Objective: To clone apoptosis-related genes induced by all-trans retinoic acid (ATRA) from human breast cancer cell line MCF-7, and to analyze the association between the cloned genes and apoptosis.
Methods: An apoptotic cell model of MCF-7 cells was constructed with ATRA induction. The apoptosis-related genes were cloned by improved PCR-based subtractive hybridization.
Results: Twelve differentially expressed clones were screened out. After exclusion of false positive clones by reverse dot blotting, 5 clones containing fragments of 0.5-1.5 kb were sequenced. The results of sequencing were compared with BLAST. A novel gene, named apmcf-1, coding for 47 amino acids was identified. This gene was accepted by GenBank, (Accession number: AF141882). The result of reverse dot blotting showed that it was related to apoptosis. The other 4 genes were already known. Three of them were related to apoptosis as previously reported. Two of them, hsp-90 and rb-L3, had little information about their relationship with apoptosis.
Conclusion: ATRA-induced tumor cell apoptosis is a complex process with multiple genes involved.