Parsa I, Cleary C M, Marsh W H, Butt K M, Foye C A
State University of New York, Health Science Center, Brooklyn.
Int J Pancreatol. 1986 Dec;1(5-6):299-308. doi: 10.1007/BF02801863.
c-Ki-ras-2 sequences were visualized in paraffin-embedded sections from normal fetal and adult human pancreases, a chemically induced transplantable human pancreas carcinoma (PT-1) and three carcinomas of pancreas by in situ hybridization technique. A biotinylated 1-kilobase-pair (kb) EcoRI fragment of pHiHi3 DNA was used as probe and the oncogene was visualized as one or two large grains of reaction products produced by streptavidin-peroxidase complex and diaminobenzidine tetrachloride in more than 9% of normal pancreas nuclei. Its amplification in the chemically induced cell line was detected as one or more large grains in 72% of the nuclei and numerous cytoplasmic grains. The detection of oncogene in normal pancreases and its amplification in PT-1 cells was validated by Southern analysis of EcoRI digests of genomic DNA extracted from normal pancreases and PT-1 cell line. The oncogene was also demonstrated to be equally amplified in two adenocarcinomas and one undifferentiated carcinoma of human pancreas by in situ hybridization.
通过原位杂交技术,在正常胎儿和成人胰腺、化学诱导的可移植性人胰腺癌(PT-1)以及三种胰腺癌的石蜡包埋切片中观察c-Ki-ras-2序列。用生物素化的pHiHi3 DNA的1千碱基对(kb)EcoRI片段作为探针,在超过9%的正常胰腺细胞核中,癌基因表现为链霉亲和素-过氧化物酶复合物和四氯化二氨基联苯胺产生的一或两个大的反应产物颗粒。在化学诱导的细胞系中,其扩增表现为72%的细胞核中有一或多个大颗粒以及众多细胞质颗粒。从正常胰腺和PT-1细胞系提取的基因组DNA经EcoRI消化后的Southern分析证实了正常胰腺中癌基因的存在及其在PT-1细胞中的扩增。原位杂交还显示该癌基因在人胰腺的两个腺癌和一个未分化癌中同样发生扩增。