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In situ localization of S-phase-specific histone (H3) mRNA in Bowen's disease
M Murakami1, Y Mizoguchi, Y Horibe
1Department of Pathology, Fujita Health University, School of Medicine, Aichi, Japan.
Abstract:
PCNA and Ki-67 immunohistochemistry has been used to assess cell proliferation in place of tritiated thymidine or BrdU labeling of S-phase cells. Recently, it has been possible to reliably demonstrate histone H3 mRNA by in situ hybridization in formalin-fixed and paraffin-embedded tissue sections. We have compared this new proliferation marker with Ki-67 and PCNA with regard to distribution of positive cells and labeling indices (LI%) for 22 cases of Bowen's disease. In normal skin, Ki-67-IHC positive cells and histone mRNA positive cells were observed in the basal and suprabasal layers of the epidermis. In Bowen's disease, positive cells with each marker were more frequent in upper neoplastic epidermis than in suprabasal layers, and the average LI%s were markedly elevated with all markers, the scores decreasing in the following order: PCNA-IHC, Ki-67-IHC and H3mRNA-ISH. However, the results of double staining demonstrated that S-phase cells do not necessarily show exactly the same distributions as with PCNA and Ki-67-IHC labeling. H3mRNA-ISH showed three different degrees of reaction with significantly different LI%s, whereas PCNA and Ki-67 LI% did not vary essentially in the same areas. These results strongly suggest that Bowen's disease, which is well known as a low-grade neoplastic state with malignant potential, also demonstrates clear intratumoral heterogeneity of S-phase cells using the H3mRNA-ISH method.
Insights
Histone H3 mRNA in situ hybridization (H3mRNA-ISH) offers a new way to study cell proliferation. This method revealed significant intratumoral heterogeneity of S-phase cells in Bowen's disease, unlike traditional markers.
Area of Science:
- Dermatopathology
- Molecular Biology
- Oncology
Background:
- Proliferation markers like PCNA and Ki-67 are used in immunohistochemistry (IHC).
- Histone H3 mRNA in situ hybridization (H3mRNA-ISH) is a novel method for detecting cell proliferation in fixed tissues.
- Bowen's disease is a neoplastic skin condition with malignant potential.
Purpose of the Study:
- To compare H3mRNA-ISH with PCNA-IHC and Ki-67-IHC for assessing cell proliferation in Bowen's disease.
- To evaluate the distribution and labeling indices (LI%) of positive cells using these markers.
- To investigate intratumoral heterogeneity of S-phase cells in Bowen's disease.
Main Methods:
- Comparison of H3mRNA-ISH with PCNA-IHC and Ki-67-IHC in 22 cases of Bowen's disease.
- Analysis of cell distribution and labeling indices (LI%) for each marker.
- Double staining techniques to assess co-localization of markers.
Main Results:
- All markers showed elevated LI% in Bowen's disease compared to normal skin.
- LI% decreased in the order: PCNA-IHC > Ki-67-IHC > H3mRNA-ISH.
- H3mRNA-ISH revealed distinct patterns and LI% variations, indicating intratumoral heterogeneity not fully captured by PCNA and Ki-67.
Conclusions:
- H3mRNA-ISH is a valuable tool for detecting proliferation and intratumoral heterogeneity in Bowen's disease.
- Bowen's disease exhibits significant intratumoral heterogeneity of S-phase cells, as demonstrated by H3mRNA-ISH.
- This heterogeneity may have implications for understanding the neoplastic state and malignant potential of Bowen's disease.