应用mRNA差异显示技术克隆新的肝癌相关基因的研究

Study on cloning of novel liver cancer associated genes using mRNA differential display method

目的 研究和克隆新的肝癌相关基因 ,探讨肝癌发生的分子学基础。方法 采用mRNA差异显示技术 (DDPCR)和逆转录聚合酶链式反应、测序等方法 ,分析 2对肝细胞癌和癌旁组织基因表达差异情况 ,并用Northern印迹杂交的方法 ,分析所获得的基因片段在 12对肝细胞癌和癌旁组织中的表达。结果 使用 5条上游引物、4条下游引物 ,从 2对肝癌和癌旁组织中获得 6个新的肝癌组织高表达基因片段STW 1、STW 2、STW 3、STW 4、STW 5和STW 6 ,其在肝癌组织中表达的阳性率分别是 :83 .3% (10 /12 )、6 6 .7% (8/12 )、5 0 .0 % (6 /12 )、83 .3 % (10 /12 )、5 8.3% (7/12 )和 6 6 .7%(8/12 ) ,且所分析的 12对肝癌组织都存在 2种以上的新基因片段表达。结论 用DDPCR技术获得6个新的肝癌差异表达基因片段 ,说明肝癌的发生、发展是多基因参与的复杂过程。

Objective\ To study and clone novel liver cancer related genes, and to explore the molecular basis of liver cancer genesis. Methods\ Using mRNA differential display polymerase chain reaction (DDPCR), reverse transcription and ploymerase chain reaction(RT-PCR) and sequencing, we investigated the difference of mRNA of two pairs of specimen, and analyzed the expression of these novel genes in 12 pair of human hepatocellular carcinoma and paracarcinoma tissue by means of Northern Blot. Results\ By 5 upstream primers and 4 downstream primers, we have got 6 novel genes high expressed in liver cancer from two pairs of specimen, named STW-1, STW-2, STW-3, STW-4, STW-5 and STW-6. The positive expression rate of these genes was 83.3%(10/12), 66.7%(8/12), 50.0%(6/12), 83.3%(10/12), 58.3% (7/12) and 66.7%(8/12) separately. There existed more than two kinds of genes expressed in these 12 pair of samples. Conclusion\ We have got 6 novel differential expressing genes using DDPCR method, and indicating that is a complicated process of many oncogenes and antioncogenes participating in hepatocellular carcinoma genesis.\;