Ishihara H, Asano T, Tsukuda K, Katagiri H, Inukai K, Anai M, Kikuchi M, Yazaki Y, Miyazaki J I, Oka Y
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Diabetologia. 1993 Nov;36(11):1139-45. doi: 10.1007/BF00401058.
Glucose-stimulated insulin secretion, glucose transport, glucose phosphorylation and glucose utilization have been characterized in the insulinoma cell line MIN6, which is derived from a transgenic mouse expressing the large T-antigen of SV40 in pancreatic beta cells. Glucose-stimulated insulin secretion occurred progressively from 5 mmol/l glucose, reached the maximal level approximately seven-fold above the basal level at 25 mmol/l, and remained at this level up to 50 mmol/l. Glucose transport was very rapid with the half-maximal uptake of 3-O-methyl-D-glucose being reached within 15 s at 22 degrees C. Glucose phosphorylating activity in the cell homogenate was due mainly to glucokinase; the Vmax value of glucokinase activity was estimated to be 255 +/- 37 nmol.h-1.mg protein-1, constituting approximately 80% of total phosphorylating activity, whereas hexokinase activity constituted less than 20%. MIN6 cells exhibited mainly the high Km component of glucose utilization with a Vmax of 289 +/- 18 nmol.h-1.mg protein-1. Thus, glucose utilization quantitatively and qualitatively reflected glucose phosphorylation in MIN6 cells. In contrast, MIN7 cells, which exhibited only a small increase in insulin secretion in response to glucose, had 4.7-fold greater hexokinase activity than MIN6 cells with a comparable activity of glucokinase. These characteristics of MIN6 cells are very similar to those of isolated islets, indicating that this cell line is an appropriate model for studying the mechanism of glucose-stimulated insulin secretion in pancreatic beta cells.
在胰岛素瘤细胞系MIN6中,已对葡萄糖刺激的胰岛素分泌、葡萄糖转运、葡萄糖磷酸化和葡萄糖利用进行了表征。MIN6细胞系源自一只在胰腺β细胞中表达SV40大T抗原的转基因小鼠。葡萄糖刺激的胰岛素分泌从5 mmol/l葡萄糖时开始逐渐增加,在25 mmol/l时达到比基础水平高约7倍的最大水平,并在50 mmol/l时保持在该水平。葡萄糖转运非常迅速,在22℃下,15秒内即可达到3 - O - 甲基 - D - 葡萄糖摄取的半最大值。细胞匀浆中的葡萄糖磷酸化活性主要归因于葡萄糖激酶;葡萄糖激酶活性的Vmax值估计为255±37 nmol·h⁻¹·mg蛋白⁻¹,约占总磷酸化活性的80%,而己糖激酶活性占比不到20%。MIN6细胞主要表现出葡萄糖利用的高Km组分,Vmax为289±18 nmol·h⁻¹·mg蛋白⁻¹。因此,葡萄糖利用在数量和质量上反映了MIN6细胞中的葡萄糖磷酸化。相比之下,MIN7细胞对葡萄糖的胰岛素分泌仅略有增加,其己糖激酶活性比具有相当葡萄糖激酶活性的MIN6细胞高4.7倍。MIN6细胞的这些特征与分离的胰岛非常相似,表明该细胞系是研究胰腺β细胞中葡萄糖刺激的胰岛素分泌机制的合适模型。