Detection of Differentially Expressed Genes in Glioblastoma by Suppression Subtractive Hybridization

  • Yu, Na-Mi (Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University) ;
  • Ahn, Jung-Yong (Department of Neurosurgery, Pundang CHA Hospital, College of Medicine, Pochon CHA University) ;
  • Choi, Eun-Jin (Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University) ;
  • Hong, Yong-Kil (Department of Neurosurgery, Cancer Research Institute, Catholic Research Institutes of Medical Science, The Catholic University) ;
  • Kim, Tai-Gyu (Department of Neurosurgery, Cancer Research Institute, Catholic Research Institutes of Medical Science, The Catholic University) ;
  • Kim, Chang-Hyun (Department of Neurosurgery, Cancer Research Institute, Catholic Research Institutes of Medical Science, The Catholic University) ;
  • Lee, Kyu-Sung (Department of Neurosurgery, Yonsei University College of Medicine) ;
  • Kim, Dong-Seok (Department of Neurosurgery, Yonsei University College of Medicine) ;
  • Kim, Jin-Kyeoung (Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University)
  • Published : 2005.06.28

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 Go-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 $0.5{\sim}1hr$ 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" playa role in brain tumorigenesis and the activation of this "clone 47" may be necessary for the development of cancer.

Keywords

References

  1. Chae KS, Kweon CY, Lee DK : PTEN Expression in Human Brain Glial Tumors. J Korean Neurosurg Soc 35 : 246-249, 2004
  2. Chook YM, Blobel G : Karyopherins and nuclear import. Curr Opin Struct Biol 11 : 703-715, 2001 https://doi.org/10.1016/S0959-440X(01)00264-0
  3. Christensen HC, Kosteljanetz M, Johansen C : Incidences of gliomas and meningiomas in Denmark, 1943 to 1997. Neurosurgery 52 : 1327-1333, 2003 https://doi.org/10.1227/01.NEU.0000064802.46759.53
  4. Cooper GM, Hausman RE : The Cell : A Molecular Approach, ed 3. Washington, DC : ASM press, 2004, pp631-635
  5. Dai C, Holland EC : Astrocyte differentiation states and gliomas formation. Cancer J 9 : 72-81, 2003 https://doi.org/10.1097/00130404-200303000-00002
  6. Diatchenko L, Lau YF, Campbell AP, Chenchik A, Moqadam F, Huang B, et al : Suppression subtractive hybridization : a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc Natl Acad Sci 93 : 6025-6030, 1996 https://doi.org/10.1073/pnas.93.12.6025
  7. Dong Y, Zhang H, Hawthorn L, Ganther HE, Ip C : Delineation of the molecular basis for selenium-induced growth arrest in human prostate cancer cells by oligonucleotide array. Cancer Res 63 : 52-59, 2003
  8. el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, et al : WAF1, a potential mediator of p53 tumor suppression. Cell 75 : 817-825, 1993 https://doi.org/10.1016/0092-8674(93)90500-P
  9. Han FC, Gong M, Ng HC, Ho B : Identification of H. pylori strain specific DNA sequences between two clinical isolates from NUD and gastric ulcer by SSH. World J Gastroenterol 9 : 1747-1751, 2003 https://doi.org/10.3748/wjg.v9.i8.1747
  10. Hartmann TB, Thiel D, Dummer R, Schadendorf D, Eichmuller S : SEREX identification of new tumour-associated antigens in cutaneous T-cell lymphoma. Br J Dermatol 150 : 252-258, 2004 https://doi.org/10.1111/j.1365-2133.2004.05651.x
  11. Hufton SE, Moerkerk PT, Brandwijk R, de Bruine AP, Arends JW, Hoogenboom HR : A profile of differentially expressed genes in primary colorectal cancer using suppression subtractive hybridization. FEBS Lett 463 : 77-82, 1999 https://doi.org/10.1016/S0014-5793(99)01578-1
  12. Ingman M, Kaessmann H, Paabo S, Gyllensten U : Mitochondrial genome variation and the origin of modern humans. Nature 408 : 708-713, 2000 https://doi.org/10.1038/35047064
  13. Ji W, Wright MB, Cai L, Flament A, Lindpaintner K : Efficacy of SSH PCR in isolating differentially expressed genes. BMC Genomics 3 : 12, 2002 https://doi.org/10.1186/1471-2164-3-12
  14. Kim M, Kim S, Kim S, Kim BD : Isolation of cDNA clones differentially accumulated in the placenta of pungent pepper by suppression subtractive hybridization. Mol Cells 11 : 213-219, 2001
  15. Liang P, Pardee AB : Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257 : 967-971, 1992 https://doi.org/10.1126/science.1354393
  16. Miyasaka Y, Enomoto N, Nagayama K, Izumi N, Marumo F, Watanabe M, et al : Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization. Br J Cancer 85 : 228-234, 2001 https://doi.org/10.1054/bjoc.2001.1901
  17. Nutt CL, Zerillo CA, Kelly GM, Hockfield S : Brain enriched hyaluronan binding (BEHAB)/brevican increases aggressiveness of CNS-1 gliomas in Lewis rats. Cancer Res 61 : 7056-7059, 2001
  18. Palackal NT, Lee SH, Harvey RG, Blair IA, Penning TM : Activation of polycyclic aromatic hydrocarbon trans-dihydrodiol proximate carcinogens by human aldo-keto reductase(AKR1C) enzymes and their functional overexpression in human lung carcinoma (A549) cells. J Biol Chem 277 : 24799-24808, 2002 https://doi.org/10.1074/jbc.M112424200
  19. Porkka KP, Visakorpi T : Detection of differentially expressed genes in prostate cancer by combining suppression subtractive hybridization and cDNA library array. J Pathol 193 : 73-79, 2001 https://doi.org/10.1002/1096-9896(2000)9999:9999<::AID-PATH751>3.0.CO;2-Y
  20. Rehman I, Cross SS, Azzouzi AR, Catto JW, Deloulme JC, Larre S, et al : S100A6 (Calcyclin) is a prostate basal cell marker absent in prostate cancer and its precursors. Br J Cancer 91 : 739-744, 2004 https://doi.org/10.1038/sj.bjc.6602034
  21. Sallinen SL, Sallinen PK, Haapasalo HK, Helin HJ, Helen PT, Schraml P, et al : Identification of differentially expressed genes in human gliomas by DNA microarray and tissue chip techniques. Cancer Res 60 : 6617-6622, 2000
  22. Shridhar V, Sen A, Chien J, Staub J, Avula R, Kovats S, et al : Identification of underexpressed genes in early- and late-stage primary ovarian tumors by suppression subtraction hybridization. Cancer Res 62 : 262-270, 2002
  23. Smith JM, Bowles J, Wilson M, Koopman P : HMG box transcription factor gene Hbp1 is expressed in germ cells of the developing mouse testis. Dev Dyn 230 : 366-370, 2004 https://doi.org/10.1002/dvdy.20053
  24. Swearingen ML, Sun D, Bourner M, Weinstein EJ : Detection of differentially expressed HES-6 gene in metastatic colon carcinoma by combination of suppression subtractive hybridization and cDNA library array. Cancer Lett 198 : 229-239, 2003 https://doi.org/10.1016/S0304-3835(03)00313-6
  25. Tanaka T, Yamakawa N, Usui M : Level of glucocorticoid receptor mRNA expression in iris tissue and postoperative ocular inflammation in patients with uveitis. Graefes Arch Clin Exp Ophthalmol 241 : 81-84, 2003 https://doi.org/10.1007/s00417-002-0593-5
  26. Van Nocker S, Ludwig P : The WD-repeat protein superfamily in Arabidopsis : conservation and divergence in structure and function. BMC Genomics 4 : 50, 2003 https://doi.org/10.1186/1471-2164-4-50
  27. Vasmatzis G, Essand M, Brinkmann U, Lee B, Pastan I : Discovery of three genes specifically expressed in human prostate by expressed sequence tag database analysis. Proc Natl Acad Sci 95 : 300-304, 1998 https://doi.org/10.1073/pnas.95.1.300
  28. Velculescu VE, Zhang L, Zhou W, Vogelstein J, Basrai MA, Bassett DE Jr, et al : Characterization of the yeast transcriptome. Cell 88 : 243-251, 1997 https://doi.org/10.1016/S0092-8674(00)81845-0
  29. Wandrey M, Trevaskis B, Brewin N, Udvardi MK : Molecular and cell biology of a family of voltage-dependent anion channel porins in Lotus japonicus. Plant Physiol 134 : 182-193, 2004 https://doi.org/10.1104/pp.103.031484
  30. Welsh J, Chada K, Dalal SS, Cheng R, Ralph D, McClelland M : Arbitrarily primed PCR fingerprinting of RNA. Nucleic Acids Res 20 : 4965-4970, 1992 https://doi.org/10.1093/nar/20.19.4965
  31. Zeindl-Eberhart E, Haraida S, Liebmann S, Jungblut PR, Lamer S, Mayer D, et al : Detection and identification of tumor-associated protein variants in human hepatocellular carcinomas. Hepatology 39 : 540-549, 2004 https://doi.org/10.1002/hep.20060