差异显示法的建立及其在辐射致癌机理研究中的初步应用

ESTABLISHMENT OF DIFFERENTIAL DISPLAY mRNA AND THE APPLICATION IN THE STUDY ON MECHANISM OF LUNG CANCER INDUCED BY IRRADIATION

本研究目的建立差异显示技术，将其应用于辐诱发肺癌发生相关基因研究中。取ＳＨＴＦ细胞（ＳＶ４０永生化人胎气管成纤维细胞）和αＳＨＴＦ细胞（α粒子照射后能在软琼脂上形成克隆的ＳＨＴＦ细胞）以及肺癌组织和同例正常肺组织为模板，将其ｍＲＮＡ进行ＲＴＰＣＲ扩增，采用放射自显影法和硝酸银染色法显示扩增片段，并比较两者的显示效果。结果表明：在细胞和组织样品中均建立了差异显示法，但硝酸银染色的条带较放射自显影显示的清晰，再扩增后的显示率也较高，两者的复显示分别为８０％和５０％。结论：在细胞和组织样品中成功地建立了差异显示法，改进的条带显示法提高了复显率。

This report describes a method to separate mRNAs by means of the Polymerase Chain Reaction(Differential Display mRNA), and identifies the genes related to lung cancer induced by radiation by this method. The RNAs were isolated from two pairs of sample, SV40-immortalized human fetal tracheal fibroblast cell (SHTF) versus αSHTF cell (SHTF cell irradiated by α particle), and lung cancer tissue versus normal lung tissue obtained from one patient. These RNAs were amplified by RTPCR. The differential expressed gene fragments were display by autoradiograph or silver nitrate stain. The result showed that the differential display mRNA were established on both cell and tissue samples. The bands stained by silver nitrate is more clear than the bands on X ray film. The rate of reamplification of differential expressed genes fragments stained by silver nitrate is 80%, higher than autoradiograph, 50%.