[Apoptosis-related genes cloned by improved subtractive hybridization].
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.


CONCLUSIONS
ATRA-induced tumor cell apoptosis is a complex process with multiple genes involved.
Yan
W
W
Department of Pathology, Fourth Military Medical University, Xi'an, 710032, China.
Li
Q
Q
Zhu
F
F
D017209:Apoptosis / Q000235:genetics*; D001943:Breast Neoplasms; D003001:Cloning, Molecular / Q000379:methods*; D019204:GTP-Binding Proteins; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D011518:Proto-Oncogene Proteins; C449560:SRPRB protein, human; D014212:Tretinoin; D019204:GTP-Binding Proteins