| Objective:To examine the membraneα2,3 polysialic acid residues of AGS, a human gastric cancer cell line,and estimate their effects on the growth and migration ability of AGS, and investigate the expression of severalα2,3 linked polysialic acids related genes' mRNA-level expression.Methods:(1)The different neucleotide sequences of theα2,3 sialyltransferases family were obtained based on their respective accession numbers, and six pairs of respective primers were designed by software based on their different neucleotide sequences. So RT-PCR was carried out to determine the expression of the six members at the mRNA level in gastric cell AGS.(2)The growth of AGS cells was evaluated through MTT, based on various conditions ofα2,3-Neuraminidase incubation, including different enzyme concentrations.(3)The migration ability of AGS cells was evaluated through wound-healing assay, based on various conditions ofα2,3-Neuraminidase incubation,including duration and enzyme concentration.(4)AGS cell surface average fluorescence intensity was investigated with FCM through fluorescence-labeled Maackia amurensis lectin I (MAL I) ,which is highly specific withα2,3-polysialic acid residues, based on various conditions ofα2,3-Neuraminidase incubation, including enzyme concentration.Results:As the RT-PCR data showed, in gastric cell AGS, at least two members of theα2,3 sialyltransferases family,namely, ST3Gal and和ST3Gal4,were expressed at the mRNA level. As indicated by the growth curve of AGS, the catalytic and inhibitive ability ofα2,3 sialidase was best exhibited when the enzyme concentration ranges from 100mU/ml,150mU/ml to 500mU/ml,and the incubation time lasted for four days or 96 hours,and the growth cycle of AGS significantly decreased compared with the control group. Meanwhile, the wound healing assay proved that the membraneα2,3-polysialic acid residues played a role in the migration capacity of AGS, because theα2,3 sialidase treatment significantly increased the migration ability of AGS.Conclusions: A least two members of theα2,3 sialyltransferases family were expressed in gastric cell AGS. And the membraneα2,3-polysialic acid residues is related to the growth of AGS, supporting its growth to some extent and shortening its growth cycle. Also the membraneα2,3-polysialic acid residues of AGS cells has a negative impact on the migration ability of AGS, indicating that the membraneα2,3-polysialic acid residues can play a role in cell-cell signal transduction and thereby modifying the biological behaviour of gastric cell AGS.
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