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[Apoptosis-related genes cloned by improved subtractive hybridization]
1Department of Pathology, Fourth Military Medical University, Xi'an, 710032, China.
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.
Insights
All-trans retinoic acid (ATRA) induces apoptosis in human breast cancer cells. Researchers identified a novel apoptosis-related gene, apmcf-1, and four known genes involved in this complex process.
Area of Science:
- Molecular biology
- Cancer research
- Gene expression analysis
Context:
- Investigating the molecular mechanisms of all-trans retinoic acid (ATRA) in inducing apoptosis in human breast cancer cell line MCF-7.
- Understanding the genetic basis of ATRA-mediated apoptosis is crucial for developing targeted cancer therapies.
Purpose:
- To identify and clone novel apoptosis-related genes induced by ATRA in MCF-7 cells.
- To analyze the functional association of these cloned genes with the process of apoptosis.
Summary:
- Differential gene expression was analyzed using improved PCR-based subtractive hybridization in ATRA-treated MCF-7 cells.
- A novel gene, apmcf-1 (GenBank Accession: AF141882), was identified and confirmed to be related to apoptosis.
- Four known genes, including three previously linked to apoptosis and two with limited information (hsp-90, rb-L3), were also identified.
Impact:
- This study reveals the complexity of ATRA-induced apoptosis, highlighting the involvement of multiple genes.
- The identification of apmcf-1 provides a new target for further research into breast cancer treatment strategies.