Human colon cancer cell lines permanently expressing alpha 2,6-sialylated sugar chains by transfection with rat beta-galactoside alpha 2,6 sialyltransferase cDNA.
A role of alpha 2,6-linked sialic acid in the development of an invasive phenotype in colon cancer has been suggested by several observations but never conclusively demonstrated. An experimental model to clarify this tissue was established by the creation and characterization of a bank of cell lines that differ mainly, if not exclusively, in the degree of alpha 2,6-sialylation. Human colon cancer cell lines SW48 and SW948, normally unable to elaborate the alpha 2,6-sialyl linkage, were transfected with the beta-galactoside alpha 2,6-sialyltransferase (alpha 2,6ST) cDNA driven by the cytomegaloviral promoter and screened for cell surface alpha 2,6-sialylated sugar chains using fluorescein isothiocyanate-conjugated Sambucus nigra agglutinin (SNA-FITC). A panel of SNA-FITC-positive clones was established that expresses alpha 2,6ST activity of varying degrees. Only the SNA-FITC-positive recombinants express the 1.2 Kb mRNA predicted to be generated from the transfected sequence. No 4.3-4.7 Kb transcripts that are indicative of transcription from the native alpha 2,6ST gene were detected.
Dall'Olio
F
F
Dipartimento di Pathologia Sperimentale, Università degli Studi di Bologna, Italy.
Chiricolo
M
M
Lollini
P
P
Lau
J T
JT
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003110:Colonic Neoplasms; D003587:Cytomegalovirus; D017931:DNA Primers; D018076:DNA, Complementary; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006001:Glycoconjugates / Q000032:analysis / Q000096:biosynthesis; D006801:Humans; D037102:Lectins; D008969:Molecular Sequence Data; D037121:Plant Lectins; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D054788:Ribosome Inactivating Proteins; D012794:Sialic Acids / Q000032:analysis*; D012799:Sialyltransferases / Q000096:biosynthesis / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D017931:DNA Primers; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D006001:Glycoconjugates; D037102:Lectins; D037121:Plant Lectins; D012333:RNA, Messenger; D011994:Recombinant Proteins; C050600:Sambucus nigra lectins; D012794:Sialic Acids; D012799:Sialyltransferases; D054788:Ribosome Inactivating Proteins; D016650:Fluorescein-5-isothiocyanate; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase