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In situ hybridization as a tool to study numerical chromosome aberrations in solid bladder tumors

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Summary

Methods for single- and double-target in situ hybridization (ISH) to, cells isolated from solid transitional cell carcinomas (TCC's) of the urinary bladder are described. Single cell suspensions were prepared from solid tumors of the urinary bladder by mechanical disaggregation and fixed in 70% ethanol. Using two DNA probes specific for the centromeres of chromosomes #1 and #18, ISH procedures were optimized for these samples. Human lymphocytes and cells from the T24 bladder tumor cell line were used as controls. In lymphocyte nuclei and metaphase chromosome spreads, ISH showed two major spots for each of the probes. About 80% of the nuclei from T24 cells showed three spots for both the chromosome #1 and #18 specific probes. When nuclei from TCC's were analyzed, often the number of spots for chromosome #1, and to a lesser extent for chromosome #18, differed from the number expected on basis of flow cytometric ploidy measurements. The double target-ISH method in all cases allowed the correlation of numerical aberrations for chromosomes #1 and #18 in one and the same cell. By such analyses a profound heterogeneity in chromosome number was detected in most tumors. In order to optimize the reproductbility of the method and the interpretation of the ISH-signals, criteria for their analysis have been determined. This procedure can now be applied on a routine basis to solid tumor specimens.

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References

  • Atkin NB (1986) Chromosome 1 aberrations in cancer. Cancer Genet Cytogenet 21:279–285

    Google Scholar 

  • Atkin NB, Baker MC (1985) Cytogenetic study of ten carcinomas of the bladder: Involvement of chromosomes 1 and 11. Cancer Genet Cytogenet 15:253–268

    Google Scholar 

  • Bock G, Hilchenbach M, Schauenstein K, Wick G (1985) Photometric analysis of antifading reagents for immunofluorescence with laser and convential illumination sources. J Histochem Cytochem 33:699–705

    Google Scholar 

  • Brigati DJ, Myerson D, Leary JJ, Spalholz B, Travis S, Fong CKY, Hsiung GD, Ward DC (1982) Detection of viral genomes in cultured cells and paraffin-embedded tissues sections using biotin-labeled hybridization probes. Virology 126:32–50

    Google Scholar 

  • Bubeník J, Baresôvá V, Viklický V, Jakoubková J, Sainerová H, Donner J (1973) Established cell line of urinary bladder carcinoma (T24) containing tumour-specific antigen. Int J Cancer 11:765–733

    Google Scholar 

  • Cooke HJ, Hindley J (1979) Cloning of human satellite III DNA: different components are on different chromosomes. Nucleic Acid Res 6:3177–3197

    Google Scholar 

  • Cremer T, Landegent J, Brückner A, Scholl HP, Schardin M, Hager HD, Devilee P, Pearson P, van der Ploeg M (1986) Detection of chromosome aberrations in the human interphase nucleus by visualization of specific target DNAs with radioactive and non-radioactive in situ hybridization techniques: diagnosis of trisomy 18 with probe L1.84. Hum Genet 74:346–352

    Google Scholar 

  • Cremer T, Tessin D, Hopman AHN, Manuelidis L (1988) Rapid interphase and metaphase assessment of specific chromosomal changes in neuroectodermal tumor cells by in situ hybridization with chemically modified DNA probes. Exp Cell Res (in press)

  • Cummings KB (1980) Carcinoma of the bladder: Predictors. Cancer 45:1849–1855

    Google Scholar 

  • Cutler SJ, Heney NM, Friedell GH (1982) Longitudial study of patients with bladder cancer: factors associated with disease recurrence and progression. In: Bonney WW, Prout GR (ed) AUA monographs, bladder cancer, vol 1, Chap 4. Williams and Wilkins, Baltimore, pp 35–46

    Google Scholar 

  • Devilee P, Cremer T, Slagboom P, Bakker E, Scholl HP, Hager HD, Stevenson AFG, Cornelisse CJ, Pearson PL (1986) Two subsets of human alphoid repetitive DNA show distinct preferential localization in the pericentromeric heterochromatin of chromosome 13, 21, 18. Cytogenet Cell Genet 41:193–201

    Google Scholar 

  • Devonec M, Hijazi A (1985) A new concept in the natural history of human bladder cancer based on genetic instability of tumors as evidenced by flow cytometric DNA analysis. 2nd. French Congress of cytometry, Rennes

  • Feitz WFJ, Beck HLM, Smeets AWGB, Debruyne FMJ, Vooijs GP, Herman CJ, Ramaekers FCS (1985) Tissue specific markers in flow cytometry of urological cancers: cytokeratins in bladder carcinoma. Int J Cancer 36:349–356

    Google Scholar 

  • Granberg-Öhman I, Tribukait B, Wijkström H, Berlin T, Collste L (1980) Papillary carcinoma of the urinary bladder. A study of chromosomal and cytofluorometric DNA analysis. Urol Res 8:87–93

    Google Scholar 

  • Hopman AHN, Wiegant J, Tesser GI, Duijn P van (1986a) A non-radioactive in situ hybridization method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands. Nucleic Acids Res 14:6471–6488

    Google Scholar 

  • Hopman AHN, Wiegant J, Duijn P van (1986b) A new hybridocytochemical method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands. I. Stability of the mercury-sulfhydryl bond and influence of the ligand structure of the immunochemical detection of the hapten. Histochemistry 84:169–178

    Google Scholar 

  • Hopman AHN, Wiegant J, Raap AK, Landegent JE, Van der Ploeg M, Van Duijn P (1986c) Bi-color detection of two target DNAs by non-radioactive in situ hybridization. Histochemistry 85:1–4

    Google Scholar 

  • Hopman AHN, Wiegant J, Duijn P van (1987) Mercurated nucleic acid probes, a new principle for non-radioactive in situ hybridization. Exp Cell Res: 169:357–368

    Google Scholar 

  • Kovacs G (1985) Serial cytogenetic analysis in a patient with pseudodiploid bladder cancer. J Cancer Res Clin Oncol 110:249–251

    Google Scholar 

  • Landegent JE, Jansen in de Wal N, Baan RA, Hoeijmakers JHJ, Ploeg M van der (1984) 2-acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences. Exp Cell Res 153:61–72

    Google Scholar 

  • Landegent JE, Jansen in de Wal N, Ploem JS, Ploeg M van der (1985) Sensitive detection of hybridocytochemical results by means of reflection-contrast microscopy. J Histochem Cytochem 33:1241–1246

    Google Scholar 

  • Langer PR, Waldrop AA, Ward DA (1981) Enzymatic synthesis of biotin labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci USA 78:6633–6637

    Google Scholar 

  • Levi PE, Cooper EH, Anderson CK, Path MC, Williams RE (1969) Analysis of DNA content, nuclear size and cell proliferation of transitional cell carcinoma in man. Cancer 23:1074–1085

    Google Scholar 

  • Nederlof PM, Raap AK, Robinson D, Wiegant J, Hopman AHN, Tanke HJ (1987) Simultaneous detection of three and more target DNA sequences by three color fluorescence in situ hybridization. Eur J Cell Biol (Suppl 20) 44:97

    Google Scholar 

  • Raap AK, Marijnen JGJ, Vrolijk J, Ploeg M van der (1986) Denaturation, renaturation, and loss of DNA during in situ hybridization procedures. Cytometry 7:235–242

    Google Scholar 

  • Sandberg AA (1977) Chromosome markers and progression in bladder cancer. Cancer Res 37:222–229

    Google Scholar 

  • Singer M (1982) Highly repeated sequences in mammalian genomes. Int Rev Cytol 76:67–112

    Google Scholar 

  • Smeets AWGB, Pauwels RPE, Beck HLM, Geraedts JPM, Debruyne FMJ, Laarakkers L, Feitz WFJ, Vooijs GP, Ramaekers FCS (1987a) Tissue specifc markers in flow cytometry of urological cancers III. Comparing chromosomal and flow cytometric analysis of bladder tumors. Int J Cancer 39:304–310

    Google Scholar 

  • Smeets W, Pauwels R, Laarakkers L, Debruyne F, Geraedts J (1987b) Chromosomal analysis of bladder cancer III. Nonrandom alterations. Cancer Genet Cytogen 29:29–41.

    Google Scholar 

  • Smeets AWGB, Pauwels RPE, Beck HLM, Feitz WFJ, Geraedts JPM, Debruyne FMJ, Laarakkers L, Vooijs GP, Ramaekers FCS (1987c) Comparison of tissue disaggregation techniques of transitional cell bladder carcinomas for flow cytometry and chromosomal analysis. Cytometry 8:14–19

    Google Scholar 

  • Summers JL, Falor WH, Ward R (1981) A 10-year analysis of chromosomes in non-invasive papillary carcinoma of the bladder. J Urol 125:177–178

    Google Scholar 

  • Tavares AS, Costa J, Carvalho A, de Reis M (1980) Tumour ploidy and prognosis in carcinomas of the bladder and prostate. In: Sandberg AA (ed) The chromosomes in human cancer and leukemia. Elsevier-North Holland, New York, pp 458–516

    Google Scholar 

  • Tribukait B, Granberg-Öhman I, Wijkström H (1986) Flow cytometric DNA and cytogenetic studies in human tumors: A comparison and discussion of the differences in modal values obtained by two methods. Cytometry 7:194–199

    Google Scholar 

  • Vanni R, Peretti D, Scarpa RM, Usai E (1985) Derivative 11 marker chromosome in bladder carcinoma. Cancer Genet Cytogenet 16:289–295

    Google Scholar 

  • Wijkström H, Granberg-Öhman I, Tribukait B (1984) Chromosomal and DNA patterns in transitional cell bladder carcinoma. A comparative cytogenetic and flow-cytofluorometric DNA study. Cancer 53:1718–1723

    Google Scholar 

  • Willard FW, Waye JS (1987) Hierarchical order in chromosomespecific human alpha satellite DNA. TIG 3:192–198

    Google Scholar 

  • Willard HF, Smith KD, Sutherland J (1983) Isolation and characterization of a tandem repeat family from the human X chromosome. Nucleic Acids Res 11:2017–2033

    Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, University Hospital Nijmegen, Geert Grooteplein Zuid 24, 6525 GA, Nijmegen, The Netherlands

    A. H. N. Hopman, F. C. S. Ramaekers, J. L. M. Beck & G. P. Vooijs

  2. Department of Cytochemistry and Cytometry, University of Leiden, Wassenaarseweg 72, 2333 AL, Leiden, The Netherlands

    A. K. Raap & M. van der Ploeg

  3. Department of Anthropogenetics, University of Leiden, Wassenaarseweg 72, 2333 AL, Leiden, The Netherlands

    P. Devilee

Authors
  1. A. H. N. Hopman
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  2. F. C. S. Ramaekers
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  3. A. K. Raap
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  4. J. L. M. Beck
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  5. P. Devilee
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  6. M. van der Ploeg
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  7. G. P. Vooijs
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Hopman, A.H.N., Ramaekers, F.C.S., Raap, A.K. et al. In situ hybridization as a tool to study numerical chromosome aberrations in solid bladder tumors. Histochemistry 89, 307–316 (1988). https://doi.org/10.1007/BF00500631

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