Expression of mucin-associated tumor antigens is altered by cell density.
Mucin-associated sialylated Lewis antigens are implicated in tumor cell metastasis and are used in several tests for pancreatic cancer. Despite their clinical importance, little is known about the structures of the oligosaccharides of pancreatic cancer mucins or about the regulation of their synthesis or of the synthesis of their protein cores. In this study, we examined the effects of culture at high cell density on the expression of these antigens in the SW1990 human pancreatic cancer cell line. Mucins from cells that were 2.5 weeks post-confluent had increased expression of sialyl-Lewis(a) and Lewis(x) antigens but reduced expression of the DU-PAN-2 antigen (NeuAc alpha2,3Galbeta1,3GlcNAc-Gal-R) when compared to mucins from 1 day post-confluent cells. Sialyl-Lewis antigens differ from the DU-PAN-2 antigen by the presence of an additional fucose. Mucins from 2.5-week cells also had increased binding to lectins specific for fucose, such as AAL and UEAI, with no apparent change in the binding of lectins specific for sialic acids. Metabolically radiolabeled O-linked oligosaccharides with sialyl-Lewis(a) antigenic reactivity eluted from Bio-Gel P-10 in the region of sialylated and sulfated oligosaccharides. Oligosaccharides eluted from QAE-Sephadex (2 mM Tris base) in a pattern suggesting the presence of 1, 2 and 3 or more negative charges per oligosaccharide. Even after desialylation and desulfation, oligosaccharides eluted from Bio-Gel P-10 with apparent molecular sizes greater than glucose oligomers of 12 units. Culture of SW 1990 cells at high density also increased the steady-state levels of mRNA for mucins MUC1, 2, 4, 5 and 6. In summary, after prolonged culture at high cell density, SW1990 cells have qualitative changes in their oligosaccharides that may be due to up-regulation of fucosyltransferases.
Yamashita
Y
Y
First Department of Surgery, Osaka City University Medical School, Osaka, Japan.
Ho
J J
JJ
Cheng
S
S
Siddiki
B
B
Chung
Y S
YS
Sowa
M
M
Kim
Y S
YS
D015295:Antigens, Tumor-Associated, Carbohydrate / Q000032:analysis* / Q000302:isolation & purification; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002452:Cell Count; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D004563:Electrochemistry; D006801:Humans; D008969:Molecular Sequence Data; D009077:Mucins / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D010190:Pancreatic Neoplasms / Q000276:immunology* / Q000473:pathology*; D012333:RNA, Messenger / Q000032:analysis; D013997:Time Factors; D014407:Tumor Cells, Cultured
D015295:Antigens, Tumor-Associated, Carbohydrate; D009077:Mucins; D009844:Oligosaccharides; D012333:RNA, Messenger; C058981:sialyl-Le(a) oligosaccharide