Feng L, Wang X Q, Fu M, Wang Z H, Tian Y, Cai Y, Wu M
Department of Cell Biology, Cancer Institute, Chinese Academy of Medical Sciences, Beijing.
Sci China B. 1992 Apr;35(4):445-54.
Treatment of the human esophageal cancer cell line EC8712 with retinoic acid (RA) stopped the cell growth significantly and gave rise to terminal differentiation of the cells characterized by increased expression of involucrin gene. Two cDNA libraries were constructed from the parental and RA-treated cells respectively. Repeated subtractive hybridization of single-stranded plasmid DNA prepared from pooled colonies of cDNA library of the parental cells with cDNA probe generated from the RA-treated cells exhausted sequences common to both libraries of the cell. The unhybridized cDNA probe represented, therefore, the genes activated after RA-treatment. By using these enriched cDNAs as probe to screen the cDNA library constructed from the RA-treated cells thirty-nine positive colonies were obtained, of which two were specifically due to RA-induction. One of these two cDNA clones, designated as pRA538, has undergone further analysis and shown differentiation-inducing effect on parental cancer cells. A novel strategy for cloning genes involved in terminal differentiation of cancer cells is developed.
用视黄酸（RA）处理人食管癌细胞系EC8712可显著抑制细胞生长，并导致细胞终末分化，其特征是外皮蛋白基因表达增加。分别从亲代细胞和经RA处理的细胞构建了两个cDNA文库。用经RA处理的细胞产生的cDNA探针与从亲代细胞cDNA文库的汇集菌落制备的单链质粒DNA进行反复消减杂交，耗尽了两个细胞文库共有的序列。因此，未杂交的cDNA探针代表了RA处理后激活的基因。用这些富集的cDNA作为探针筛选从经RA处理的细胞构建的cDNA文库，获得了39个阳性菌落，其中两个是RA诱导特异性产生的。这两个cDNA克隆之一命名为pRA538，已进行进一步分析，并显示对亲代癌细胞有诱导分化作用。开发了一种克隆参与癌细胞终末分化的基因的新策略。
