正常肝与肝癌细胞表达差异的cDNA克隆的研究

采用信使核糖核酸（ｍＲＮＡ）差异显示技术（ｍＲＮＡＤｉｆｆｒｅｎｔｉａｌＤｉｓｐｌａｙｍＲＮＡＤＤ），寻找人正常肝与肝癌细胞表达差异的互补脱氧核糖核酸（ｃＤＮＡ）克隆。人正常肝组织及人肝癌细胞株（Ｈｅｐ３Ｂ）ｍＲＮＡ用３＇端二个引物分别与五种不同５＇端引物进行逆转录─聚合酶链反应（ＲＴ－ＰＣＲ），以３５Ｓ－α（ｄＡＴＰ）参入产物进行聚丙烯酰胺凝胶电泳（ＰＡＧＥ）及放射自显影。获得一批表达差异的ｃＤＮＡ克隆。选择其中３１个克隆进行部分ＤＮＡ序列分析，除一个为已知序列外其余３０个克隆为ＧｅｎｅＢａｎｋ中无匹配序列。经斑点杂交证明二个克隆在肝癌中高表达，一个克隆在正常肝中高表达。目前正在对有意义的克隆做进一步论证。

cDNA clones of differential expression between human normal liver tissue and hepatocarcinoma

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 RTPCR 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.