Objective : Carcinosarcoma of the female genital organs is an uncommon but clinically highly aggressive neoplasm with biphasic histology of carcinomatous and sarcomatous components. In previous studies, we have found frequent loss of heterozygosity (LOH) at 9p21 (41.2%) the loci of the tumor suppressor gene
p16, in carcinosarcoma cases. Alteration of
p16 is known to be one of the most frequent genetic abnormalities in human neoplasias.
p16 is frequently inactivated by methylation of its CpG islands, and much less frequently by mutations or homozygous deletions. In the present study, we investigated whether abnormalities of
p16 are involved in the evolution of carcinosarcomas.
Methods : Eighteen cases of gynecological carcinosarcomas were submitted for genetic studies. DNA was prepared from microdissected paraffin sections. Mutation of
p16 was examined by the single-strand conformational polymorphism (SSCP) technique. Methylation status of
p16 was examined by methylation specific PCR (MSP) and methylation-sensitive restriction enzyme digest PCR (MSREDP). The methylation levels of
p16 in carcinomatous and sarcomatous foci were quantified by the combined bisulfite-restriction analysis (COBRA) technique.
Results : There were mutations detected in any carcinosarcoma cases. Four of 17 cases showed methylation in the
p16 promoter region, four of 17 showed methylation in exon 1, and 15 of 17 showed methylation in exon2. COBRA analysis of both the carcinomatous and sarcomatous components showed almost equal levels of methylation ranging from 27 to 33%. However, the methylation level of microdissected chondrosarcomatous elements was about 20%, which was approximately 10% lower than that of the other components.
Conclusions : Hypermethylation of s
p16 exon 2 but not promotor or exon 1 is a frequent abnormality in carcinosarcomas. Exon 2 methylation has been reported in some advanced cancers and in a few different types of cancers. Thus, 9p21 LOH together with methylation of
p16 exon 2 may be a unique mechanism for the two-hit inactivation of
p16in carcinosarcomas. Additionally, different levels of methylation in the neoplastic foci may affect the histological diversity of carcinosarcoma.
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