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J Korean Soc Ther Radiol Oncol > Volume 17(2); 1999 > Article
The Journal of the Korean Society for Therapeutic Radiology and Oncology 1999;17(2): 151-157.
Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line
Jae Chul Kim, In Kyu Park, Kyu Bo Lee, Sang Kyun Sohn, Moo Kyu Kim, Jung Chul Kim
1Department of Radiation Oncology, School of Medicine, Kyungpook National University, Taegu, Korea.
2Department of Internal Medicine, School of Medicine, Kyungpook National University, Taegu, Korea.
3Department of Immunology, School of Medicine, Kyungpook National University, Taegu, Korea.
ABSTRACT
PURPOSE:
By sequencing the Expressed Sequence Tags of human dermal papilla cDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL60 cell line.
MATERIALS AND METHODS:
K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusion expression system and the protein product was identified on SDS-PAGE.
RESULTS:
K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares 70% identity with that of rat glutathione S-transferase kappa 1 (rGSTK1). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in colorectal cancer and melanoma cell lines.
CONCLUSION:
Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.
Key Words: Glutathione S-Transferase, Radioresistance, HL60 cell
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