Transgene expression of alpha(1,2)-fucosyltransferase-I (FUT1) in tumor cells selectively inhibits sialyl-Lewis x expression and binding to E-selectin without affecting synthesis of sialyl-Lewis a or binding to P-selectin.
During inflammation, E- and P-selectins appear on activated endothelial cells to interact with leukocytes through sialyl-Lewis x and sialyl-Lewis a antigens (sLe(x/a)). These selectins can also interact with tumor cells in a sialyl-Lewis-dependent manner and for this reason, they are thought to play a key role in metastasis. Diverting the biosynthesis of sialyl-Lewis antigens toward nonadhesive structures is an attractive gene therapy for preventing the hematogenous metastatic spread of cancers. We have previously shown that transfection of alpha(1,2)-fucosyltransferase-I (FUT1) in Chinese hamster ovary (CHO) cells had a slight effect on the overall sialylation while the synthesis of sLE(x) was dramatically prevented. We herein delivered the gene of FUT1 by a human immunodeficiency virus-derived lentiviral vector to three human cancer cell lines including pancreatic (BxPC3), hepatic (HepG2), and colonic (HT-29) cancer cells. We found that on FUT1 transduction, all cells exhibited a dramatic decrease in sLe(x) synthesis with a concomitant increase in Le(y) and Le(b) expression, without any detectable effect on the level of cell surface sLe(a) antigens. In parallel, FUT1-transduced HT-29 and HepG2 cells, but not BxPC3 cells, failed to interact with E-selectin as assessed by E-selectin-binding assay or dynamic adhesion to activated endothelial cells. We show also that transduced FUT1 efficiently fucosylates the P-selectin ligand PSGL-1 without altering P-selectin binding. These results have important implications for understanding cell-specific reactions underlying the synthesis of selectin ligands in cancer cells and may provide a basis for the development of anti-metastatic gene therapy.
Mathieu
Sylvie
S
Faculté de Médecine, INSERM U-559/UEA-3289, Marseille, France.
Prorok
Maëlle
M
Benoliel
Anne-Marie
AM
Uch
Rathviro
R
Langlet
Claire
C
Bongrand
Pierre
P
Gerolami
René
R
El-Battari
Assou
A
D000818:Animals; D018395:CA-19-9 Antigen; D002448:Cell Adhesion / Q000502:physiology; D019040:E-Selectin / Q000378:metabolism*; D042783:Endothelial Cells / Q000502:physiology; D005434:Flow Cytometry; D005453:Fluorescence; D005647:Fucosyltransferases / Q000096:biosynthesis* / Q000235:genetics; D005732:Gangliosides / Q000378:metabolism; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics; D019073:HT29 Cells; D006801:Humans; D015151:Immunoblotting; D008562:Membrane Glycoproteins / Q000378:metabolism; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen; D014161:Transduction, Genetic; D014162:Transfection; D019076:Transgenes
D018395:CA-19-9 Antigen; D019040:E-Selectin; D005732:Gangliosides; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D019007:P-Selectin; C084882:P-selectin ligand protein; D000080506:Sialyl Lewis X Antigen; C064564:sialyl Le(a) ganglioside; D005647:Fucosyltransferases