ISSN:
1573-4986
Keywords:
sialyltransferases
;
breast cancer cells
;
multiplex RT-PCR
;
glycosyltransferases
;
bp, base pair
;
GAPDH, glyceraldehyde-3-phosphate dehydrogenase
;
kb, kilobase
;
PNA, Peanut (Arachis hypogaea) agglutinin
;
RT-PCR, reverse transcription-polymerase chain reaction
;
TBE, Tris base 0.13 M, boric acid 45 mM
;
Na2EDTA 2.55 mM, pH 8.8 buffer
;
Tm, melting temperature
;
the nomenclature of sialyltransferases is based on that of Tsuji et al. [36]: ST3Gal I: CMP-NeuAc, Gal beta 1-3GalNAc alpha 2,3-sialyltransferase, EC 2.4.99.4
;
ST3Gal III: CMP-NeuAc, Gal beta 1-3/4GlcNAc alpha 2,3-sialyltransferase, EC 2.4.99.6
;
ST3Gal IV: CMP-NeuAc, Gal beta 1-4GlcNAc or Gal beta 1-3GalNAc alpha 2,3-sialyltransferase, EC 2.4.99.
;
ST6Gal I: CMP-NeuAc, Gal beta 1-4GlcNAc alpha 2,6-sialyltransferase, EC 2.4.99.1.
Source:
Springer Online Journal Archives 1860-2000
Topics:
Chemistry and Pharmacology
Notes:
Abstract In many cases of human cancer, the appearance of hypersialylated glycan structures is related to a precise stage of the disease; this may depend on altered regulation of one or more sialyltransferases genes. Since several distinct sialyltransferase enzymes arising from different unique genes transfer sialic acid residues in the same linkage onto the same acceptor, it is impossible to precisely determine which enzyme is involved in the observed phenotype based on enzymatic assays. We have developed a very sensitive and highly reproducible multiplex reverse transcriptase-polymerase chain reaction technique in order to monitor the expression of four human sialyltransferases genes ST6Gal I, ST3Gal I, ST3Gal III and ST3Gal IV in small cell samples. Multiplex PCR amplification using specific primers for each sialyltransferase and detection of amplification products by polyacrylamide gel electrophoresis is a method that is fast and easy to handle and has proven to be useful for establishing sialyltransferase patterns of expression in breast immortalized cell line HBL100 as well as in breast cancer cell lines MCF-7/6, MCF-7/AZ and MDA.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1006983214918
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