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Original Article
- Screening and Cloning of Genes Preferentially Expressed in Aminopterin-Treated Myeloma Cell Apoptosis
- Hong Chan Lee, Sung Joo Kim, Sarah Yoon, Sung Joon Kwon, Yong Hoon Chung
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Journal of the Korean Cancer Association 2000;32(3): 655-664.
1Departments of General Surgery, College of Medicine, Hanyang University, Seoul, Korea.
2Departments of Microbiology, College of Medicine, Hanyang University, Seoul, Korea.
2Departments of Microbiology, College of Medicine, Hanyang University, Seoul, Korea.
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Abstract
PURPOSE
When murine myeloma cells P3-X63-Ag8.653 (V653) of this model treated with amin opterin, an anticancer drug, they can't synthesize nucleic acid via de novo or salvage pathway and selectively eliminated due to apoptosis. This study was aimed to clone specific known and novel genes preferentially expressed in aminopteirn-treated tumor cell apoptosis.
MATERIALS AND METHODS
This study was aimed to clone specific known and novel genes pre ferentially expressed in aminopteirn-treated tumor cell apoptosis by using subtraction-PCR technique.
RESULTS
By using this technique 868 clones were obtained.
Of these 427 clones were positive with insert DNA. By using cross-hybridization Southern blotting, final 101 clones were selected.
All of these genes were sequenced and analyzed by using genebank DNA database. Total 101 clones of genes preferentially expressed in apoptotic tumor cells were classified into 10 groups, which included ribosomal proteins, nuclear proteins, mitdegrees Chondrial proteins, signal transductional proteins, retroviral proteins, cell surface receptor proteins, cell structural proteins, unclassified miscellaneous proteins, and novel genes. Especially, Unknown novel genes preferentially ex pressed in this apoptotic tumor cells included clone numbers S1-63, 1-1, 1-3, 1-16, 1-18, 1-20, 3-33, 3-41, 3-44, 3-48, 3-55, 3-60, 6-17, 6-25, 8-12, 50-7, 50-23, and 100-35.
CONCLUSION
It seemed that known and unknown novel genes cloned in this study would con tribute to the future studies regarding apoptosis of tumor cells and cancer treatment therepy.
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