A mucus-secreting human colonic cancer cell line. Purification and partial characterization of the secreted mucins.
A stably differentiated clonal derivative (Cl.16E) of the human colonic adenocarcinoma cell line HT29 secretes in culture high-Mr glycoproteins that were purified from the serum-free conditioned medium by preparative SDS/polyacrylamide-gel electrophoresis. Analysis of the oligosaccharides released from the [3H]glucosamine-labelled high-Mr glycoproteins by alkaline-borohydride treatment showed that this material consisted of O-linked oligosaccharides (without any detectable N-linked oligosaccharides) that were eluted as three fractions from Bio-Gel P-6 columns. The main oligosaccharide fraction obtained after such treatment and desialylation was eluted together with a six-unit glucose polymer from a Bio-Gel P-4 column. Polyclonal antibodies were raised against the high-Mr glycoproteins, and in immunoblot analysis they reacted specifically with the high-Mr glycoproteins present in the conditioned medium. Furthermore, immunohistochemical staining of sections in paraffin wax revealed that these antibodies labelled normal human gastrointestinal mucins. We conclude that (1) the high-Mr glycoproteins prepared by SDS/polyacrylamide-gel electrophoresis are pure mucus glycoproteins on the basis of sensitivity to alkaline-borohydride treatment, monosaccharide composition and immunochemical and immunohistological findings, and (2) these mucins have antigenic determinants in common with the normal human gastrointestinal mucins.
Maoret
J J
JJ
Laboratoire de Biologie et Physiologie des Cellules Digestives, Unité 239 I.N.S.E.R.M., Faculté de Médecine Bichat, Paris, France.
Font
J
J
Augeron
C
C
Codogno
P
P
Bauvy
C
C
Aubery
M
M
Laboisse
C L
CL
D000230:Adenocarcinoma / Q000378:metabolism*; D002460:Cell Line; D002850:Chromatography, Gel; D003110:Colonic Neoplasms / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D009077:Mucins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism
D009077:Mucins