Over-expression of beta-1,4-galactosyltransferase V increases the growth of astrocytoma cell line.
Over-expression of beta-1,4-galactosyltransferase V increases the growth of astrocytoma cell line.
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发表时间：
2002-09
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通讯作者：
S. Xu;S. Zhang;C. Chen;J. Yan;M. Cai;X. Zhu;J. Gu
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作者：
S. Xu;S. Zhang;C. Chen;J. Yan;M. Cai;X. Zhu;J. Gu
我们的前期研究表明，在星形细胞瘤的进展过程中，β-1，4-半乳糖基转移酶V（beta-1，4-GalT V）基因表达增加，优先半乳糖基化寡糖的GlcNAc 1-> 6 Man，在星形细胞瘤IV级中表达最高。为了研究β-1，4-GalT在细胞增殖中的作用，将β-1，4-GalT V的正义和反义cDNA分别构建成pcDNA 3-HA-GalT V和pcDNA 3-anti-GalT V，并转染人星形细胞瘤细胞系SHG。用北方和Western印迹法证实转染。结果表明，SHG/HA-GalTV-AS在含血清培养基中的生长速度明显快于pcDNA 3的模拟转染细胞，而SHG/GalTV-AS的生长速度则明显慢于pcDNA 3的模拟转染细胞。软琼脂贴壁生长实验表明，GalTV-HA/SHG比GalTV-AS/SHG具有更强的集落形成能力。这一结果与生长曲线一致。经RCA-1凝集素染色，GalTV-HA/SHG和SHG/GalTV-AS细胞表面的半乳糖基化程度分别较对照组增强（P<0.001）和减弱（P<0.05）。这表明β-1，4-GalT V参与星形细胞瘤细胞的恶性表型，可能导致细胞表面的高半乳糖基化。
Our previous study showed that the gene expression of beta-1,4-galactosyltransferase V (beta-1,4-GalT V), preferentially galactosylating GlcNAc1-->6Man of oligosaccharides, increased in the process of astrocytoma progress, with the highest level in grade IV astrocytoma. To investigate the function of this beta-1,4-GalT in cell proliferation, the sense and antisense cDNA of beta-1,4-GalT V was constructed as pcDNA3-HA-GalT V and pcDNA3-anti-GalT V respectively and transfected into SHG cell, a kind of human astrocytoma cell line. The transfection was confirmed with Northern and Western blot assay. It was found that the growth of SHG/HA-GalT V in serum-containing medium was faster than that of mock-transfectant with the vector pcDNA3, whereas the growth of SHG/GalTV-AS was slower than that of mock-transfectant. GalTV-HA/SHG showed a stronger capability for colony formation than that of GalTV-AS/SHG as evaluated by anchorage-independent growth in soft agar assay. This result was consistent with that of the growth curve. By RCA-1 lectin assay, the galactosylation on the surface of GalTV-HA/SHG and SHG/GalTV-AS was stained stronger (P<0.001) and weaker (P<0.05) respectively compared with the mock transfectant. This indicates that beta-1,4-GalT V was involved in the malignant phenotype of astrocytoma cells, possibly causing the high galactosylation on the cell surface.