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)我们建立了研究胰腺内分泌细胞分化的模型细胞系统。2)利用该系统，我们发现激活素A和β细胞蛋白(BTC)可以将淀粉酶分泌细胞转化为胰岛素分泌细胞。3)我们还发现肝细胞生长因子(HGF)在AR42J细胞中复制了BTC.4的作用，存在与BTC的特异结合位点。BTC的结合被未标记的BTC取代，而EGF的结合作用要弱得多。BTC与ErbB1和另一个相对分子质量为190 KDa的蛋白结合，可能是EGF受体家族的新成员。BTC诱导ErbB1、ErbB2、ErbB4和190 KDa蛋白的酪氨酸磷酸化。5)分化诱导活性可被MAP-Kase通路的抑制剂阻断，但不能被P13-K通路的抑制剂阻断。此外，将具有结构性活性的MAPKK基因导入Ar42J细胞可诱导分化。反之，MAP酶磷酸酶基因的表达抑制了细胞的分化。MAPK的激活是HGF诱导AR42J细胞分化的必要条件和充分条件。6)利用差异显示技术检测AR42J细胞在非胰岛素分泌细胞分化过程中表达的基因。激活素A和BTC诱导了25个基因的表达，其中10个是独一无二的。其中一些基因的表达被一种MAP激酶抑制剂阻断。因此，这些基因与AR42J细胞的分化密切相关。

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