Skip to main content
PMC home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1998;77(3):466–471. doi: 10.1038/bjc.1998.74

Reduced expression of deleted colorectal carcinoma (DCC) protein in established colon cancers.

T Goi 1, A Yamaguchi 1, G Nakagawara 1, T Urano 1, H Shiku 1, K Furukawa 1
PMCID: PMC2151282  PMID: 9472645

Abstract

Using a bacterial fusion protein, a deleted colorectal carcinoma (DCC)-specific monoclonal antibody (MAb) 127-22 was established. Although MAb 127-22 reacted with almost all normal tissues, it did not react or only weakly reacted with many cancer cell lines, including colonic cancer lines, in flow cytometry. In Western immunoblots, the MAb reacted with a single 190-kDa molecule in a myeloma line Ara-10 extract. This component was scarcely detected in colonic cancer cell lines. Immunoblots of samples from 25 pairs of colonic cancers and adjacent normal tissues and from five adenoma tissues revealed that all normal colonic and adenoma tissues significantly expressed the DCC protein, whereas colonic cancer tissues showed poor expression. These results indicate not only deletion of and lowered mRNA expression of the DCC gene, but also marked reduction of DCC protein occurred in colonic cancer tissues. In addition, colonic cancer patients with liver metastasis expressed significantly lower levels of DCC than those without, suggesting the prognostic value of DCC expression.

Full text

PDF

Images in this article

468Figure 1
on p.468
469Figure 3
on p.469
469Figure 4
on p.469
469Figure 5
on p.469

Selected References

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

  1. Baker S. J., Fearon E. R., Nigro J. M., Hamilton S. R., Preisinger A. C., Jessup J. M., vanTuinen P., Ledbetter D. H., Barker D. F., Nakamura Y. Chromosome 17 deletions and p53 gene mutations in colorectal carcinomas. Science. 1989 Apr 14;244(4901):217–221. doi: 10.1126/science.2649981. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Cho J. H., Noguchi M., Ochiai A., Hirohashi S. Loss of heterozygosity of multiple tumor suppressor genes in human gastric cancers by polymerase chain reaction. Lab Invest. 1996 Apr;74(4):835–841. [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Ekstrand B. C., Mansfield T. A., Bigner S. H., Fearon E. R. DCC expression is altered by multiple mechanisms in brain tumours. Oncogene. 1995 Dec 7;11(11):2393–2402. [PubMed] [Google Scholar]
  6. Fearon E. R., Cho K. R., Nigro J. M., Kern S. E., Simons J. W., Ruppert J. M., Hamilton S. R., Preisinger A. C., Thomas G., Kinzler K. W. Identification of a chromosome 18q gene that is altered in colorectal cancers. Science. 1990 Jan 5;247(4938):49–56. doi: 10.1126/science.2294591. [DOI] [PubMed] [Google Scholar]
  7. Fearon E. R., Ekstrand B. C., Hu G., Pierceall W. E., Reale M. A., Bigner S. H. Studies of the deleted in colorectal cancer gene in normal and neoplastic tissues. Cold Spring Harb Symp Quant Biol. 1994;59:637–643. doi: 10.1101/sqb.1994.059.01.073. [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. Hedrick L., Cho K. R., Boyd J., Risinger J., Vogelstein B. DCC: a tumor suppressor gene expressed on the cell surface. Cold Spring Harb Symp Quant Biol. 1992;57:345–351. doi: 10.1101/sqb.1992.057.01.039. [DOI] [PubMed] [Google Scholar]
  10. Hedrick L., Cho K. R., Fearon E. R., Wu T. C., Kinzler K. W., Vogelstein B. The DCC gene product in cellular differentiation and colorectal tumorigenesis. Genes Dev. 1994 May 15;8(10):1174–1183. doi: 10.1101/gad.8.10.1174. [DOI] [PubMed] [Google Scholar]
  11. Höhne M. W., Halatsch M. E., Kahl G. F., Weinel R. J. Frequent loss of expression of the potential tumor suppressor gene DCC in ductal pancreatic adenocarcinoma. Cancer Res. 1992 May 1;52(9):2616–2619. [PubMed] [Google Scholar]
  12. Iacopetta B., DiGrandi S., Dix B., Haig C., Soong R., House A. Loss of heterozygosity of tumour suppressor gene loci in human colorectal carcinoma. Eur J Cancer. 1994;30A(5):664–670. doi: 10.1016/0959-8049(94)90541-x. [DOI] [PubMed] [Google Scholar]
  13. Iino H., Fukayama M., Maeda Y., Koike M., Mori T., Takahashi T., Kikuchi-Yanoshita R., Miyaki M., Mizuno S., Watanabe S. Molecular genetics for clinical management of colorectal carcinoma. 17p, 18q, and 22q loss of heterozygosity and decreased DCC expression are correlated with the metastatic potential. Cancer. 1994 Mar 1;73(5):1324–1331. doi: 10.1002/1097-0142(19940301)73:5<1324::aid-cncr2820730503>3.0.co;2-w. [DOI] [PubMed] [Google Scholar]
  14. Inokuchi K., Miyake K., Takahashi H., Dan K., Nomura T. DCC protein expression in hematopoietic cell populations and its relation to leukemogenesis. J Clin Invest. 1996 Feb 1;97(3):852–857. doi: 10.1172/JCI118486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Itoh F., Hinoda Y., Ohe M., Ohe Y., Ban T., Endo T., Imai K., Yachi A. Decreased expression of DCC mRNA in human colorectal cancers. Int J Cancer. 1993 Jan 21;53(2):260–263. doi: 10.1002/ijc.2910530215. [DOI] [PubMed] [Google Scholar]
  16. Kashiwaba M., Tamura G., Ishida M. Frequent loss of heterozygosity at the deleted in colorectal carcinoma gene locus and its association with histologic phenotypes in breast carcinoma. Virchows Arch. 1995;426(5):441–446. doi: 10.1007/BF00193166. [DOI] [PubMed] [Google Scholar]
  17. Kataoka M., Okabayashi T., Orita K. Decreased expression of DCC mRNA in gastric and colorectal cancer. Surg Today. 1995;25(12):1001–1007. doi: 10.1007/BF00311682. [DOI] [PubMed] [Google Scholar]
  18. Kato M., Ito Y., Kobayashi S., Isono K. Detection of DCC and Ki-ras gene alterations in colorectal carcinoma tissue as prognostic markers for liver metastatic recurrence. Cancer. 1996 Apr 15;77(8 Suppl):1729–1735. doi: 10.1002/(SICI)1097-0142(19960415)77:8<1729::AID-CNCR47>3.0.CO;2-Z. [DOI] [PubMed] [Google Scholar]
  19. Kern S. E., Fearon E. R., Tersmette K. W., Enterline J. P., Leppert M., Nakamura Y., White R., Vogelstein B., Hamilton S. R. Clinical and pathological associations with allelic loss in colorectal carcinoma [corrected]. JAMA. 1989 Jun 2;261(21):3099–3103. doi: 10.1001/jama.261.21.3099. [DOI] [PubMed] [Google Scholar]
  20. Kikuchi-Yanoshita R., Konishi M., Fukunari H., Tanaka K., Miyaki M. Loss of expression of the DCC gene during progression of colorectal carcinomas in familial adenomatous polyposis and non-familial adenomatous polyposis patients. Cancer Res. 1992 Jul 1;52(13):3801–3803. [PubMed] [Google Scholar]
  21. Klingelhutz A. J., Hedrick L., Cho K. R., McDougall J. K. The DCC gene suppresses the malignant phenotype of transformed human epithelial cells. Oncogene. 1995 Apr 20;10(8):1581–1586. [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lawlor K. G., Narayanan R. Persistent expression of the tumor suppressor gene DCC is essential for neuronal differentiation. Cell Growth Differ. 1992 Sep;3(9):609–616. [PubMed] [Google Scholar]
  24. Miyake S., Nagai K., Yoshino K., Oto M., Endo M., Yuasa Y. Point mutations and allelic deletion of tumor suppressor gene DCC in human esophageal squamous cell carcinomas and their relation to metastasis. Cancer Res. 1994 Jun 1;54(11):3007–3010. [PubMed] [Google Scholar]
  25. Murty V. V., Li R. G., Houldsworth J., Bronson D. L., Reuter V. E., Bosl G. J., Chaganti R. S. Frequent allelic deletions and loss of expression characterize the DCC gene in male germ cell tumors. Oncogene. 1994 Nov;9(11):3227–3231. [PubMed] [Google Scholar]
  26. Nowell P. C. Mechanisms of tumor progression. Cancer Res. 1986 May;46(5):2203–2207. [PubMed] [Google Scholar]
  27. Ookawa K., Sakamoto M., Hirohashi S., Yoshida Y., Sugimura T., Terada M., Yokota J. Concordant p53 and DCC alterations and allelic losses on chromosomes 13q and 14q associated with liver metastases of colorectal carcinoma. Int J Cancer. 1993 Feb 1;53(3):382–387. doi: 10.1002/ijc.2910530307. [DOI] [PubMed] [Google Scholar]
  28. Sato Y., Mukai K., Watanabe S., Goto M., Shimosato Y. The AMeX method. A simplified technique of tissue processing and paraffin embedding with improved preservation of antigens for immunostaining. Am J Pathol. 1986 Dec;125(3):431–435. [PMC free article] [PubMed] [Google Scholar]
  29. Shibata D., Reale M. A., Lavin P., Silverman M., Fearon E. R., Steele G., Jr, Jessup J. M., Loda M., Summerhayes I. C. The DCC protein and prognosis in colorectal cancer. N Engl J Med. 1996 Dec 5;335(23):1727–1732. doi: 10.1056/NEJM199612053352303. [DOI] [PubMed] [Google Scholar]
  30. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  31. Thiagalingam S., Lengauer C., Leach F. S., Schutte M., Hahn S. A., Overhauser J., Willson J. K., Markowitz S., Hamilton S. R., Kern S. E. Evaluation of candidate tumour suppressor genes on chromosome 18 in colorectal cancers. Nat Genet. 1996 Jul;13(3):343–346. doi: 10.1038/ng0796-343. [DOI] [PubMed] [Google Scholar]
  32. Turley H., Pezzella F., Kocialkowski S., Comley M., Kaklamanis L., Fawcett J., Simmons D., Harris A. L., Gatter K. C. The distribution of the deleted in colon cancer (DCC) protein in human tissues. Cancer Res. 1995 Dec 1;55(23):5628–5631. [PubMed] [Google Scholar]
  33. Vogelstein B., Fearon E. R., Hamilton S. R., Kern S. E., Preisinger A. C., Leppert M., Nakamura Y., White R., Smits A. M., Bos J. L. Genetic alterations during colorectal-tumor development. N Engl J Med. 1988 Sep 1;319(9):525–532. doi: 10.1056/NEJM198809013190901. [DOI] [PubMed] [Google Scholar]
  34. Yamashiro S., Ruan S., Furukawa K., Tai T., Lloyd K. O., Shiku H., Furukawa K. Genetic and enzymatic basis for the differential expression of GM2 and GD2 gangliosides in human cancer cell lines. Cancer Res. 1993 Nov 15;53(22):5395–5400. [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES