How dose the gastric surface mucus cell line derived from transgenic mouse synthesize and secrete mucins?

转基因小鼠胃表面粘液细胞系如何合成和分泌粘蛋白？

• 批准号: 07680773
• 负责人: HOTTA Kyoko
• 金额: $ 1.41万
• 依托单位: KITASATO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680773/
• 关键词: Hyaluronan; Gastric mucosa; Gastric epithelial cells; Transgenic mouse; SV40; large T-antigen; GSM06 cells; ムチン; 粘液細胞; 胃粘液; モノクロナール抗体; 胃表層粘液細胞

GSM06 is a cell line established from the stomach of transgenic mouse harboring a temperature-sensitive simian virus 40 (SV40) large T-antigen gene. ^3H-Labeled macromolecules produced by the cells incubated with [^3H] glucosamine were characterized to examine whether or not GSM06 cells synthesize mucin (mucus glycoprotein). The GSM06 cells grew until a confluent monolayr formed at 33ﾟC (the permissive temperature for SV40 large T-antigen expression), and the ^3H-labeled macromolecules appeared in both cell extract and medium during culture for at least one week. Unexpectedly, almost all ^3H-labeled macromolecules, which were excluded from a column of Sepharose CL-4B,were identified as hyaluronan by analyzes using Sepharose CL-2B chromatography, cesium trifluoroacetate equilibrium centrigugation, treatment with dithiothreitol, and trypsin and hyaluronidase digestion. At a nonpermissive temperature (39ﾟC), GSM06 cells grew only slightly, but produced much more hyaluronan than at 33ﾟC.The results indicate that GSM06 cells do not produce mucin, but hyaluronan, and that the expression of large T-antigen may influence hyaluronan synthesis in GSM06 cells.

GSM 06是从携带温度敏感性猿猴病毒40（SV 40）大T抗原基因的转基因小鼠胃中建立的细胞系。用[^3H]葡糖胺孵育细胞产生的^3H标记的大分子被表征，以检查GSM 06细胞是否合成粘蛋白（粘液糖蛋白）。在33 ℃（SV 40大T抗原表达的允许温度）下，GSM 06细胞生长直至形成融合单层，在至少一周的培养过程中，细胞提取物和培养基中都出现了^3H标记的大分子。出乎意料的是，通过琼脂糖CL-2B层析、三氟乙酸铯平衡离心、二硫苏糖醇处理、胰蛋白酶和透明质酸酶消化，几乎所有从琼脂糖CL-4 B柱中排除的^3H标记的大分子都被鉴定为透明质酸。在非允许温度（39 ℃）下，GSM 06细胞仅轻微生长，但产生比33 ℃多得多的透明质酸。结果表明，GSM 06细胞不产生粘蛋白，但产生透明质酸，并且大T抗原的表达可能影响GSM 06细胞中透明质酸的合成。

项目成果

Watanabe T.他4名: "Mechanisms for cytoprotection by vitamin U from ethanol-induced gastric mucosal damage in rats" Dig.Dis.Sci.41. 49-54 (1996)

Watanabe T. 和其他 4 人：“维生素 U 对大鼠乙醇诱导的胃粘膜损伤的细胞保护机制”Dig.Dis.Sci.49-54 (1996)。

Kazuhiko Ishihara et al.: "Establishment of monoclonal antibodies against carbohydrate moiety of gastric mucins distributed in the different sites and layrs of rat gastric mucosa" Glycoconj.J.13. 857-864 (1996)

Kazuhiko Ishihara 等人：“针对分布在大鼠胃粘膜不同部位和层的胃粘蛋白碳水化合物部分的单克隆抗体的建立”Glycoconj.J.13。

K. Ishihara他4名: "A Monoclonal Antibody to the Gastric Gland Mucous Cell-Derived Mucin, HIK-1083, Reacts with Oligosaccharides Bearing Peripheral α-Linked GlcNAc Residues" Glycoconj. J.12. 504- (1995)

K. Ishihara 和其他 4 人：“胃腺粘液细胞衍生粘蛋白的单克隆抗体，HIK-1083，与带有外周 α-连接 GlcNAc 残基的寡糖发生反应”Glycoconj 504- (1995)。

Yukinobu Goso他3名: "Effects of traditional herbal medicine on gastric mucin against ethanol-induced gastric injury in rats" Comp.Biochem.Phsiol.C. 113. 17-21 (1996)

Yukinobu Goso 等 3 人：“传统草药对胃粘蛋白对大鼠乙醇诱导胃损伤的影响”Comp.Biochem.Phsiol.C. 113. 17-21 (1996)

Kazuhiko Ishihara他6名: "Estasblishment of monoclonal antibodies against carbohydrate moiety of gastric mucins distributed in the different sites and layers of rat gastric mucosa" Glycoconj.J.13. 857-864 (1996)

Kazuhiko Ishihara 等 6 人：“针对分布在大鼠胃粘膜不同部位和层的胃粘蛋白碳水化合物部分的单克隆抗体的建立”Glycoconj.J.13（1996）。