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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 new liver cancer related genes and explore the molecular basis of liver cancer. Methods Differential gene expression technology (DDPCR), reverse transcription polymerase chain reaction and sequencing were used to analyze the gene expression differences of two pairs of hepatocellular carcinoma and paracancerous tissues. The obtained gene fragments were analyzed by Northern blot hybridization. Expression in 12 pairs of hepatocellular carcinoma and paracancerous tissues. Results Using five upstream primers and four downstream primers, six new liver cancer tissue high expression gene fragments STW 1, STW 2, STW 3, STW 4, STW 5 and STW 6 were obtained from 2 pairs of liver cancer and adjacent tissues. The positive rates of HCC expression in hepatocellular carcinoma were: 83.3% (10/12), 66.7% (8/12), 50% (6/12), 83.3% (10). /12), 5 8.3% (7/12) and 66.7% (8/12), and there were more than two novel gene fragments in 12 pairs of liver cancer tissues analyzed. Conclusion Six new liver cancer differentially expressed gene fragments were obtained by DDPCR technique, indicating that the occurrence and development of liver cancer is a complex process involving multiple genes.