the extracellular matrix (ECM) plays an important role in the regulation of cell adhesion[38]. We further examined the effect of NCAM and PSA on the attachment of ldlD-14 cells to the following ECM components: fibronectin (FN), laminin (LN), collagen IV and matrigel (Fig 6). ldlD-14 cells were more adherent to LN and collagen IV than FN and matrigel. In ITS condition, NCAM-140 overexpressing cells (ldlD/N140, ldlD/N/S and ldlD/N/P) were highly adherent to FN and poorly adherent to LN, collagen IV and matrigel, compared with ldlD-14 cells. These different adhesive properties became more pronounced in ITS+Gal condition. ldlD/N/S and ldlD/N/P cells presented decreased adhesion to FN and collagen IV, and increased adhesion to matrigel, compared to ldlD/N140 cells. Interestingly, adhesion to LN was not changed in ldlD/N/S cells, but greatly increased in ldlD/N/P cells. These results suggested that PSA catalyzed by different polysialyltransferases affected the adhesion of NCAM to ECM components.

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