癌变·畸变·突变 ›› 1999, Vol. 11 ›› Issue (1): 8-010.doi: 10.3969/j.issn.1004-616x.1999.01.003

• 论著 • 上一篇    下一篇

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

杨梅英 叶常青 陈剑云 刘雷华 袁 雄   

  1. 北京放射医学研究所 北京 100850
  • 收稿日期:1998-08-19 修回日期:1998-09-30 出版日期:1999-01-30 发布日期:1999-01-30

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

Yang Meiying, Ye Changqing, Liu Leihua   

  1. Institute of Radiation Medicine, Beijing  100850
  • Received:1998-08-19 Revised:1998-09-30 Online:1999-01-30 Published:1999-01-30

摘要: 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,为确定相关基因奠定了基础。

关键词: 差异显示, 放射自显影, 硝影银染色, 辐射致癌

Abstract: 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 f rom two pairs of sample , SV40 - immortalized human fetal t racheal fibroblast cell (SHTF) versusαSHTF cell (SHTF cell irradiated byαparticle) , and lung cancer tissue versus normal lung tissue obtained f rom one patient . These RNAs were amplified by RTPCR. The differential expressed gene fragment s were display by autoradiograph or silver nit rate stain. The result showed that the differential display mRNA were established on both cell and tissue samples. The bands stained by silver nit rate is more clear than the bands on X2ray film. The rate of reamplification of differential expressed genes f ragment s stained by silver nit rate is 80 % , higher than autoradiograph , 50 %.

Key words: Differential Display mRNA, Autoradiograph, Silver nit rate stain, Cancer induced by irradiation