Differentiation of Pancreatic Endocrine Cells

胰腺内分泌细胞的分化

• 批准号: 08457256
• 负责人: KOJIMA Itaru
• 金额: $ 4.93万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457256/
• 关键词: differentiation; pancreatic endocrine cell; beta-cell; insulin; activin A; betacellulin; hepatocyte growth factor; 膵β細胞; アクチビンA

1) We established a model cell system to study the differentiation of pancreatic endocrine cells.2) Using this sytem, we found that activin A and betacellulin (BTC) convert amylase secreting cells into insulin-secreting cells.3) We also found that hepatocyte growth factor (HGF) reproduce the effect of BTC.4) In AR42J cells, there specific binding sites to BTC.The binding of BTC is replaced by unlabeled BTC but EGF is much less potent. BTC binds to ErbB1 and another protein of MW of 190 KDa, which may be a new member of the EGF receptor family. BTC induced tyrosine phosphorylation of ErbB1, ErbB2, ErbB4 and the 190 KDa protein.5) The differentiation-inducing activity is blocked by an inhibitor of the MAP knase pathway but not by an inhibitor of the Pl 3-kinase pathway. In addition, transfection of Ar42J cells with cDNA for constitutively active MAP kinase kinase induced differentiation. Conversely, transfection of cDNA for MAP inase phosphatase blocked differentiation. Activation of MAP kinase is neccesry and sufficient for the HGF-induced differentiation of AR42J cells.6) We examine the genes expressed during the differentiation of AR42J cells inot insulin-producing cells by differential display. Activin A and BTC induced the expression of 25 genes, 10 of which are unique ones. Expression of some of them was blocked by an inhibitor of MAP kinase. Therefore, these genes are cloosely associated with differentiation of AR42J cells.

1）我们建立了一个模型细胞系统，以研究胰腺内分泌细胞的区分。2）使用该Sytem，我们发现活化蛋白A和贝塔曲霉蛋白（BTC）将淀粉酶分泌的细胞转化为分泌胰岛素分泌细胞的细胞。33）我们还发现肝细胞生长因子（HGF）在btc.42 btc.42j in42J in42j in42J in42J in42j in42j JJ BTC的结合被未标记的BTC取代，但EGF的效力要少得多。 BTC与ERBB1结合，另一种MW的190 kDa蛋白质可能是EGF受体家族的新成员。 BTC诱导的ERBB1，ERBB2，ERBB4和190 kDa蛋白的酪氨酸磷酸化。5）5）分化诱导活性被MAP KNASE途径的抑制剂阻断，而不是被PL 3-激酶途径的抑制剂所阻断。另外，用cDNA转染AR42J细胞用于组成性活性MAP激酶激酶诱导分化。相反，用于地图磷酸酶的cDNA转染封闭了分化。 MAP激酶的激活是必要的，对于HGF诱导的AR42J细胞的分化就足够了。6）我们检查了在AR42J细胞分化过程中表达的基因通过差分显示，因此产生的胰岛素产生细胞。活化素A和BTC诱导了25个基因的表达，其中10个是独特的。其中一些的表达被MAP激酶的抑制剂阻塞。因此，这些基因与AR42J细胞的分化相关。

项目成果

Mashima, H.et al.: "Betacellulin and actirin A coordinately convert amylase-secreting pancreatic AR42J cells int. insulin-secreting cells" Journal of Clinical Investigation. 97. 1647-1654 (1996)

Mashima, H.et al.：“Betacellulin 和 Actirin A 协同将淀粉酶分泌型胰腺 AR42J 细胞转化为胰岛素分泌型细胞”《临床研究杂志》。

Shibata, H., Kanzaki, M., Takeuchi, T., Miyazaki, J.and Kojima, I.: "Two Distinct Signalling Pathways Activated by Activin A in Two Glucose-responsive Pancreatic beta-cell Lines." J.Mol.Endocrinol.16. 249-258 (1998)

Shibata, H.、Kanzaki, M.、Takeuchi, T.、Miyazaki, J. 和 Kojima, I.：“两种葡萄糖反应性胰腺 β 细胞系中激活素 A 激活的两种不同信号通路。”

Ishiyama,N.et al.: "Studies on the betacellulin receptor in AR42J cells." Diabetologia. (印刷中). (1998)

Ishiyama, N. 等人：“AR42J 细胞中 betacellulin 受体的研究”（正在出版）。

Kojima,I.et al.: "Inhibin,Activin and Follistatin" Aono,T.,Sugino,H.,Valle,W., 200 (1997)

Kojima,I.et al.：“抑制素、激活素和卵泡抑素”Aono,T.、Sugino,H.、Valle,W., 200 (1997)

Mashima.H., Ohnishi, H., Wakabayashi, K., Miyagawa, J., Hanafusa, T., Seno, M., Yamada, H.and Kojima, I.: "Betacellulin and Activin A Coordinately Convert Amylase-secreting Pancreatic AR42J Cells into Insulin-secreting Cells." J.Clin.Invest.97. 1647-1654

Mashima.H.、Ohnishi, H.、Wakabayashi, K.、Miyakawa, J.、Hanafusa, T.、Seno, M.、Yamada, H. 和 Kojima, I.：“Betacellulin 和激活素 A 协调转换淀粉酶分泌