P53, MAPK, topoisomerase II alpha and Ki67 immunohistochemical expression and KRAS/BRAF mutation in ovarian serous carcinomas

Diagn Pathol. 2013 Feb 6:8:21. doi: 10.1186/1746-1596-8-21.

Abstract

Background: We investigated the immunohistochemical expression of p53, MAPK, topoisomerase II alpha (topoII alpha) and Ki67 in ovarian serous carcinomas (OSCs) along with mutational analysis for KRAS and BRAF.

Methods: Eighty one cases of OSCs were reviewed and examined immunohistochemically using antibodies against p53, MAPK, topoII alpha and Ki67. Staining was evaluated as a percentage of immunopositive cells with cut-off levels at 10% for p53 and topoII alpha, and 5% for MAPK. The Ki67 immunoexpression was assessed by means of Olympus Image Analysis System as a percentage of immunopositive cells in 1000 tumor cells. KRAS and BRAF mutational analysis was performed on 73 available microdissected samples.

Results: Of 81 cases of OSCs 13.6% were of low-grade and 86.4% were of high-grade morphology. In the high-grade group there was a significantly higher immunoexpression of p53 (P < 0.001) and topoII alpha (P = 0.001), with Ki67 median 56.5 vs. 19 in low-grade group (P < 0.001). The difference in immunoexpression of active MAPK between low- and high-grade group was also significant (P = 0.003). MAPK positive immunostaining was detected in 63.6% of low-grade vs. 17.1% of high-grade OSCs. The frequency of KRAS mutation was significantly higher in low-grade as compared to high-grade group (P = 0.006). None of the samples had BRAF mutation. In addition, we detected positive MAPK immunoexpression in 13/59 samples with wild-type KRAS, suggesting that activation of MAPK pathway is not ultimately related either to KRAS or BRAF mutation. Seven morphologically high-grade samples (11.7%) showed both KRAS mutation and p53 immunopositivity.

Conclusions: Although this study is limited by its humble number of low-grade samples, our data fit the proposed dualistic pathway of ovarian carcinogenesis. Mutational analysis for KRAS and BRAF discloses some possible interactions between different tumorigenic pathways of low- and high-grade carcinomas. Immunohistochemical staining for MAPK was not sufficiently sensitive, nor specific, to precisely predict the KRAS mutation. However, it appears to be quite reliable in ruling out a KRAS mutation if the staining is negative.

Virtual slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/9283563368804632.

MeSH terms

  • Adult
  • Aged
  • Antigens, Neoplasm / analysis*
  • Biomarkers, Tumor* / analysis
  • Biomarkers, Tumor* / genetics
  • Carcinoma* / chemistry
  • Carcinoma* / genetics
  • Carcinoma* / pathology
  • Chi-Square Distribution
  • DNA Mutational Analysis
  • DNA Topoisomerases, Type II / analysis*
  • DNA-Binding Proteins / analysis*
  • Female
  • Humans
  • Immunohistochemistry*
  • Ki-67 Antigen / analysis*
  • Microdissection
  • Middle Aged
  • Mitogen-Activated Protein Kinases / analysis*
  • Mutation*
  • Neoplasm Grading
  • Neoplasms, Cystic, Mucinous, and Serous* / chemistry
  • Neoplasms, Cystic, Mucinous, and Serous* / genetics
  • Neoplasms, Cystic, Mucinous, and Serous* / pathology
  • Ovarian Neoplasms* / chemistry
  • Ovarian Neoplasms* / genetics
  • Ovarian Neoplasms* / pathology
  • Polymerase Chain Reaction
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins B-raf / genetics*
  • Proto-Oncogene Proteins p21(ras)
  • Tumor Suppressor Protein p53 / analysis*
  • ras Proteins / genetics*

Substances

  • Antigens, Neoplasm
  • Biomarkers, Tumor
  • DNA-Binding Proteins
  • KRAS protein, human
  • Ki-67 Antigen
  • Proto-Oncogene Proteins
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • Mitogen-Activated Protein Kinases
  • Proto-Oncogene Proteins p21(ras)
  • ras Proteins
  • DNA Topoisomerases, Type II