Wan D, Zhang X, Zhou X
National Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute.
Zhonghua Yi Xue Za Zhi. 1995 Oct;75(10):606-8, 639.
Using mRNA differential display technique, we studied the interesting cDNA clones of differential expression between human normal liver and hepatocarcinoma. mRNA extracted from normal liver tissue and hepatocarcinoma cell line Hep3B were subjected to RT-PCR reaction with ten combinations of two 3' primers and five 5' primers. After subjecting these PCR products to labelling with 35S(a)-dATP, PAGE and autoradiography, we obtained a lot of cDNA clones of differential expression. Thirty-one of these clones were partially sequenced. It was shown that 30 clones had no sequences matched with GenBank except one. Dot hybridization showed that 2 cDNA clones were overexpressed in liver cancer tissue and 1 cDNA clone was overexpressed in normal liver tissue. The further characterization of these cDNA clones is in progress.
运用mRNA差异显示技术,我们研究了人正常肝脏与肝癌之间差异表达的有趣cDNA克隆。从正常肝组织和肝癌细胞系Hep3B中提取的mRNA,与两种3'引物和五种5'引物的十种组合进行逆转录聚合酶链反应(RT-PCR)。将这些聚合酶链反应产物用35S(α)-dATP标记、进行聚丙烯酰胺凝胶电泳(PAGE)和放射自显影后,我们获得了许多差异表达的cDNA克隆。对其中31个克隆进行了部分测序。结果显示,除一个克隆外,其余30个克隆均无与基因库匹配的序列。斑点杂交表明,2个cDNA克隆在肝癌组织中过度表达,1个cDNA克隆在正常肝组织中过度表达。这些cDNA克隆的进一步鉴定正在进行中。