Growth inhibition of tumor cells by transfection with β-1,4-glanctosyltransferase gene II or V.

通过转染β-1,4-半乳糖基转移酶基因II或V抑制肿瘤细胞的生长。

• 批准号: 12680708
• 负责人: FURUKAWA Kiyoshi
• 金额: $ 2.3万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680708/
• 关键词: β-1,4-galactosyltransferase; N-glycans; substrate-specificity; gene transfection; tumorgenesis; metastasis; cell growth; cell adhesion; β-1,4-ガラクトース転移酸素; 素質特異性; 遺伝子発現; 癌細胞

Upon malignant transformation of cells, highly branching of N-linked oligosaccharides is induced by activation of N-acetylglucosaminyltransferase V. However, no significant change in β-1, 4-GalT activities was found during this biological event. When gene expression levels of the β-1, 4-GalT family members were examined between malignantly transformed cells and their normal counterparts, those of β-1, 4-GalT II decreased while those of β-1, 4-GalT V increased without significant changes in those of other β-1, 4-GalTs. When B16-F10 mouse melanoma cells were transfected with human β-1, 4-GalT II sense CDNA or β-1, 4-GalT V anti-sense cDNA, and transplanted subcutaneously in C57B/L6 mouse, tumor formation was suppressed in the gene-transfected cells. In order to investigate how tumor suppression was induced by them, the cells were cultured on fibronectin (FN)-coated plates without serum. The mock-transfected B16-F10 cells stayed in a round shape while both gene-transfected cells showed cell spreading aligned with actin filament as revealed with phalloidin staining. Lectin blot analysis of membrane glycoproteins from the tumors and cultured cells using RCA-I which interacts with oligosaccharides terminated with β-1, 4-linked galactose and L-PHA which binds to highly branched oligosaccharides showed that reactivities to these lectins are changed upon transfection of the β-1, 4-GalT genes. These results indicate that the altered galactosylation induced to the membrane glycoproteins including fibronectin receptor affects adhesiveness of the cells to fibronectin substrate, thus leading to the suppression of the tumor formation.

在细胞的恶性转化后，N-乙酰葡萄糖氨基转移酶V诱导了N-连接的寡糖的高度分支检查成员在恶性细胞及其正常的cunterparts eβ-1，4-galt V与其他β-1，4-GALT的显着变化不变。 4-GALT II感应cDNA或β-1，4-GALT V-抗浓度cDNA在C57B/L6小鼠中皮下，在基因转染的细胞中抑制了肿瘤的形成。模拟转染的B16-F10细胞保持圆形，而两个基因转换的细胞与ActIntint的分配均与肿瘤的膜糖蛋白的SIS相结合，并使用RCA-I与RCA-I相互作用。 1，4-连接的半乳​​糖和L-PHA在转化后与这些凝集素结合到这些凝集素的寡糖与膜糖蛋白的转化后变化。因此导致肿瘤形成的抑制。

项目成果

Guo, S.: "Galactosylation of N-linked oligosaccharides by human β-1,4-galactosyl-transferases I, II, III, IV, V and VI expressed in Sf-9 cells"Glycobiology. 11. 813-820 (2001)

郭 S.：“Sf-9 细胞中表达的人 β-1,4-半乳糖基转移酶 I、II、III、IV、V 和 VI 对 N-连接寡糖的半乳糖基化”《糖生物学》11. 813-820 (2001)。 ）

Nakamura, N.: "Differential gene expression of β-1, 4-galactosyltransferases I, II and V during mouse brain development"J. Neurochem.. 76. 29-38 (2001)

Nakamura, N.：“小鼠大脑发育过程中 β-1、4-半乳糖基转移酶 I、II 和 V 的差异基因表达”J. Neurochem.. 76. 29-38 (2001)

Sato, T.: "Correlated gene expression between β-1, 4-galactosyltransferase V and N-acetylglucosaminyltransferase V in human cancer cell lines"Biochem. Biophys. Res. Commun.. 276. 1019-1023 (2000)

Sato, T.：“人类癌细胞系中 β-1, 4-半乳糖基转移酶 V 和 N-乙酰氨基葡萄糖转移酶 V 之间的相关基因表达”Biochem. Biophys Res. 276. 1019-1023 (2000)

Furukawa,K.: "Handbook of Glycosyltransferases and Their Related Genes"Springer-Verlag, Tokyo(in press). (2001)

Furukawa,K.：“糖基转移酶及其相关基因手册”Springer-Verlag，东京（印刷中）。

Sato, T.: "Correlated gene expression between β-1,4-galactosyltransferase V and N-acetylglucosaminyltransferase V in human cancer cell lines"Biochem. Biophys. Res. Commun.. 276. 1019-1023 (2000)

Sato, T.：“人类癌细胞系中 β-1,4-半乳糖基转移酶 V 和 N-乙酰氨基葡萄糖转移酶 V 之间的相关基因表达”Biochem. Biophys Res. 276. 1019-1023 (2000)