Skip to main content
NCBI home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1996 Jul;149(1):237–245.

Intratumor heterogeneity of K-ras2 mutations in colorectal adenocarcinomas: association with degree of DNA aneuploidy.

W Giaretti 1, R Monaco 1, N Pujic 1, A Rapallo 1, S Nigro 1, E Geido 1
PMCID: PMC1865212  PMID: 8686748

Abstract

Detailed information about intratumor K-ras2 mutations in colorectal adenocarcinomas and a possible association with DNA content heterogeneity is still lacking. DNA diploid and aneuploid subclones, detected among multiple histologically selected primary sectors (57 superficial and 40 deep) and 9 lymph node metastases, were flow cytometrically sorted and separately submitted to codons 12-13 K-ras2 mutation spectrum analysis. DNA aneuploidy was absent among 20 near and 20 distant mucosa sites and present in 7/9 lymph node metastases and in 17/19 primary tumors (90%). Primary intratumor DNA multiclonality was approximately 50%. Degree of DNA aneuploidy (DNA Index) distribution was nonrandom and showed peaks at approximate mean DNA Index values 1.2, 1.5, and 1.8. K-ras2 mutations were detected in 0/20 mucosa cases, in 2/9 lymph node metastases, and in 9/19 adenocarcinomas (47%). No more than one mutation type per tumor was detected. Intratumor distribution of K-ras2 mutations was homogeneous in 6 and heterogeneous in 3 cases. Homogeneous distribution was associated with DNA near-diploid aneuploidy. K-ras2 mutations were strongly associated with DNA Index in the near-diploid region (83%) and almost absent (5%) among DNA near-triploid subclones (P = 0.0001). K-ras2 mutation intratumor heterogeneity indicates that sampling of the tumor may be a critical step and suggests that K-ras2 activation may be a late event in a subgroup of tumors. Our data also suggest the existence of an early process of the colorectal carcinogenesis that favors both K-ras2 mutations and DNA near-diploid aneuploidy. Onset of DNA near-triploid subclones appears, instead, to be independent from K-ras2 activation.

Full text

PDF
237

Images in this article

241Figure 1
on p.241

Selected References

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

  1. ASTLER V. B., COLLER F. A. The prognostic significance of direct extension of carcinoma of the colon and rectum. Ann Surg. 1954 Jun;139(6):846–852. doi: 10.1097/00000658-195406000-00015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  3. Bauer K. D., Bagwell C. B., Giaretti W., Melamed M., Zarbo R. J., Witzig T. E., Rabinovitch P. S. Consensus review of the clinical utility of DNA flow cytometry in colorectal cancer. Cytometry. 1993;14(5):486–491. doi: 10.1002/cyto.990140506. [DOI] [PubMed] [Google Scholar]
  4. Benhattar J., Losi L., Chaubert P., Givel J. C., Costa J. Prognostic significance of K-ras mutations in colorectal carcinoma. Gastroenterology. 1993 Apr;104(4):1044–1048. doi: 10.1016/0016-5085(93)90272-e. [DOI] [PubMed] [Google Scholar]
  5. Bishop J. M. The molecular genetics of cancer. Science. 1987 Jan 16;235(4786):305–311. doi: 10.1126/science.3541204. [DOI] [PubMed] [Google Scholar]
  6. Bos J. L., Fearon E. R., Hamilton S. R., Verlaan-de Vries M., van Boom J. H., van der Eb A. J., Vogelstein B. Prevalence of ras gene mutations in human colorectal cancers. 1987 May 28-Jun 3Nature. 327(6120):293–297. doi: 10.1038/327293a0. [DOI] [PubMed] [Google Scholar]
  7. Bos J. L. p21ras: an oncoprotein functioning in growth factor-induced signal transduction. Eur J Cancer. 1995 Jul-Aug;31A(7-8):1051–1054. doi: 10.1016/0959-8049(95)00168-i. [DOI] [PubMed] [Google Scholar]
  8. Bos J. L. ras oncogenes in human cancer: a review. Cancer Res. 1989 Sep 1;49(17):4682–4689. [PubMed] [Google Scholar]
  9. Breivik J., Meling G. I., Spurkland A., Rognum T. O., Gaudernack G. K-ras mutation in colorectal cancer: relations to patient age, sex and tumour location. Br J Cancer. 1994 Feb;69(2):367–371. doi: 10.1038/bjc.1994.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Brown K., Buchmann A., Balmain A. Carcinogen-induced mutations in the mouse c-Ha-ras gene provide evidence of multiple pathways for tumor progression. Proc Natl Acad Sci U S A. 1990 Jan;87(2):538–542. doi: 10.1073/pnas.87.2.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Burmer G. C., Loeb L. A. Mutations in the KRAS2 oncogene during progressive stages of human colon carcinoma. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2403–2407. doi: 10.1073/pnas.86.7.2403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cairns J. The origin of human cancers. Nature. 1981 Jan 29;289(5796):353–357. doi: 10.1038/289353a0. [DOI] [PubMed] [Google Scholar]
  13. Cannon-Albright L. A., Skolnick M. H., Bishop D. T., Lee R. G., Burt R. W. Common inheritance of susceptibility to colonic adenomatous polyps and associated colorectal cancers. N Engl J Med. 1988 Sep 1;319(9):533–537. doi: 10.1056/NEJM198809013190902. [DOI] [PubMed] [Google Scholar]
  14. Capella G., Cronauer-Mitra S., Pienado M. A., Perucho M. Frequency and spectrum of mutations at codons 12 and 13 of the c-K-ras gene in human tumors. Environ Health Perspect. 1991 Jun;93:125–131. doi: 10.1289/ehp.9193125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fearon E. R. K-ras gene mutation as a pathogenetic and diagnostic marker in human cancer. J Natl Cancer Inst. 1993 Dec 15;85(24):1978–1980. doi: 10.1093/jnci/85.24.1978. [DOI] [PubMed] [Google Scholar]
  16. Fearon E. R., Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990 Jun 1;61(5):759–767. doi: 10.1016/0092-8674(90)90186-i. [DOI] [PubMed] [Google Scholar]
  17. Finkelstein S. D., Sayegh R., Christensen S., Swalsky P. A. Genotypic classification of colorectal adenocarcinoma. Biologic behavior correlates with K-ras-2 mutation type. Cancer. 1993 Jun 15;71(12):3827–3838. doi: 10.1002/1097-0142(19930615)71:12<3827::aid-cncr2820711207>3.0.co;2-n. [DOI] [PubMed] [Google Scholar]
  18. Fischbach W., Zidianakis Z., Lüke G., Kirchner T., Mössner J. DNA mapping of colorectal neoplasms: a flow cytometric study of DNA abnormalities and proliferation. Gastroenterology. 1993 Oct;105(4):1126–1133. doi: 10.1016/0016-5085(93)90958-f. [DOI] [PubMed] [Google Scholar]
  19. Fishel R., Lescoe M. K., Rao M. R., Copeland N. G., Jenkins N. A., Garber J., Kane M., Kolodner R. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell. 1993 Dec 3;75(5):1027–1038. doi: 10.1016/0092-8674(93)90546-3. [DOI] [PubMed] [Google Scholar]
  20. Forrester K., Almoguera C., Han K., Grizzle W. E., Perucho M. Detection of high incidence of K-ras oncogenes during human colon tumorigenesis. 1987 May 28-Jun 3Nature. 327(6120):298–303. doi: 10.1038/327298a0. [DOI] [PubMed] [Google Scholar]
  21. Giaretti W. A model of DNA aneuploidization and evolution in colorectal cancer. Lab Invest. 1994 Dec;71(6):904–910. [PubMed] [Google Scholar]
  22. Giaretti W., Pujic N., Rapallo A., Nigro S., Di Vinci A., Geido E., Risio M. K-ras-2 G-C and G-T transversions correlate with DNA aneuploidy in colorectal adenomas. Gastroenterology. 1995 Apr;108(4):1040–1047. doi: 10.1016/0016-5085(95)90201-5. [DOI] [PubMed] [Google Scholar]
  23. Giaretti W., Santi L. Tumor progression by DNA flow cytometry in human colorectal cancer. Int J Cancer. 1990 Apr 15;45(4):597–603. doi: 10.1002/ijc.2910450404. [DOI] [PubMed] [Google Scholar]
  24. Giaretti W., Sciallero S., Bruno S., Geido E., Aste H., Di Vinci A. DNA flow cytometry of endoscopically examined colorectal adenomas and adenocarcinomas. Cytometry. 1988 May;9(3):238–244. doi: 10.1002/cyto.990090309. [DOI] [PubMed] [Google Scholar]
  25. Hagag N., Diamond L., Palermo R., Lyubsky S. High expression of ras p21 correlates with increased rate of abnormal mitosis in NIH3T3 cells. Oncogene. 1990 Oct;5(10):1481–1489. [PubMed] [Google Scholar]
  26. Hartwell L. H., Kastan M. B. Cell cycle control and cancer. Science. 1994 Dec 16;266(5192):1821–1828. doi: 10.1126/science.7997877. [DOI] [PubMed] [Google Scholar]
  27. Hartwell L. Defects in a cell cycle checkpoint may be responsible for the genomic instability of cancer cells. Cell. 1992 Nov 13;71(4):543–546. doi: 10.1016/0092-8674(92)90586-2. [DOI] [PubMed] [Google Scholar]
  28. Hiddemann W., Von Bassewitz D. B., Kleinemeier H. J., Schulte-Brochterbeck E., Hauss J., Lingemann B., Büchner T., Grundmann E. DNA stemline heterogeneity in colorectal cancer. Cancer. 1986 Jul 15;58(2):258–263. doi: 10.1002/1097-0142(19860715)58:2<258::aid-cncr2820580210>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  29. Kaufmann W. K. Cell cycle checkpoints and DNA repair preserve the stability of the human genome. Cancer Metastasis Rev. 1995 Mar;14(1):31–41. doi: 10.1007/BF00690209. [DOI] [PubMed] [Google Scholar]
  30. Krengel U., Schlichting I., Scherer A., Schumann R., Frech M., John J., Kabsch W., Pai E. F., Wittinghofer A. Three-dimensional structures of H-ras p21 mutants: molecular basis for their inability to function as signal switch molecules. Cell. 1990 Aug 10;62(3):539–548. doi: 10.1016/0092-8674(90)90018-a. [DOI] [PubMed] [Google Scholar]
  31. Laurent-Puig P., Olschwang S., Delattre O., Remvikos Y., Asselain B., Melot T., Validire P., Muleris M., Girodet J., Salmon R. J. Survival and acquired genetic alterations in colorectal cancer. Gastroenterology. 1992 Apr;102(4 Pt 1):1136–1141. [PubMed] [Google Scholar]
  32. Loeb L. A. Endogenous carcinogenesis: molecular oncology into the twenty-first century--presidential address. Cancer Res. 1989 Oct 15;49(20):5489–5496. [PubMed] [Google Scholar]
  33. McLellan E. A., Owen R. A., Stepniewska K. A., Sheffield J. P., Lemoine N. R. High frequency of K-ras mutations in sporadic colorectal adenomas. Gut. 1993 Mar;34(3):392–396. doi: 10.1136/gut.34.3.392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Moerkerk P., Arends J. W., van Driel M., de Bruïne A., de Goeij A., ten Kate J. Type and number of Ki-ras point mutations relate to stage of human colorectal cancer. Cancer Res. 1994 Jul 1;54(13):3376–3378. [PubMed] [Google Scholar]
  35. Muto T., Bussey H. J., Morson B. C. The evolution of cancer of the colon and rectum. Cancer. 1975 Dec;36(6):2251–2270. doi: 10.1002/cncr.2820360944. [DOI] [PubMed] [Google Scholar]
  36. Nicolaides N. C., Papadopoulos N., Liu B., Wei Y. F., Carter K. C., Ruben S. M., Rosen C. A., Haseltine W. A., Fleischmann R. D., Fraser C. M. Mutations of two PMS homologues in hereditary nonpolyposis colon cancer. Nature. 1994 Sep 1;371(6492):75–80. doi: 10.1038/371075a0. [DOI] [PubMed] [Google Scholar]
  37. Nowell P. C. Cancer, chromosomes, and genes. Lab Invest. 1992 Apr;66(4):407–417. [PubMed] [Google Scholar]
  38. Nowell P. C. Molecular events in tumor development. N Engl J Med. 1988 Sep 1;319(9):575–577. doi: 10.1056/NEJM198809013190908. [DOI] [PubMed] [Google Scholar]
  39. Nowell P. C. The clonal evolution of tumor cell populations. Science. 1976 Oct 1;194(4260):23–28. doi: 10.1126/science.959840. [DOI] [PubMed] [Google Scholar]
  40. Pretlow T. P. Aberrant crypt foci and K-ras mutations: earliest recognized players or innocent bystanders in colon carcinogenesis? Gastroenterology. 1995 Feb;108(2):600–603. doi: 10.1016/0016-5085(95)90092-6. [DOI] [PubMed] [Google Scholar]
  41. Rubio C. A. Atypical mitoses in colorectal adenomas. Pathol Res Pract. 1991 May;187(4):508–513. doi: 10.1016/S0344-0338(11)80015-4. [DOI] [PubMed] [Google Scholar]
  42. Santos E., Martin-Zanca D., Reddy E. P., Pierotti M. A., Della Porta G., Barbacid M. Malignant activation of a K-ras oncogene in lung carcinoma but not in normal tissue of the same patient. Science. 1984 Feb 17;223(4637):661–664. doi: 10.1126/science.6695174. [DOI] [PubMed] [Google Scholar]
  43. Sciallero S., Bruno S., Di Vinci A., Geido E., Aste H., Giaretti W. Flow cytometric DNA ploidy in colorectal adenomas and family history of colorectal cancer. Cancer. 1988 Jan 1;61(1):114–120. doi: 10.1002/1097-0142(19880101)61:1<114::aid-cncr2820610120>3.0.co;2-i. [DOI] [PubMed] [Google Scholar]
  44. Solomon E., Voss R., Hall V., Bodmer W. F., Jass J. R., Jeffreys A. J., Lucibello F. C., Patel I., Rider S. H. Chromosome 5 allele loss in human colorectal carcinomas. Nature. 1987 Aug 13;328(6131):616–619. doi: 10.1038/328616a0. [DOI] [PubMed] [Google Scholar]
  45. Suchy B., Zietz C., Rabes H. M. K-ras point mutations in human colorectal carcinomas: relation to aneuploidy and metastasis. Int J Cancer. 1992 Aug 19;52(1):30–33. doi: 10.1002/ijc.2910520107. [DOI] [PubMed] [Google Scholar]
  46. Sutherland G. R., Simmers R. N. No statistical association between common fragile sites and nonrandom chromosome breakpoints in cancer cells. Cancer Genet Cytogenet. 1988 Mar;31(1):9–15. doi: 10.1016/0165-4608(88)90004-0. [DOI] [PubMed] [Google Scholar]
  47. Topal M. D. DNA repair, oncogenes and carcinogenesis. Carcinogenesis. 1988 May;9(5):691–696. doi: 10.1093/carcin/9.5.691. [DOI] [PubMed] [Google Scholar]
  48. Urosević N., Krtolica K., Skaro-Milić A., Knezević-Usaj S., Dujić A. Prevalence of G-to-T transversions among K-ras oncogene mutations in human colorectal tumors in Yugoslavia. Int J Cancer. 1993 May 8;54(2):249–254. doi: 10.1002/ijc.2910540215. [DOI] [PubMed] [Google Scholar]
  49. Verlaan-de Vries M., Bogaard M. E., van den Elst H., van Boom J. H., van der Eb A. J., Bos J. L. A dot-blot screening procedure for mutated ras oncogenes using synthetic oligodeoxynucleotides. Gene. 1986;50(1-3):313–320. doi: 10.1016/0378-1119(86)90335-5. [DOI] [PubMed] [Google Scholar]
  50. 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]
  51. Vogelstein B., Fearon E. R., Kern S. E., Hamilton S. R., Preisinger A. C., Nakamura Y., White R. Allelotype of colorectal carcinomas. Science. 1989 Apr 14;244(4901):207–211. doi: 10.1126/science.2565047. [DOI] [PubMed] [Google Scholar]
  52. Volpe J. P. Genetic instability of cancer. Why a metastatic tumor is unstable and a benign tumor is stable. Cancer Genet Cytogenet. 1988 Aug;34(1):125–134. doi: 10.1016/0165-4608(88)90179-3. [DOI] [PubMed] [Google Scholar]
  53. Vousden K. H., Bos J. L., Marshall C. J., Phillips D. H. Mutations activating human c-Ha-ras1 protooncogene (HRAS1) induced by chemical carcinogens and depurination. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1222–1226. doi: 10.1073/pnas.83.5.1222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Weinberg R. A. Oncogenes, antioncogenes, and the molecular bases of multistep carcinogenesis. Cancer Res. 1989 Jul 15;49(14):3713–3721. [PubMed] [Google Scholar]
  55. Yamashita N., Minamoto T., Ochiai A., Onda M., Esumi H. Frequent and characteristic K-ras activation and absence of p53 protein accumulation in aberrant crypt foci of the colon. Gastroenterology. 1995 Feb;108(2):434–440. doi: 10.1016/0016-5085(95)90071-3. [DOI] [PubMed] [Google Scholar]
  56. Yunis J. J., Soreng A. L. Constitutive fragile sites and cancer. Science. 1984 Dec 7;226(4679):1199–1204. doi: 10.1126/science.6239375. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES