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Journal of Korean Neurosurgical Society > Volume 37(6); 2005 > Article
Journal of Korean Neurosurgical Society 2005;37(6): 443-448.
Detection of Differentially Expressed Genes in Glioblastoma by Suppression Subtractive Hybridization.
Na Mi Yu, Jung Yong Ahn, Eun Jin Choi, Yong Kil Hong, Tai Gyu Kim, Chang Hyun Kim, Kyu Sung Lee, Dong Seok Kim, Jin Kyeoung Kim
1Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University, Seongnam, Korea. kyeoung@cha.ac.kr
2Department of Neurosurgery, Pundang CHA Hospital, College of Medicine, Pochon CHA University, Seongnam, Korea.
3Department of Neurosurgery, Cancer Research Institute, Catholic Research Institutes of Medical Science, The Catholic University, Seoul, Korea.
4Department of Neurosurgery, Yonsei University College of Medicine, Seoul, Korea.
ABSTRACT
OBJECTIVE:
A variety of genetic alterations in human glioblastoma comprises signal transduction and cell cycle arrest control of cellular processes. Subtractive hybridization is potentially a faster method for identifying differentially expressed genes associated with a particular disease state. Using the technique of subtraction, we isolated novel genes that are overexpressed in glioblastoma tissue as compared to normal brain tissue.
METHODS:
We evaluated the differential expression of genes in each of hybridizing tester and driver cDNAs to digested 130 clones. After sequencing of 130 clones and homology search, this study performed to determine mRNA expression of the unknown gene, "clone 47", in brain tissue, glioblasoma, and several cancer cell lines by reverse transcription-polymerase chain reaction (RT-PCR). To test the time course for G0-phase arrest, serum stimulation and expression at various times for RT-PCR performed.
RESULTS:
We identified 23 novel genes by BLAST of the digested 130 clones. The expressions of "clone 47" mRNA of glioblastoma and several cancer lines were significantly higher than normal brain tissues and several normal cell lines. We confirmed the mRNA expression of "clone 47" was up-regulation for 0.5~1hr of WI-38 cell differentiation.
CONCLUSION:
The novel gene, "Clone 47" is upregulated in glioblastoma tissue and several cancer cell lines. This gene is time dependent activation during time course of serum stimulation. This result suggests that "clone 47" play a role in brain tumorigenesis and the activation of this "clone 47" may be necessary for the development of cancer.
Key Words: Genes; Glioblastoma; Tumorigenesis; Suppression subtractive hybridization
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