Pochet R, Blachier F, Lawson D E, Malaisse W J
Laboratory of Histology, Faculté de Médecine, Université Libre de Bruxelles, Belgium.
Int J Pancreatol. 1989 Oct;5(3):295-304. doi: 10.1007/BF02924474.
Calbindin-D 28K expression in insulin-producing tumoral cells of the RINm5F line was assessed by Western-blot and high pressure liquid chromatography. Western blot analysis demonstrated the presence in RINm5F cell homogenates of a protein recognized by a specific polyclonal antibody against chick calbindin. Proteins with apparent molecular weights (mol wt) of 44, 47, 56, and 85 kD were also recognized by the antiserum in RINm5F cell extract, but not in normal rat islet extract. HPLC heat-resistant protein extract from RINm5F cell homogenates revealed three calbindin positive peaks: a major peak with a retention time (20.5 min) identical to that found in a rat cerebellar extract and two minor peaks with shorter retention times. The calbindin content of RINm5F cells was apparently unaffected after 9 d culture in a medium supplemented with 10% calf serum pretreated with dextran-charcoal to remove 1,25-dihydroxyvitamin D3.
通过蛋白质免疫印迹法和高压液相色谱法评估了RINm5F细胞系中产生胰岛素的肿瘤细胞中钙结合蛋白-D 28K的表达。蛋白质免疫印迹分析表明，在RINm5F细胞匀浆中存在一种可被抗鸡钙结合蛋白的特异性多克隆抗体识别的蛋白质。抗血清在RINm5F细胞提取物中还识别出表观分子量（mol wt）为44、47、56和85 kD的蛋白质，但在正常大鼠胰岛提取物中未识别出。来自RINm5F细胞匀浆的HPLC耐热蛋白提取物显示出三个钙结合蛋白阳性峰：一个主峰的保留时间（20.5分钟）与大鼠小脑提取物中的相同，还有两个保留时间较短的小峰。在补充了用葡聚糖-活性炭预处理以去除1,25-二羟基维生素D3的10%小牛血清的培养基中培养9天后，RINm5F细胞的钙结合蛋白含量显然未受影响。
