According to the classical interpretation of Knudson's 'two-hit' hypothesis for tumorigenesis, the two 'hits' are independent mutation events, the end result of which is loss of a tumor suppressing function. Recently, it has been shown that the APC (adenomatous polyposis coli) gene does not entirely follow this model. Both the position and type of the second hit in familial adenomatous polyposis (FAP) polyps depend on the localization of the germline mutation. This non-random distribution of somatic hits has been interpreted as the result of selection for more advantageous mutations during tumor formation. However, the APC gene encodes for a multifunctional protein, and the exact cellular function upon which this selection is based is yet unknown. In this study, we have analyzed somatic APC point mutations and loss of heterozygosity (LOH) in 133 colorectal adenomas from six FAP patients. We observed that when germline mutations result in truncated proteins without any of the seven β-catenin downregulating 20-amino-acid repeats distributed in the central domain of APC, the majority of the corresponding somatic point mutations retain one or, less frequently, two of the same 20-amino-acid repeats. Conversely, when the germline mutation results in a truncated protein retaining one 20-amino-acid repeat, most second hits remove all 20-amino-acid repeats. The latter is frequently accomplished by allelic loss. Notably, and in contrast to previous observations, in a patient where the germline APC mutation retains two such repeats, the majority of the somatic hits are point mutations (and not LOH) located upstream and removing all of the 20-amino-acid repeats. These results indicate selection for APC genotypes that are likely to retain some activity in downregulating β-catenin signaling. We propose that this selection process is aimed at a specific degree of β-catenin signaling optimal for tumor formation, rather than at its constitutive activation by deletion of all the β-catenin downregulating motifs in APC.
|Number of pages
|Human Molecular Genetics
|Published - 15 Jun 2002
Bibliographical noteFunding Information:
We thank Cátia Marques for her assistance with sequencing of the mutations, Peter de Knijff for his assistance with the statistical analyses, and Menno Kielman and Carli Tops for their helpful suggestions. This project was supported by the Portuguese Ministry of Health(1999), and by grants from the Dutch Cancer Society (98-1652) to R.S. and C.B.