摘要
以5例原发性肝癌组织的pay(A^+)RNA构建一个1×10~6的混合肝癌cDNA文库,并以正常人肝IGF-Ⅱ基因片段为探针筛选此文库,得到多个阳性克隆,从中再以一个较长片段的克隆作为探针对其它组织作Northern杂交分析,发现胎脯、胎肝及其它肝癌组织中的IGF-Ⅱ基因都有过量表达,而正常成年肝组织无明显表达。
To investigate the roles played by oncogenes in the devdopment and progression of human primaty hepatic cancer (PHC), a liver cancer cDNA library was constructed, and mRNA was isolated from 5 samples of human PHC, and was purified by chromatography on oligo (dT12-18) primer, and it was ligased to BstXl adaptor and cloned into plasmid pCDM8 which can be expressed in eukaryotic cells and analyzed in bacteria, and then transformed into bacteria MC1061/P3. Finally about 1×10~6/5μg mRNA cDNA library was constructed.
An IGF-Ⅱ gene was used as a probe to screen the library. After 3 rounds of screening, 5 positive clones were obtained, the largest clone of which was chosen to be used as a probe in Northern blot analysis. Results showed that this gene was overexpressed in fetal liver, fetal brain and liver cancer tissue vs that in the normal adult human liver. This suggested that the gone may be related to liver cancer and fetal growth.
出处
《上海第二医科大学学报》
CSCD
1993年第4期309-313,共5页
Acta Universitatis Medicinalis Secondae Shanghai
