Analysis of cellular functions of cells transfected with sense or antisense β-1,4-galactosyltransferase cDNA

正义或反义β-1,4-半乳糖基转移酶cDNA转染细胞的细胞功能分析

• 批准号: 10680696
• 负责人: FURUKAWA Kiyoshi
• 金额: $ 2.11万
• 依托单位: Tokyo Metropolitan Institute of Gerontology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680696/
• 关键词: β-1,4-galactosyltransferase; mouse brain; site-specific distribution; tumor cells; neural cells; sense cDNA; antisense cDNA; altered properties; 組織特異的発現; 突起形成

Mammalian cells are now shown to contain several β-1、4-galactosyltransferases (β-1,4-GalTs) which are involved in the biosynthesis of glycoprotein. When cells are transformed, they express the highly branched sugar chains with tumor-associated carbohydrate antigens on proteins which are critical to tumorigenicity and metastasis of the cell. During transformation, the gene expression levels of β-1,4-GalTs I, III, IV and VI were not changed while that of β-1,4-GalT II decreased and that of β-1,4-GalT V increased. In order to investigate biological significance of the carbohydrates formed by β-1,4-GalT II and V, their genes were introduced into several cell lines. When β-1,4-GalT II-sense cDNA or β-1,4-GalT V-antisense cDNA was transfected into mouse melanoma F10 cells, altered glycosylation of the membrane glycoproteins was detected by lectin blot analysis. The cells were inoculated into C57B mouse and the ability to form tumors were examined. Mouse melanoma F10 cells formed tumors in the animals while those transfected with β-1,4-Galt II-sense cDNA or β-1,4-GalT V-antisense cDNA failed or developed tumors very slowly. These results indicated that the sugar chains synthesized by β-1,4-GalTs II and V are involved in the growth control mechanism of cells. Further studies are necessary to identify which carbohydrate structures are really engaged in this biological event.

现在显示哺乳动物细胞含有几种参与糖蛋白生物合成的β-1,4-半乳糖基转移酶（β-1,4-GalT）。当细胞转化时，它们表达具有肿瘤相关碳水化合物的高度支化糖链。在转化过程中，β-1,4-GalTs I、III、IV 和 VI 的基因表达水平没有改变。 β-1,4-GalT II 的基因减少，β-1,4-GalT V 的基因增加。为了研究β-1,4-GalT II 和V 形成的碳水化合物的生物学意义，对它们的基因进行了研究。当β-1,4-GalT II-sense cDNA或β-1,4-GalT V-antisense cDNA转染到小鼠黑色素瘤F10细胞中时，膜的糖基化发生改变。将细胞接种到C57B小鼠体内，并检测转染β-1,4-Galt II-sense cDNA或β的小鼠黑色素瘤F10细胞在动物体内形成肿瘤的能力。 -1,4-GalT V-反义 cDNA 失败或肿瘤生长非常缓慢。这些结果表明 β-1,4-GalTs II 和 V 合成的糖链是需要进一步研究以确定哪些碳水化合物结构真正参与了这一生物事件。

项目成果

K. Shirane: "Involvement of β-1, 4-galactosyltransferase Vin malignant transformation-associated changes in glycosylation"Biochem. Biophys. Res. Commun.. 265. 434-438 (1999)

K. Shirane：“糖基化中β-1, 4-半乳糖基转移酶 Vin 恶性转化相关变化的参与”Biochem. Commun. 265. 434-438 (1999)

Sato, T.: "Involvement of recombinant human β-1,4-galactosyltransferase V in lactosylceramide biosynthesis."Res. Commun. Biochem. Cell & Molec. Bio.. (in press).

Sato, T.：“重组人 β-1,4-半乳糖基转移酶 V 参与乳糖神经酰胺生物合成”。Res. Commun. Cell & Molec。

T.Sato,et al.(3): "Differential effect of α-lactalbumin on β-1,4-galactosyltransferase IV activities" Biochem.Biophys.Res.Commun.244. 637-641 (1998)

T. Sato 等人 (3)：“α-乳清蛋白对 β-1,4-半乳糖基转移酶 IV 活性的不同影响”Biochem.Biophys.Res.Commun.244 (1998)。

Kido, M.: "Normal levels of serum glycoproteins maintained in β-1,4-galactosyltransferase I-knockout mouse."FEBS Lett.. 464. 75-79 (1999)

Kido, M.：“β-1,4-半乳糖基转移酶 I 敲除小鼠中血清糖蛋白维持正常水平。”FEBS Lett.. 464. 75-79 (1999)

Shirane, K.: "Involvement of β-1,4-galactosyltransferase V in malignant transformation associated changes in glycosylation."Biochem. Biophys. Res. Commun.. 265. 434-438 (1999)

Shirane, K.：“β-1,4-半乳糖基转移酶 V 参与恶性转化相关的糖基化变化。”Biochem. Biophys Commun. 265. 434-438 (1999)