Liang P, Averboukh L, Keyomarsi K, Sager R, Pardee A B
Division of Cell Growth and Regulation, Dana-Farber Cancer Institute, Boston, Massachusetts 02115.
Cancer Res. 1992 Dec 15;52(24):6966-8.
Identification of the genes that are specifically expressed in tumor cells but not in normal cells (oncogenes), or vice versa (tumor suppressor genes), is important for understanding the molecular basis of cancer. The differential display technique was applied to compare mRNAs from normal and tumor-derived human mammary epithelial cells, cultured under the same conditions. Complementary DNA fragments corresponding to several apparently differentially expressed mRNAs were recovered and sequenced. They exhibit characteristics of the 3' end of eukaryotic mRNA, as predicted by the method. A complementary DNA fragment seen only in the normal cell was used as a probe to isolate its corresponding complementary DNA clone from a library. Northern analysis confirmed its differential expression. Thus, this method can be used for detecting, cloning, and sequencing of genes that are unique to a host of biological and disease processes.
鉴定在肿瘤细胞中特异性表达而在正常细胞中不表达的基因(癌基因),或者反之(肿瘤抑制基因),对于理解癌症的分子基础很重要。应用差异显示技术来比较在相同条件下培养的正常和肿瘤来源的人乳腺上皮细胞的mRNA。回收并测序了对应于几种明显差异表达的mRNA的互补DNA片段。正如该方法所预测的,它们表现出真核mRNA 3'端的特征。仅在正常细胞中出现的一个互补DNA片段被用作探针,从文库中分离其相应的互补DNA克隆。Northern分析证实了其差异表达。因此,该方法可用于检测、克隆和测序一系列生物和疾病过程所特有的基因。