Immortalized bovine pancreatic duct cells become tumorigenic after transfection with mutant k-ras
Immortalized bovine pancreatic duct cells become tumorigenic after transfection with mutant k-ras
复制标题
DOI：
10.1007/s004280100397
复制
发表时间：
2001-06-01
期刊：
影响因子：
3.5
通讯作者：
Jesnowski, R
中科院分区：
文献类型：
--
作者：
Löhr, M;Müller, P;Jesnowski, R
K-ras基因突变被认为是胰腺癌发生的早期和重要事件。为了研究这种分子变化在正常胰腺细胞向肿瘤细胞转变过程中的作用，首先用SV40大T抗原(Ag)互补(C)DNA转染牛胰管细胞，然后再将突变的K-ras基因导入牛胰管细胞。与原代导管细胞一样，永生化的导管细胞(超过100代)表达细胞角蛋白、碳酸氢酶-II、囊性纤维化跨膜传导调节因子(CFTR)和多药耐药(MDR)。移植到塑料穹顶下后，它们以单层的形式生长，在Matrigel上生长时形成了类似于管道的三维结构。胰岛素、表皮生长因子(EGF)、转化生长因子(TGF)-α刺激细胞生长，胃泌素、CCK-8对细胞无反应。它们不会在软琼脂中形成克隆，也不会在裸鼠体内形成肿瘤。K-ras基因突变的永生化细胞在裸鼠胰腺内原位注射后获得成瘤能力。结论：SV-40永生化牛胰管细胞通过转导突变的K-ras基因获得了致瘤性，并保留了正常胰管细胞的特性。这表明K-ras在胰腺癌模型中发挥了重要作用。
Mutation of the K-ras gene is thought to be an early and important event in pancreatic carcinogenesis. In order to study the role of this molecular alteration in the transition from the normal to the neoplastic pancreatic cell, bovine pancreatic duct cells were first immortalized by SV40 large T antigen (Ag) complementary (c)DNA transfection and then transfected with a mutated K-ras gene. As did primary duct cells, the immortalized duct cells (more than 100 passages) expressed cytokeratins, carbonic anhydrase type-II, cystic fibrosis transmembrane conductance regulator (CFTR), and multidrug resistance (mdr). They grew as a single layer after transplantation under plastic domes and formed three-dimensional structures resembling ducts when grown on Matrigel. Cell growth was stimulated by insulin, epidermal growth factor (EGF), transforming growth factor (TGF)-alpha, but cells did not respond to gastrin and CCK-8. They did not form colonies in soft agar nor did they form tumors in nude mice. Immortalized cells transfected with mutated K-ras acquired the ability to form tumors after orthotopic injection into the nude mouse pancreas. It is concluded that SV 40 immortalized bovine pancreatic duct cells retain the features of normal duct cells and gain tumorigenicity by transfection with mutated K-ras. This suggests an important role for K-ras in this pancreatic carcinoma model.