Chronic lymphocytic leukemia cells with allelic deletions at 13q14 commonly have one intact RB1 gene: evidence for a role of an adjacent locus (original) (raw)

Abstract

We have previously shown that 30% of patients with B-cell chronic lymphocytic leukemia (B-CLL) have hemizygous deletions of the retinoblastoma (RB1) gene at 13q14. RB1 gene deletions may thus participate in malignant transformation of B-CLL, but it is also possible that a neighboring gene on 13q is the relevant one. To answer this question the remaining RB1 allele of eight clones with hemizygous deletions was studied by reverse transcription-polymerase chain reaction (RT-PCR), single-strand conformation polymorphism (SSCP) analysis, and immunofluorescense techniques. Cells from 10 patients without RB1 gene deletions were also studied by these methods. Lack of RB1 mRNA and RB protein expression was seen in leukemia cells from one of the patients. All other cases were found to be normal with regard to immunofluorescense, RT-PCR, and SSCP analysis, indicating at least one functional RB1 allele and supporting the importance of another gene in the 13q14 deletions. We then performed extended Southern blot analyses of the 13q region, using probes for 10 different loci. In 14 of 31 CLL clones (45%), deletions of a region telomeric to the RB1 gene (D13S25) were observed. In 4 of the cases the deletions were homozygous. Hemizygous deletions of the RB1 gene were observed in 11 of these patients and in none of the patients without D13S25 deletions. These data thus indicate that a gene(s) telomeric to RB1 is involved in the malignant transformation of CLL clones and that deletions of this region are a common event in this disease.

8697

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andersson B., Fält S., Lambert B. Strand specificity for mutations induced by (+)-anti BPDE in the hprt gene in human T-lymphocytes. Mutat Res. 1992 Sep;269(1):129–140. doi: 10.1016/0027-5107(92)90168-2. [DOI] [PubMed] [Google Scholar]
  2. Benedict W. F., Xu H. J., Hu S. X., Takahashi R. Role of the retinoblastoma gene in the initiation and progression of human cancer. J Clin Invest. 1990 Apr;85(4):988–993. doi: 10.1172/JCI114575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown A. G., Ross F. M., Dunne E. M., Steel C. M., Weir-Thompson E. M. Evidence for a new tumour suppressor locus (DBM) in human B-cell neoplasia telomeric to the retinoblastoma gene. Nat Genet. 1993 Jan;3(1):67–72. doi: 10.1038/ng0193-67. [DOI] [PubMed] [Google Scholar]
  4. Caspersson T., Lomakka G., Zech L. The 24 fluorescence patterns of the human metaphase chromosomes - distinguishing characters and variability. Hereditas. 1972;67(1):89–102. doi: 10.1111/j.1601-5223.1971.tb02363.x. [DOI] [PubMed] [Google Scholar]
  5. Dunn J. M., Phillips R. A., Zhu X., Becker A., Gallie B. L. Mutations in the RB1 gene and their effects on transcription. Mol Cell Biol. 1989 Nov;9(11):4596–4604. doi: 10.1128/mcb.9.11.4596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Einhorn S., Burvall K., Juliusson G., Gahrton G., Meeker T. Molecular analyses of chromosome 12 in chronic lymphocytic leukemia. Leukemia. 1989 Dec;3(12):871–874. [PubMed] [Google Scholar]
  7. Fitchett M., Griffiths M. J., Oscier D. G., Johnson S., Seabright M. Chromosome abnormalities involving band 13q14 in hematologic malignancies. Cancer Genet Cytogenet. 1987 Jan;24(1):143–150. doi: 10.1016/0165-4608(87)90091-4. [DOI] [PubMed] [Google Scholar]
  8. Friend S. H., Bernards R., Rogelj S., Weinberg R. A., Rapaport J. M., Albert D. M., Dryja T. P. A human DNA segment with properties of the gene that predisposes to retinoblastoma and osteosarcoma. Nature. 1986 Oct 16;323(6089):643–646. doi: 10.1038/323643a0. [DOI] [PubMed] [Google Scholar]
  9. Hayashi K. PCR-SSCP: a simple and sensitive method for detection of mutations in the genomic DNA. PCR Methods Appl. 1991 Aug;1(1):34–38. doi: 10.1101/gr.1.1.34. [DOI] [PubMed] [Google Scholar]
  10. Horowitz J. M., Yandell D. W., Park S. H., Canning S., Whyte P., Buchkovich K., Harlow E., Weinberg R. A., Dryja T. P. Point mutational inactivation of the retinoblastoma antioncogene. Science. 1989 Feb 17;243(4893):937–940. doi: 10.1126/science.2521957. [DOI] [PubMed] [Google Scholar]
  11. Juliusson G., Oscier D. G., Fitchett M., Ross F. M., Stockdill G., Mackie M. J., Parker A. C., Castoldi G. L., Guneo A., Knuutila S. Prognostic subgroups in B-cell chronic lymphocytic leukemia defined by specific chromosomal abnormalities. N Engl J Med. 1990 Sep 13;323(11):720–724. doi: 10.1056/NEJM199009133231105. [DOI] [PubMed] [Google Scholar]
  12. Kaye F. J., Kratzke R. A., Gerster J. L., Horowitz J. M. A single amino acid substitution results in a retinoblastoma protein defective in phosphorylation and oncoprotein binding. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6922–6926. doi: 10.1073/pnas.87.17.6922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Liu Y., Grandér D., Söderhäll S., Juliusson G., Gahrton G., Einhorn S. Retinoblastoma gene deletions in B-cell chronic lymphocytic leukemia. Genes Chromosomes Cancer. 1992 Apr;4(3):250–256. doi: 10.1002/gcc.2870040310. [DOI] [PubMed] [Google Scholar]
  14. Losada A. P., Wessman M., Tiainen M., Hopman A. H., Willard H. F., Solé F., Caballín M. R., Woessner S., Knuutila S. Trisomy 12 in chronic lymphocytic leukemia: an interphase cytogenetic study. Blood. 1991 Aug 1;78(3):775–779. [PubMed] [Google Scholar]
  15. Marshall C. J. Tumor suppressor genes. Cell. 1991 Jan 25;64(2):313–326. doi: 10.1016/0092-8674(91)90641-b. [DOI] [PubMed] [Google Scholar]
  16. Orita M., Suzuki Y., Sekiya T., Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics. 1989 Nov;5(4):874–879. doi: 10.1016/0888-7543(89)90129-8. [DOI] [PubMed] [Google Scholar]
  17. Shew J. Y., Chen P. L., Bookstein R., Lee E. Y., Lee W. H. Deletion of a splice donor site ablates expression of the following exon and produces an unphosphorylated RB protein unable to bind SV40 T antigen. Cell Growth Differ. 1990 Jan;1(1):17–25. [PubMed] [Google Scholar]
  18. Szekely L., Uzvolgyi E., Jiang W. Q., Durko M., Wiman K. G., Klein G., Sumegi J. Subcellular localization of the retinoblastoma protein. Cell Growth Differ. 1991 Jun;2(6):287–295. [PubMed] [Google Scholar]