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Characterization of colorectal-cancer-related cDNA clones obtained by subtractive hybridization screening

  • Original Paper
  • Experimental Oncology
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Abstract

In an attempt to seek out new factors that are related to colorectal carcinogenesis at the molecular level, subtractive hybridization between cDNA of normal mucosal tissues and mRNA of colorectal carcinoma tissues was performed. Subsequent screenings of the cDNA libraries, constructed from normal mucosal tissues, using the “subtractive probes” generated a total of 46 clones that were expressed in normal mucosa but were either expressed at a significantly reduced level or not expressed at all in cancer tissues. Partial nucleotide sequences of all of these cDNA clones were determined, and sequence homology analyses were performed with the Genbank database. Of the 46 cDNA samples, 44 contained substantial sequence homologies with 32 immunoglobulin gene fragments, a helix-loop-helix basic phosphoprotein gene, an acidic ribosomal phosphoprotein P2 gene, aBLR1 gene for Burkitt's lymphoma receptor 1 gene, D5S419 DNA segment containing (C-A) repeats, a glucokinase (GCK) gene, a Na+, K+-ATPase α-subunit gene, a histocompatibility system HLA-DR heavy-chain gene, a dystrophic gene, a mucin (MUC2) gene, a μ-glutathioneS-transferase gene, a Menkes disease protein gene, and a 40-kDa keratin intermediate filament precursor gene. The remaining two cDNA clones (now registered under GenBank accession numbers U17714 and U20428) showed few (less than 60%) sequence homologies with any known sequences in the GenBank database and, therefore, may represent novel genes whose expression was down-regulated in human colorectal carcinomas. The possible clinical significance of these findings and the involvement of these two genes in the carcinogenesis of colorectal as well as other cancers are being investigated.

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References

  • Benton WD, Davis RW (1997) Screening lambda gt recombinant clones by hybridization to single plaques in situ. Science 196:180–182

    Google Scholar 

  • Eckert RL (1988) Sequence of the human 40-kDa keratin reveals an unusual structure with very high sequence identity to the corresponding bovine keratin. Proc Natl Acad Sci USA 85:1114–1118

    PubMed  Google Scholar 

  • Elvin P, Kerr IB, Mcardle CS, Birnie GD (1988) Isolation and preliminary characterisation of cDNA clones representing mRNAs associated with tumor progression and metastasis in colorectal cancer. Br J Cancer 57:36–42

    PubMed  Google Scholar 

  • Fan J, Devesa SS, Zheng W, Blot WJ, Fraumeni JF Jr, Gao YT (1993) Cancer incidence trends in urban Shanghai, 1972–1989. Int J Cancer 53:764–770

    PubMed  Google Scholar 

  • Fargnoli J, Holbrook NJ, Fornace AJ Jr (1990) Low-ratio hybridization subtraction. Anal Biochem 187:364–373

    PubMed  Google Scholar 

  • Fearon ER, Vogelstein B (1990) A genetic model for colorectal tumorigenesis. Cell 61:759–767

    PubMed  Google Scholar 

  • Hamilton SR (1992) Molecular genetics of colorectal carcinoma. Cancer 70:1216–1221

    PubMed  Google Scholar 

  • Modrich P (1994) Mismatch repair, genetic stability and cancer. Science 266:1959–1960

    PubMed  Google Scholar 

  • Moll R, Franke WW, Schiller DL, Geiger B, Krepler R (1982) The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 31:11–24

    PubMed  Google Scholar 

  • Pao C, Lin CY, Yao DS, Tseng CJ (1995) Differential expression of cytokine gene in cervical cancer tissues. Biochem Biophy Res Commun 214:1146–1151

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Takemura K, Hirokawa K, Esaki Y, Mishima Y (1990) Distribution of immunoglobulins and secretory component in gastric cancer of the aged. Cancer 66:2168–2173

    PubMed  Google Scholar 

  • Tew KD (1994) Glutathione-associated enzymes in anticancer drug resistance. Cancer Res 54:4313–4320

    PubMed  Google Scholar 

  • Zheng S (1993) Application of subtractive hybridization in screening for colorectal cancer related genes. Chin Med Sci J 6:100

    Google Scholar 

  • Zheng S, Cai X, Cao J, Zheng L, Geng L, Zhang Y, Gu J, Shi Z (1997) Screening and identification of down-regulated genes in colorectal carcinoma by subtractive hybridization: a method to identify putative tumor suppresser genes. Chin Med J 110 (in press)

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This work was supported by research grants from the chinese National 8th 5-Year medical Strategic Science and technology Plan (85-914-01-09), the National Science foundation of China (39170800 and 39340007), and the Zhejiang Provincial Science Foundation (393041)

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Cao, J., Cai, X., Zheng, L. et al. Characterization of colorectal-cancer-related cDNA clones obtained by subtractive hybridization screening. J Cancer Res Clin Oncol 123, 447–451 (1997). https://doi.org/10.1007/BF01372549

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  • DOI: https://doi.org/10.1007/BF01372549

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