Gold G, Gishizky M L, Chick W L, Grodsky G M
Diabetes. 1984 Jun;33(6):556-61. doi: 10.2337/diab.33.6.556.
A series of 3H-leucine pulse-labeling experiments was carried out with dispersed cells freshly isolated from transplanted rat insulinomas. After secreted fractions were separated, insulin was purified and specific activities were determined for both secreted and average cellular insulins. Labeling patterns in this line of tumor cells were compared with those previously established for isolated rat islets. With both tumors and islets, conversion of labeled proinsulin to insulin occurred to the same extent by 2.5 h, suggesting similar onset and half-time of proteolysis in these cells. However, total cellular insulin in tumors attained a threefold higher specific activity than in islets. Because total B-cell mass in these tumors was unknown, either a more rapid proinsulin biosynthesis or diminished cellular storage (or both) could lead to this faster fractional replacement of total stored insulin. Insulin secretion in these tumor cells was insensitive to high glucose but responded, albeit poorly, to leucine plus 3-isobutyl-1-methylxanthine (IBMX). Under all secretory conditions tested, tumor cells continuously secreted insulin at elevated fractional rates, which were slightly higher than fractional insulin secretory rates in maximally glucose-stimulated islets. In contrast with normal islets, newly synthesized insulin was stored homogeneously in tumor cells, and compartmental storage characteristics were not generated by incubation with either 20 mM glucose or leucine plus IBMX in the marking period. Thus, preferential secretion of insulin was never observed in tumor cells.(ABSTRACT TRUNCATED AT 250 WORDS)
对从移植的大鼠胰岛素瘤中新鲜分离的分散细胞进行了一系列3H-亮氨酸脉冲标记实验。分离分泌部分后,纯化胰岛素并测定分泌胰岛素和平均细胞胰岛素的比活性。将该肿瘤细胞系中的标记模式与先前为分离的大鼠胰岛建立的模式进行比较。在肿瘤和胰岛中,标记的胰岛素原在2.5小时内转化为胰岛素的程度相同,表明这些细胞中蛋白水解的起始和半衰期相似。然而,肿瘤中的总细胞胰岛素比胰岛中的比活性高三倍。由于这些肿瘤中的总B细胞量未知,胰岛素原生物合成更快或细胞储存减少(或两者兼有)都可能导致总储存胰岛素的这种更快的部分替代。这些肿瘤细胞中的胰岛素分泌对高葡萄糖不敏感,但对亮氨酸加3-异丁基-1-甲基黄嘌呤(IBMX)有反应,尽管反应较差。在所有测试的分泌条件下,肿瘤细胞以升高的分数率持续分泌胰岛素,略高于最大葡萄糖刺激的胰岛中的分数胰岛素分泌率。与正常胰岛相反,新合成的胰岛素在肿瘤细胞中均匀储存,并且在标记期与20 mM葡萄糖或亮氨酸加IBMX孵育不会产生区室化储存特征。因此,在肿瘤细胞中从未观察到胰岛素的优先分泌。(摘要截断于250字)