Activation of K-ras in transplantable pancreatic ductal adenocarcinomas of Syrian golden hamsters.
Activation of K-ras in transplantable pancreatic ductal adenocarcinomas of Syrian golden hamsters.
复制标题
叙利亚金仓鼠可移植胰腺导管腺癌中 K-ras 的激活。
DOI：
10.1093/carcin/11.11.2075
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发表时间：
1990
期刊：
影响因子：
4.7
通讯作者：
Scarpelli,DG
中科院分区：
文献类型：
--
作者：
Cerny,WL;Mangold,KA;Scarpelli,DG
高摩尔。从正常仓鼠胰腺和两种不同仓鼠可移植癌的固体和腹水变体制备wt基因组DNA，一种由N-亚硝基双(2-氧代丙基)胺诱导，另一种是自发发生的。该DNA被转染到NIH/3T3细胞中，并在注射到裸鼠体内时产生能够形成肿瘤的细胞。通过Southern印迹分析裸鼠肿瘤揭示了仓鼠K-ras特异性条带的存在。由可移植肿瘤制备的基因组 DNA 的 K-rascodon 12-13 区域经聚合酶链式反应 (PCR) 介导扩增，产生 117 by 片段，通过等位基因特异性寡核苷酸杂交和直接 DNA 测序对其进行分析。用特异针对密码子 12 或 13 的第一或第二位置变化的探针进行寡核苷酸杂交，检测到化学诱导的可移植肿瘤中密码子 12 的第二位置的 G 到 A 的颠换，以及自发发生的可移植癌中密码子 13 的第二位置的 G 到 A 的变化。通过对PCR扩增产物进行直接双脱氧测序证实了化学诱导肿瘤的结果。这些发现首次在实验性胰腺肿瘤模型中显示了特定癌基因的激活，并且与最近报道的人类胰腺癌中 K-突变的结果相似。
High mol. wt genomic DNA was prepared from normal hamster pancreas and the solid and ascites variants of two different hamster transplantable carcinomas, one induced byN-nitrosobis(2-oxopropyl)amine and the other spontaneously occurring. This DNA was transfected into NIH/3T3 cells and resulted in cells that were capable of forming tumors when injected into nude mice. Analysis of the nude mouse tumors by Southern blotting revealed the presence of a band specific for hamster K-ras. Polymerase chain reaction (PCR)-mediated amplification of the K-rascodon 12–13 region of genomic DNA prepared from the transplantable tumors produced a 117 by fragment which was analyzed by both allele-specific oligonucleotide hybridization and direct DNA sequencing. Oligonucleotide hybridization with probes specific for changes in the first or second position of codons 12 or 13 detected a G to A transversion in the second position of codon 12 in the chemically induced transplantable tumor, and a G to A change in the second position of codon 13 in the spontaneously occurring transplantable carcinomas. The result obtained for the chemically induced tumor was confirmed by direct dideoxy sequencing of the PCR-amplified product. These findings are the first to show a specific oncogene activation in an experimental pancreatic tumor model and also parallel the results recently reported for K-rasmutations in human pancreatic carcinoma.