Analysis of Antitumor Glycopeptide Containing Sialic Acid At High Level Obtained From Hen Egg White

从鸡蛋清中提取的高含量唾液酸抗肿瘤糖肽的分析

• 批准号: 03806042
• 负责人: WATANABE Kenji
• 金额: $ 1.28万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03806042/
• 关键词: Hen Egg Thick White Protein; beta-Ovomucin; Antitumor Activity; Sialoglycoprotein; 鶏卵濃厚卵白; βーオボムチン; 鶏卵農厚卵白

Glycoprotein (HSGT.molecular weight :400kDa) containing sialic acid at high level was fractionated by the general method from ovomucin in the thick egg white of fresh hen egg. and then sialoglycopeptide (HSGPE) was also fractionated by the methods of gel-filtration and affinity chromatography from the HSGT digested with trypsin. When the tumor cells such as Melanoma cell. Lewis lung cancer cell. Heptoma cell. SR180 cell and the like were incubated with the prepared samples. the clear decrease or evanescence of microvilli and the definitive swelling like petal of microvilli were observed with scanning electron microscope. with the increasings of the added amounts of samples and the incubation time. Moreover. when the described-above tumor cells were also incubated with the same prepared samples in the presence of 6-^3H-tymidine to determine their inhibitory activity for the synthesis of radioactive nucleotides. the decreasings of DNA synthesis in their cells were observed at the decreased levels of 15-40%. comparing with the case of no addition of samplces. The decreased ratios were different every tumor cells. These results indicated that the sensibility of the cells against samples was different. On the other hand. the morphological change in the normal cells incubated with the prepared samples was not observed. Thus. HSGT and HSGPE were found to have antitumor activities for the tumor cells.

采用常规方法从新鲜蛋清中分离出高含量唾液酸的糖蛋白(HSGT，分子量为400 kDa)。然后用凝胶过滤和亲和层析的方法从胰酶消化的HSGT中分离得到唾液糖肽(HSGPE)。当肿瘤细胞如黑色素瘤细胞。Lewis肺癌细胞。七人瘤细胞。将制备好的样品与SR180细胞等孵育。扫描电子显微镜下可见微绒毛明显减少或消失，微绒毛明显肿胀，呈花瓣状。随着样品加入量的增加和孵化时间的延长。更有甚者。将上述肿瘤细胞与相同制备的样品在6-~1H-胸腺嘧啶核苷存在下孵育，测定其对放射性核苷酸合成的抑制活性。其细胞DNA合成下降幅度在15-40%之间。与不添加样品的情况相比。不同肿瘤细胞的减少率不同。这些结果表明，细胞对样品的敏感性是不同的。从另一方面来说。与制备的样品孵育的正常细胞未观察到形态变化。因此。HSGT和HSGPE对肿瘤细胞具有抗肿瘤活性。

项目成果

渡邊乾二: "乳の感染防御因子" 乳技協資料. 42. 1-14 (1992)

Kenji Watanabe：“牛奶感染防御因子”牛奶技术协会资料 42. 1-14 (1992)。

大網 弘: "鶏卵卵白オボムチン由来シアロ糖蛋白質の腫瘍細胞傷害作用" 医学と生物学. 126. 19-23 (1993)

Hiroshi Oami：“源自鸡蛋清卵粘蛋白的唾液酸糖蛋白的肿瘤细胞毒性作用”医学与生物学，126. 19-23 (1993)。

中村 良,川岸 舜朗: "食品分析学" 文永堂出版, 285 (1991)

Ryo Nakamura、Shunro Kawagishi：“食品分析” Buneido Publishing，285（1991）

細野 明義: "ミルクの新しい働き" 全国農協乳業プラント協会, 130 (1991)

细野昭吉：《牛奶的新功能》全国农业合作社乳制品工厂协会，130（1991）

大網弘: "鶏卵卵白オボムチン由来シアロ糖蛋白質の腫瘍細胞傷害作用" 医学と生物学. 126. 19-23 (1993)

Hiroshi Oami：“源自鸡蛋清卵粘蛋白的唾液酸糖蛋白的肿瘤细胞毒性作用”医学与生物学，126. 19-23 (1993)。