Method for improved diagnosis of dysplasias
The present invention relates to a method for improved diagnosis of dysplasias based on simultaneous detection of INK4a gene products and at least one marker for cell proliferation. Particularly the present invention provides a method for discriminating dysplastic cells over-expressing INK4a gene products from cells over-expressing INK4a gene products without being dysplastic by detection of a marker suitable for characterizing the proliferation properties of the respective cell. The characterization of the proliferation properties may comprise the detection of a marker or a set of markers characteristic for active cell proliferation and/or a marker or a set of markers characteristic for retarded or ceased cell proliferation. The method presented herein thus enables for a specific diagnosis of dysplasias in histological and cytological specimens.
1. A method of discriminating cervical dysplastic lesions from metaplastic lesions in a subject, the method comprising:
(a) measuring an expression level of a high-risk HPV-associated nucleic acid in a sample collected from a cervical lesion in a subject;
(b) measuring an expression level of at least one human protein marker selected from the group consisting of p14ARF and p16 INK4a in the sample;
(c) measuring an expression level of at least one human proliferation protein marker selected from a group consisting of MCM2, MCM5, CDC6, PCNA, and Ki67 in the sample;
(d) determining that the lesion is dysplastic if there is a detectable level of the high-risk HPV-associated nucleic acid in the sample and there is at least one cell in the sample simultaneously expressing the protein marker of (b) and the proliferation protein marker of (c); and
(e) determining that the lesion is metaplastic if there is a detectable level of the high-risk HPV-associated nucleic acid in the sample and if no cell in the sample co-expresses the protein marker of (b) and the proliferation protein marker of (c); thereby discriminating between dysplastic lesions from metaplastic lesions.
2. The method according to claim 1 , wherein the protein marker of (b) is P16 INK4a .
3. The method according to claim 1 , wherein the proliferation protein marker of (c) is MCM2 or MCM5.